Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Priority
Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION. —The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claim 4 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Regarding Claim 4, the recitation “a proportion of organic residues in the sample is quantitatively determined before the image is generated and no detection signal is output if the proportion is below a predetermined threshold” renders the claim unclear and indefinite. Claim 4 depends from claim 1. Claim 1 previously recites “outputting a detection signal when there is detection in the image of one or more characteristic emission wavelengths of any of said one or more of said fluorescent markers that have fluorescently labeled any of the biofilm components present in the sample.” It is unclear as to whether the detection signal of claim 4 is the same detection signal as claim 1. It is unclear because claim 1 specifies a detection signal being generated when there is detection in the image but in claim 4, there is no detection signal output based on a determination performed before the image is generated. Further clarification is requested.
Claim Interpretation
Direct emission is being interpreted to mean that the fluorescent marker itself emits light in a certain wavelength range after excitation. Indirect emission means that the fluorescent marker interacts with another substance so that this other substance emits light in a certain wavelength range due to this interaction.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
Claims 1, 2, 8 are rejected under 35 U.S.C. 103 as being obvious over MEYN (DE 102017005695) in further view of MARTINO et al. (Assessing chemical cleaning of nanofiltration membranes in a drinking water production plant: a combination of chemical composition analysis and fluorescence microscopy) citations made with respect to the copy provided with IDS dated 02 June 2023.
With respect to claim 1, MEYN teaches of a method detecting a biofilm in a closed plant (detecting biofilms in production plants (1) for food and / or beverage production when the plant is emptied (Page 1 paragraph starting with “The invention relates”)), said method comprising:
introducing a rinsing medium into the plant when emptied (Page 2 paragraph starting with “Since the production” and Page 3 paragraph starting with “According to a practical”) , then admixing an enzymatic solution that detaches the biofilm and circulating a mixture therefrom in the plant (Page 4, paragraph starting with “That in the bypass cycle”), circulating the mixture therefrom is circulated (4) in the plant (Page 4, paragraph starting with “That in the bypass cycle”), taking a sample of the circulated mixture from the plant (Page 4, paragraph starting with “That in the bypass cycle”). MEYN also teaches measuring the total organic carbon as an indicator of biofilm presence (Page 5, paragraph starting with “If the comparison” and Page 4, paragraph starting with “According to an”) in interfaces of the plant such as a heat exchanger due the area being a flow-poor area and biofilms being encapsulated by a surface layer that is resistant to hot water or hot steam (Page 5, paragraph starting with “A special challenge”). MEYN does not teach then fluorescently labeling the sample with one or more fluorescent markers that each specifically bind to a respective biofilm component; and generating a photometric image of the sample; and then outputting a detection signal when there is detection in the image of one or more characteristic emission wavelengths of any of said one or more of said fluorescent markers that have fluorescently labeled any of the biofilm components present in the sample. MARTINO et al. is used to remedy this; MARTINO et al. teaches the analysis of fouling deposits removed from nanofiltration membranes to evaluate the cleaning process of foulants that may be present in drinking water plants (Page 220, paragraph starting with “The primary objective”). More specifically, MARTINO et al. teaches then fluorescently labeling the sample with one or more fluorescent markers that each specifically bind to a respective biofilm component plants (Page 220, paragraph starting with “Samples of the fouled membranes”). MARTINO et al. also teaches generating a photometric image of the sample (Figure 1 and Figure 4); and then outputting a detection signal when there is detection in the image of one or more characteristic emission wavelengths of any of said one or more of said fluorescent markers that have fluorescently labeled any of the biofilm components present in the sample (Figure 3).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to modify the detection of biofilm from an industrial plant by measuring the total organic carbon to instead use fluorescent markers that specifically bind to the respective biofilm removed from the plant, generate a photometric image of the sample; and then outputting a detection signal when there is detection in the image of one or more characteristic emission wavelengths of any of said one or more of said fluorescent markers that have fluorescently labeled any of the biofilm components present in the sample as taught by MARTINO et al because both MEYN and MARTINO et al focus on the detection of a biofilm in from a production plant, furthermore MARTINO et al. teaches using fluorescent labels that specifically bind to the biofilm components instead of detecting total organic carbon within the plant system, and this involves using a known method to substitute the MEYN reference to improve specificity in detection of biofilm as discussed in MPEP 2143(I)(B).
With respect to claim 2, Modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 1). Modified MEYN teaches wherein the image of the sample is generated using fluorescence microscopy (MARTINO et al, Figure 4).
With respect to claim 8, Modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 1.
MEYN teaches wherein the sample is a filter cake (surface layer of the biofilm) by an external rinsing cycle, into which the production plant or production plant part is investigated by being fluidly coupled to a measuring circuit (Page 2, paragraph starting with “ A device for carrying”). MEYN teaches particularly a heat exchanger and flow-poor areas of at least a part of the circulated mixture (Page 6, paragraph starting with “Instead of the entire” and Page 7, paragraph starting with “compared to the flow”). A rinsing cycle was introduced into the production plant/plant part coupled circuit as the first step of detecting the biofilm (Page 7, paragraph starting with “In the first process”) from the production plant/ production part.
However, MEYN does not teach specifically filter cake obtained by filtration.
MARTINO et al. is used to remedy this as MARTINO et al. teaches a cleaning in place process for nanofiltration units that are used in drinking water plants (Page 220, paragraph starting with “Nanofiltration membranes”). More specifically, MARTINO et al. teaches identifying the major components of fouling material from a filtration membrane surface (Page 220, paragraph starting with “The primary objective”). MARTINO et al. teaches foulants of several membrane sheets that were scraped for cake removal (Page 220, paragraph starting with “ Foulants of several membrane”). MARTINO et al. further teaches that the nanofiltration membranes were removed from a drinking water plant that was in operation for 6 years prior detection, cleaning and analysis of the foulant components (Page 220, paragraph starting with “Nanofiltration (NF) membranes”).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to modify the removal of biofilm surface layer in a heat exchanger and/or flow dead spaces of a food and beverage plant by MEYN to instead removing a cake foulant from membranes in a nanofiltration unit in a drinking water plant after filtration as taught by MARTINO et al. because both MEYN and MARTINO et al focus on removing and detecting biofilm from plant parts specifically flow restricted areas of the production plant, MEYN teaches detecting biofilm that is encapsulated by a surface layer that is removed from a food and beverage heat exchanger and/or flow dead spaces instead of removing cake from a nanofiltration unit of drinking plant membrane that were obtained after operation, and this involves simple substitution of the production plant to obtain predictable results as discussed in MPEP 2143(I)(B).
Claims 3, 5, 12, 14, are rejected under 35 U.S.C. 103 as being obvious over MEYN (DE 102017005695) in view of MARTINO (Assessing chemical cleaning of nanofiltration membranes in a drinking water production plant: a combination of chemical composition analysis and fluorescence microscopy) as applied to claims 1 and 2 above and further view of BOYETTE et al. (US 20100112630 A1), citation made with respect to the copy provided with IDS dated 02 June 2023.
With respect to claim 3, Modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 1.
Modified MEYN further teaches admixing an aqueous rinsing medium and enzymatic cleaner and the mixture circulating in the plant as specified previously (see above in claim 1) (MEYN, Page 4, paragraph starting with “That in the bypass cycle”).
Modified MEYN did not specify the rinsing medium is tempered to 40-50 degrees before admixing the enzymatic solution.
BOYETTE et al. is used to remedy this, as BOYETTE et al. teaches a method for measuring the total microbiological content of an aqueous medium with the released intracellular content of the microbiological matter from an industrial plant ([0006] and [0019]). BOYETTE et al. specifically teaches BOYETTE also teaches removing a portion off the aqueous medium as a testing sample for biofilm detection [0024]. BOYETTE et al. specifically teaches the temperature of the aqueous medium being 40-100 degrees before removing the sample for microbiological matter detection (0047). BOYETTE et al. further teaches dislodging microbiological content from the surfaces [0056] using an aqueous medium and adding mechanical or chemical methods or a combination thereof [0056]. An enzyme and/or a combination of enzymes can be added to the aqueous medium [0058].
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have modified the methods of modified MEYN admixing the aqueous rinsing water and enzymatic cleaner and the mixture circulating in the plant to instead an aqueous medium being 40-50 degrees before mixing the aqueous medium with the enzyme cleaner. The skilled artisan would have been motivated to select BOYETTE et al. temperature of the aqueous medium because both Modified MEYN and BOYETTE et al. achieves the same outcome., Modifying the temperature of the aqueous medium to be 40-50 degrees before mixing the aqueous medium with the enzyme cleaner (0047) as provided by BOYETTE et al., provides likewise sought functionality with reasonable expectation of success. MPEP 2143(I)(G) and allows for cell lysing [0036] as expressed by BOYETTE et al).
With respect to claim 5, Modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 1 (see above).
Modified MEYN further teaches admixing an aqueous rinsing medium and enzymatic cleaner and the mixture circulating in the plant as specified previously (MEYN, Page 4, paragraph starting with “That in the bypass cycle”). Modified MEYN further teaches the biofilms being removed enzymatically by adding an enzyme-containing cleaning agent to break up the surface of the biofilm (MEYN, Page 5, paragraph starting with “As a rule”).
However, MEYN did not teach the type of enzymes that were contained in the enzymatic cleaner. More specifically, MEYN did not teach wherein the enzymatic solution comprises one or more enzymes from the group consisting of amylases, proteases, and saccharases.
BOYETTE et al. teaches a method for quantifying microbiological content an aqueous medium as specified above [0006] and [0019]. BOYETTE et al. teaches an aqueous medium that releases intracellular content of microbiological matter into the aqueous medium by lysing the microbiological matter ([0006]). BOYETTE et al. teaches lysing the microbiological matter chemically, where chemicals are used to disrupt the cell barriers and allow intracellular content to be released ([0034]). Chemicals include detergents, enzymes, extraction solvent, and lysing buffers. BOYETTE et al. specifically teaches enzymes included but not limited to, lysozymes, mutanolysin, labiase, lysostaphin, lyticase, proteinase K, endolysin or achromopeptidases. BOYETTE et al. also teaches chemical methods for dislodging microbial cells from the surface [0056] into the aqueous medium [0056] such as adding an enzyme, examples of enzymes include, but are not limited to, blends of cellulase, alpha-amylase and protease. [0058]).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have combined the methods of Modified MEYN using an enzymatic cleaner to remove biofilm in a plant with a enzymatic solution comprising one or more enzymes from the group consisting of amylases, proteases, and saccharases because Modified MEYN uses a enzymatic cleaner to break up the surface layer of the biofilm while BOYETTE et al. achieves that same outcome. In this combination, both Modified MEYN and BOYETTE et al. are performing the same functions they would if they were separate. The person of ordinary skill in the art would have found it obvious to combine the elements because ordinarily skilled artisans would have recognized the reasons for applying Modified MEYN detection method using BOYETTE et al. enzymes and would have known how to do so. The person of ordinary skill in the art would further have predicted that the combination would remove biofilms using enzymes by lysing the cell because BOYETTE et al. teaches that the addition of enzymes can also remove the biofilm from growth surfaces. MPEP 2143 (I)(A)
With respect to claim 12, modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 2.
Modified MEYN further teaches admixing an aqueous rinsing medium and enzymatic cleaner and the mixture circulating in the plant as specified previously (see above in claim 1) (MEYN, Page 4, paragraph starting with “That in the bypass cycle”).
Modified MEYN did not specify the rinsing medium is tempered to 40-50 degrees before admixing the enzymatic solution.
BOYETTE et al. is used to remedy this, as BOYETTE et al. teaches a method for measuring the total microbiological content of an aqueous medium with the released intracellular content of the microbiological matter from an industrial plant ([0006] and [0019]). BOYETTE et al. specifically teaches removes a portion off the aqueous medium as a testing sample for biofilm detection [0024]. BOYETTE et al. specifically teaches the temperature of the aqueous medium being 40-100 degrees before removing the microbiological matter [0047]. BOYETTE et al. further teaches dislodging microbiological content from the surfaces [0056] using an aqueous medium and adding mechanical or chemical methods or a combination thereof [0056]. An enzyme and/or a combination of enzymes can be added to the aqueous medium [0058].
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have modified the methods of Modified MEYN admixing the aqueous rinsing water and enzymatic cleaner and the mixture circulating in the plant to instead an aqueous medium being 40-50 degrees before mixing the aqueous medium with the enzyme cleaner. The skilled artisan would have been motivated to select BOYETTE et al. temperature of the aqueous medium because both Modified MEYN and BOYETTE et al. achieves the same outcome., Modifying the temperature of the aqueous medium to be 40-50 degrees before mixing the aqueous medium with the enzyme cleaner [0047] as provided by BOYETTE et al., provides likewise sought functionality with reasonable expectation of success MPEP 2143(I)(G) and allows for cell lysing [0036] as expressed by BOYETTE et al.
With respect to claim 14, modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 2 (MEYN, MARTINO et al).
Modified MEYN further teaches admixing an aqueous rinsing medium and enzymatic cleaner and the mixture circulating in the plant as specified previously (see above) (MEYN, Page 4, paragraph starting with “That in the bypass cycle”). Modified MEYN further teaches the biofilms being removed enzymatically by adding an enzyme-containing cleaning agent to break up the surface of the biofilm (MEYN, Page 5, paragraph starting with “As a rule”).
However, MEYN did not teach the type of enzymes that were contained in the enzymatic cleaner. More specifically, MEYN did not teach wherein the enzymatic solution comprises one or more enzymes from the group consisting of amylases, proteases, and saccharases.
BOYETTE et al. teaches a method for quantifying microbiological content an aqueous medium as specified above [0006] and [0019]. BOYETTE teaches an aqueous medium that releases intracellular content of microbiological matter into the aqueous medium by lysing the microbiological matter ([0006]). BOYETTE et al. teaches lysing the microbiological matter chemically, where chemicals are used to disrupt the cell barriers and allow intracellular content to be released ([0034]). Chemicals include detergents, enzymes, extraction solvent, and lysing buffers. BOYETTE et al. specifically teaches enzymes included but not limited to, lysozymes, mutanolysin, labiase, lysostaphin, lyticase, proteinase K, endolysin or achromopeptidases. BOYETTE et al. also teaches chemical methods for dislodging microbial cells from the surface [0056] into the aqueous medium [0056] such as adding an enzyme, examples of enzymes include, but are not limited to, blends of cellulase, alpha-amylase and protease. [0058]).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have combined the methods of Modified MEYN using an enzymatic cleaner to remove biofilm in a plant with to a enzymatic solution comprising one or more enzymes from the group consisting of amylases, proteases, and saccharases because Modified MEYN uses a enzymatic cleaner to break up the surface layer of the biofilm while BOYETTE et al. achieves that same outcome. In this combination, both methods both Modified MEYN and BOYETTE et al. are performing the same functions they would if they were separate. The person of ordinary skill in the art would have found it obvious to combine the elements because ordinarily skilled artisans would have recognized the reasons for applying Modified MEYN detection method using BOYETTE et al. enzymes and would have known how to do so. The person of ordinary skill in the art would further have predicted that the combination would remove biofilms using enzymes by lysing the cell because BOYETTE et al. teaches that the addition of enzymes can also remove the biofilm from growth surfaces. MPEP 2143 (I)(A)
Claims 9-11 and 19, 20 are rejected under 35 U.S.C. 103 as being obvious over MEYN (DE 102017005695) in view of MARTINO et al. (Assessing chemical cleaning of nanofiltration membranes in a drinking water production plant: a combination of chemical composition analysis and fluorescence microscopy) as applied to claim 1 above and further view of REIDT et al. (Automated Immunomagnetic Processing and Separation of Legionella pneumophila With Manual Detection by Sandwich ELISA and PCR Amplification of the ompS Gene).
With respect to claim 9, modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 1.
Modified MEYN further teaches detection of biofilm using fluorescence microscopy and fluorescent markers to specifically bind to biofilm components to allow for the visualization of carbohydrate-containing extracellular polymers in biofilms (Martino et al., Page 220, “Fluorescently labelled lectins” and paragraph starting with “Samples of the fouled membranes”). More specifically, Modified MEYN teaches staining biofilm polysaccharides with lectins such as DAPI, WGA, PNA and ConA (Martino et al., Page 220, paragraph starting with “Samples of the fouled membranes”). Modified MEYN further teaches using two lectins as a double staining method (Martino et al, Page 220, paragraph starting with “Samples of the fouled membranes”).
However, Modified MEYN does not teach subjecting a part of the sample to a polymerase chain reaction process in which a specific deoxyribonucleic acid strand segment is amplified as a biofilm component.
REIDT et al. is used to remedy this, as REIDT et al. teaches a double staining method that allows for specific detection by fluorescently labeled antibody with cell staining as a viable marker (Page 158, paragraph starting with “For rapid enumeration”). REIDT et al. teaches coating paramagnetic beads coated with a specific antibody against L. pneumophila to detect bacteria (Page 159, paragraph starting with “200 mL of streptavidin-coated”). REIDT et al. also teaches that L. pneumophila is a thin-like bacteria that can lives in potable water systems (Page 158, paragraph starting with “in this article”). REIDT et al. further teaches detecting L. pneumophila by amplification of the ompS gene using PCR to make the detection process highly specific (Page 158 paragraph starting with “in this article”). REIDT et al. further teaches an internal PCR product with a length of 849 bp was amplified (Page 163, paragraph “A conventional PCR”). REIDT et al. teaches a need for fast detection of L. pneumophila with high specificity is necessity for water monitoring (Page 158, paragraph starting with “The standard detection”).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have modified the methods of Modified MEYN to stain the biofilm components with a fluorescent marker that binds to the biofilm component instead to subjecting a part of the sample to a polymerase chain reaction process in which a specific deoxyribonucleic acid strand segment is amplified as a biofilm component as taught by REIDT et al. when the outer membrane protein (ompS) was detected using PCR (Figure 5) to aid in the detection of the pathogen. Reidt et al. expresses that a number of PCR techniques for detecting L. pneumophila have been described in the literature. (Page 158, paragraph starting with “in this article”). The skilled artisan could have applied the known improvement technique in the same way to the detection method to obtain predictable results since the ompS gene is highly conserved which makes it the perfect target for the detection of the pathogen. MPEP 2143)(I)(C)
With respect to claim 10, Modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 1.
Modified MEYN teaches using fluorescence microscopy to visually observe fluorescently stained biofilm components in a food and beverage production plant (MARTINO et al., Page 220 paragraph starting with “Samples of the fouled membranes”). Modified MEYN teaches Confocal Laser Scanning Microscopy analysis of fouled membranes stained with lectins (MARTINO et al, Page 224, Figure 4). Modified MEYN teaches DAPI staining nucleic acids and polysaccharides being stained with lectins such as FITC, TRITC for polysaccharides and BSA, ConA or a mixture of PNA and WGA for double staining (MARTINO et al, Page 220 paragraph starting with “Samples of the fouled”).
Modified MEYN does not teach wherein the image of the sample is generated as part of an enzyme-linked immunosorbent detection process where the specific antibody binds to the antigen.
REIDT et al. is used to remedy this, as REIDT et al. teaches a detection method of L. pneumophila which is inhabitant of potable water systems (Page 157, paragraph starting with “Legionella pneumophila”). More specifically, REIDT et al. teaches a method where antibodies related to Legionella pneumophila are used for immunomagnetic separation and immunosorbent detection (Page 163 paragraph starting with “In this article) and (Page 163 paragraph starting with “Streptavidin beads were coated). REIDT et al. teaches paramagnetic beads coated antibodies that selectively bind to L. pneumophila cells which allow for the bacteria to be observed and isolated from the water samples (abstract).
REIDT et al. further teaches fluorescence visualization using FITC to label antibodies during the process of enzyme linked immunosorbent detection (Page 162 paragraph starting with “For direct bacterial identification” and Figure 3). .
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have modified the methods of Modified MEYN by combining the visual examination of fluorescent dyes that bind to nucleic acids and polysaccharides of bulk biofilm with the method of REIDT et al. of generating an image while using an enzyme linked detection process to specifically detect microbiological material in the biofilm. Modified MEYN teaches that generating an image of fouled membranes stained with lectins is useful for detection of biofilm components in food and beverage production plant and REIDT et al. teaches that L. pneumophila is a pathogen found in water production plants. In this combination, both Modified MEYN visual examination of lectin labelled biofilm components and REIDT et al. method of coating magnetic beads with L. pneumophila antibody and FITC for pathogen visualization and detection are performing the same functions they would if they were separate. MPEP 2143 (I)(A) The person of ordinary skill in the art would have found it obvious to combine the elements because ordinarily skilled artisans would have recognized the reasons for applying Modified MEYN visualization method using REDIT et al. antigen-antibody visualization detection method and would have known how to do so. The person of ordinarily skill in the art would further have predicted that the combination would allow for specific detection of the pathogen because REIDT et al. teaches that generating an image using antibody coated beads to detect the pathogen was effective. REIDT et al. teaches that the usage of small superparamagnetic particles or beads coated with antibodies against surface antigens in pro and eukaryotic cells has been applied for to detect a wide spread of pathogens using immunoassay (Page 158, paragraph starting with “Another technique”).
With respect to claim 11, REIDT et al. and Modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 10.
Modified MEYN teaches using fluorescence microscopy to visually observe fluorescently stained biofilm components in a production plant (MARTINO et al, Page 220 paragraph starting with “Samples of the fouled membranes”) as a detection process. Modified MEYN teaches Confocal Laser Scanning Microscopy analysis of fouled membranes stained with lectins (MARTINO et al, Page 224, Figure 4). Modified MEYN teaches DAPI staining nucleic acids and polysaccharides being stained with lectins such as FITC, TRITC for polysaccharides and BSA, ConA or a mixture of PNA and WGA for double staining (MARTINO et al, Page 220 paragraph starting with “Samples of the fouled”).
Modified MEYN does not teach wherein the enzyme-linked immunosorbent detection process is a sandwich enzyme-linked immunosorbent detection process.
REIDT et al. is used to remedy this, as REIDT et al. teaches a detection method of L. pneumophila which is inhabitant of potable water systems (Page 157, paragraph starting with “Legionella pneumophila”). More specifically, REIDT et al. teaches a method where antibodies related to Legionella pneumophila are used for immunomagnetic separation and immunosorbent detection (Page 163 paragraph starting with “In this article) and (Page 163 paragraph starting with “Streptavidin beads were coated”). REIDT et al. teaches paramagnetic beads coated antibodies that selectively bind to L. pneumophila which allow for the bacteria to be observed (abstract).
REIDT et al. further teaches fluorescence visualization using FITC to label antibodies during the process of a sandwich-based enzyme linked immunosorbent detection process (Page 162 paragraph starting with “For direct bacterial identification” and Figure 3). REIDT et al. teaches inactivated L. pneumophila captured by streptavidin-coated magnetic beads that is then binded to a FITC labeled antibody, that is conjugated to HRP (Page 159, paragraph starting with “200 µL of streptavidin-coated” and “1x107 inactivated bacteria” and Figure 1A).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have modified the methods of Modified MEYN by combining the visual examination of fluorescent dyes that bind to nucleic acids and polysaccharides of bulk biofilm for a detection method with the method of REIDT et al. of wherein the enzyme-linked immunosorbent detection process is a sandwich enzyme-linked immunosorbent detection process. Modified MEYN teaches that generating an image of fouled membranes stained with lectins is useful for detection and visual observation of biofilm components in food and beverage production plant and REIDT et al. teaches that L. pneumophila is a pathogen found in water production plants. In this combination, both Modified MEYN visual examination of lectin labelled biofilm components and REIDT et al. sandwich based ELISA detection process are performing the same functions they would if they were separate. The person of ordinary skill in the art would have found it obvious to combine the elements because ordinarily skilled artisans would have recognized the reasons for applying Modified MEYN visualization method using REDIT et al. a sandwich-based ELISA method for visualization detection method and would have known how to do so. The person of ordinarily skill in the art would further have predicted that the combination would allow for specific detection of the pathogen because REIDT et al. teaches that generating an image using antibody coated beads to detect the pathogen was effective. MPEP 2143 (I)(A) REIDT et al. teaches that the usage of small superparamagnetic particles or beads coated with antibodies against surface antigens in pro and eukaryotic cells has been applied for to detect a wide spread of pathogens using immunoassay (Page 158, paragraph starting with “Another technique”).
With respect to claim 19, REIDT et al. and Modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 9.
Modified MEYN teaches using fluorescence microscopy to visually observe fluorescently stained biofilm components in a production plant (MARTINO et al, Page 220 paragraph starting with “Samples of the fouled membranes”). Modified MEYN teaches Confocal Laser Scanning Microscopy analysis of fouled membranes stained with lectins. Modified MEYN teaches DAPI staining nucleic acids and polysaccharides being stained with lectins such as FITC, TRITC for polysaccharides and BSA, ConA or a mixture of PNA and WGA for double staining (MARTINO et al, Page 220 paragraph starting with “Samples of the fouled”). Modified MEYN teaches Confocal Laser Scanning Microscopy analysis of fouled membranes stained with lectins (MARTINO et al, Page 224, Figure 4).
Modified MEYN does not teach wherein the image of the sample is generated as part of an enzyme-linked immunosorbent detection process where the specific antibody binds to the antigen.
REIDT et al. is used to remedy this as REIDT et al. teaches a detection method of L. pneumophila which is inhabitant of potable water systems (Page 157, paragraph starting with “Legionella pneumophila”). More specifically, REIDT et al teaches a method where antibodies related to L. pneumophila are used for immunomagnetic separation and immunosorbent detection (Page 163 paragraph starting with “In this article)) and (Page 163 paragraph starting with “Streptavidin beads were coated). REIDT et al. teaches paramagnetic beads coated antibodies that selectively bind to L. pneumophila which allow for the bacteria to be observed (abstract).
REIDT et al. further teaches fluorescence visualization using FITC to label antibodies during the process of enzyme linked immunosorbent detection (Page 162 paragraph starting with “For direct bacterial identification” and Figure 3).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have combined the methods of modified MEYN by combining the visual examination of fluorescent dyes that bind to nucleic acids and polysaccharides of bulk biofilm with the method of REIDT et al. of generating an image while using an enzyme linked detection process to specifically detect microbiological material in the biofilm. Modified MEYN teaches that generating an image of fouled membranes stained with lectins is useful for detection of biofilm components in food and beverage production plant and REIDT et al. teaches that L. pneumophila is a pathogen found in water production plants. In this combination, both Modified MEYN visual examination of lectin labelled biofilm components and REIDT et al. method of coating magnetic beads with L. pneumophila antibody and FITC for pathogen visualization and detection are performing the same functions they would if they were separate. The person of ordinary skill in the art would have found it obvious to combine the elements because ordinarily skilled artisans would have recognized the reasons for applying Modified MEYN visualization method using REDIT et al. antigen-antibody visualization detection method and would have known how to do so. The person of ordinarily skill in the art would further have predicted that the combination would allow for specific detection of the pathogen because REIDT et al teaches that generating an image using antibody coated beads to detect the pathogen was effective MPEP 2143 (1)(A). REIDT et al. teaches that the usage of small superparamagnetic particles or beads coated with antibodies against surface antigens in pro and eukaryotic cells has been applied for to detect a wide spread of pathogens using immunoassay (Page 158, paragraph starting with “Another technique”).
With respect to claim 20, REIDT et al and Modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 19.
Modified MEYN teaches using fluorescence microscopy to visually observe fluorescently stained biofilm components in a production plant (MARTINO et al, Page 220 paragraph starting with “Samples of the fouled membranes”) as a detection process. Modified MEYN teaches Confocal Laser Scanning Microscopy analysis of fouled membranes stained with lectins (MARTINO et al, Page 224, Figure 4). Modified MEYN teaches DAPI staining nucleic acids and polysaccharides being stained with lectins such as FITC, TRITC for polysaccharides and BSA, ConA or a mixture of PNA and WGA for double staining (MARTINO et al, Page 220 paragraph starting with “Samples of the fouled”). Modified MEYN teaches Confocal Laser Scanning Microscopy analysis of fouled membranes stained with lectins (MARTINO et al, Page 224, Figure 4).
Modified MEYN does not teach wherein the enzyme-linked immunosorbent detection process is a sandwich enzyme-linked immunosorbent detection process.
REIDT et al. is used to remedy this, as REIDT et al. teaches a detection method of L. pneumophila which is inhabitant of potable water systems (Page 157, paragraph starting with “Legionella pneumophila”). More specifically, REIDT et al teaches a method where antibodies related to L. pneumophila are used for immunomagnetic separation and immunosorbent detection (Page 163 paragraph starting with “In this article) and (Page 163 paragraph starting with “Streptavidin beads were coated). REIDT et al. teaches paramagnetic beads coated antibodies that selectively bind to L. pneumophila cells which allow for the bacteria to be observed (abstract).
REIDT et al. further teaches fluorescence visualization using FITC to label antibodies during the process of a sandwich-based enzyme linked immunosorbent detection process (Page 162 paragraph starting with “For direct bacterial identification” and Figure 3). REIDT et al. teaches inactivated L. pneumophila captured by streptavidin-coated magnetic beads that is then binded to a FITC labeled antibody that is conjugated to HRP (Page 159, paragraph starting with “200 µL of streptavidin-coated” and “1x107 inactivated bacteria” and Figure 1A).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have combined the methods of modified MEYN by combining the visual examination of fluorescent dyes that bind to nucleic acids and polysaccharides of bulk biofilm for a detection method with the method of REIDT et al. of wherein the enzyme-linked immunosorbent detection process is a sandwich enzyme-linked immunosorbent detection process. Modified MEYN teaches that generating an image of fouled membranes stained with lectins is useful for detection and visual observation of biofilm components in food and beverage production plant and REIDT et al. teaches that L. pneumophila is a pathogen found in water production plants. In this combination, both Modified MEYN visual examination of lectin labelled biofilm components and REIDT et al sandwich-based ELISA detection process is performing the same functions they would if they were separate. The person of ordinary skill in the art would have found it obvious to combine the elements because ordinarily skilled artisans would have recognized the reasons for applying Modified MEYN visualization method using REDIT et al. a sandwich-based ELISA method for visualization detection method and would have known how to do so. The person of ordinarily skill in the art would further have predicted that the combination would allow for specific detection of the pathogen because REIDT et al. teaches that generating an image using antibody coated beads to detect the pathogen was effective (MPEP 2143) (1)(A). REIDT et al. teaches that the usage of small superparamagnetic particles or beads coated with antibodies against surface antigens in pro and eukaryotic cells has been applied for to detect a wide spread of pathogens using immunoassay (Page 158, paragraph starting with “Another technique”).
Claims 6 and 7 are rejected under 35 U.S.C. 103 as being obvious over MEYN (DE 102017005695) in view of MARTINO et al. (Assessing chemical cleaning of nanofiltration membranes in a drinking water production plant: a combination of chemical composition analysis and fluorescence microscopy) as applied to claim 1 above in further view of JINSEN et al (CN 201410037559 A).
With respect to claim 6, modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 1 (MEYN and MARTINO et al.).
Modified MEYN teaches a method for detecting biofilms in food and beverage production plants (1) for food and / or beverage production characterized by the method described (MEYN, 0001, 0007), introducing a rinsing medium into the plant when emptied after establishing a circuit of detection (MEYN, 0017). Modified MEYN teaches an aqueous rinsing medium being admixed with an enzymatic cleaner (MEYN, 0041) and circulating in plant before measuring the amount of biofilm in the mixture (MEYN, 0010).
However, modified MEYN does not teach wherein the mixture is circulated in the closed plant at constant temperature for at least one hour before the taking of the sample.
JINSEN et al. is used to remedy this as JINSEN et al. teaches a cleaning in place method that cleans milk beverage production plants (Page 2, paragraph starts with “the invention relates”) which removes dirt (a multi-layer structure comprising inorganic contaminates, protein contaminates and organic and carbohydrate substrates (Page 8, paragraph starts with “The front in”). JINSEN et al. teaches an aqueous water being introduced as the first to clean the apparatus after establishing a circuit (Page 3, paragraph starts with “Introducing water as”). JINSEN et al. teaches washing the production plant with a wash cleaner that circulates in the plant during the washing step for a duration of 20 minutes, 30 minutes, 50 minutes or 1 hour (Page 7, paragraph starts with “In one embodiment, the cleaning”). JINSEN also teaches the temperature of the cleaning solution during cleaning is from 50 to 150 ℃, preferably from 80 to 140 ℃, more preferably from 80 to 130 ℃ (Page 7, paragraph starts with “In one embodiment, the cleaning”, Embodiment 1 and 2).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have modified the methods of Modified MEYN using a cleaning mixer that circulates in a food/beverage plant before measuring the amount of biofilm in the mixture instead to a cleaning mixture that circulates the production pipes for at least an hour, tempered from 50 to 150 ℃ at the time of cleaning. Routine optimization of Modified MEYN cleaning method would have led to the claimed detection method because JINSEN et al. teaches that the constant temperature of the mixture can vary between the temperatures of from 50 to 150 ℃ at the time of cleaning and that time which can useful for the detection of dirt (Page 7, paragraph starts with “In one embodiment, the cleaning”, Embodiment 1 and 2). The person of ordinary skill in the art would have found it obvious to optimize the temperature by selecting a temperature in the range from 50 to 150 ℃ and circulating the mixture for at least an hour as suggested by JINSEN et al. JINSEN et al. teaches that the duration of each washing step is depended on the apparatus needing to clean dirt accumulation degree and the scale of the equipment (Page 7, paragraph starts with “In one embodiment, the cleaning”). MPEP 2143(I)(E) Furthermore, JINSEN et al. teaches that that is common practice to establish a cleaning process and after it is produced to determine if the cleaning process is effective and if it is not the cleaning time is changed along with the cleaning washing temperature (Page 2, paragraph starts with “After the super-high-temperature”).
With respect to claim 7, Modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 1.
Modified MEYN teaches a cleaner mixture that circulates in a closed plant (see above) (MEYN, Page 4, paragraph starting with “That in the bypass cycle”). Modified MEYN teaches a method for detecting biofilms in food and beverage production plants (1) for food and / or beverage production characterized by the method described (MEYN, 0001, 0007), introducing a rinsing medium into the plant when emptied after establishing a circuit of detection (MEYN, 0017). Modified MEYN teaches an aqueous rinsing medium being admixed with an enzymatic cleaner (MEYN, 0041) and circulating in plant before measuring the amount of biofilm in the mixture (MEYN, 0010).
However, MEYN does not teach wherein a pH value of the mixture is changed and said mixture with the changed pH value circulates in the closed plant for at least one hour before the taking of the sample.
JINSEN et al. teaches a cleaning in place method that cleans milk beverage production plants (Page 2, paragraph starts with “the invention relates”). JINSEN et al. teaches an aqueous water being introduced as the first to clean the apparatus after establishing a circuit (Page 3, paragraph starts with “Introducing water as”). JINSEN et al. teaches washing the production plant with water as the first cleaning liquid (Page 3, paragraph starts with “The invention claims”) an alkaline wash cleaner or acid wash (Page 3, paragraph starts with “The invention claims”). JINSEN et al. further teaches the alkaline cleaner in the range of 10 to 13.5 or an acidic cleaner at 0.5-3 that remove dirt (Page 7, paragraph starting with “acid cleaning solution”). JINSEN et al. also teaches quantifying the amount of dirt after the CIP process by collecting the circuited cleaning fluid. (Figs 9a and 9b). JINSEN et al. further teaches washing the production plant for the duration of 20 minutes, 30 minutes, 50 minutes or 1 hour (Page 7, paragraph starts with “In one embodiment, the cleaning”).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have combined the methods of Modified MEYN using a cleaning mixer that circulates in a food/beverage plant before measuring the amount of biofilm in the mixture with the method of JINSEN et al. wherein a pH value of the mixture is changed and said mixture with the changed pH value circulates in the closed plant for at least one hour before the taking of the sample. It would have been obvious to combine JINSEN et al. pH cleaner mixture circulating for at least an hour with Modified MEYN circulated cleaner method because Modified MEYN et al. applies to a production plant to remove biofilm while JINSEN et al. achieves that same outcome. In this combination, both Modified MEYN cleaning method and JINSEN et al. cleaning method are performing the same functions they would if they were separate. The person of ordinary skill in the art would have found it obvious to combine the elements because ordinary skilled artisans would have recognized the reasons for applying Modified MEYN cleaning method using JINSEN et al. a cleaning mixture that changes pH during circulation for at least an hour and would have known how to do so. MPEP 2143 (I)(A) The person of ordinary skill in the art would further have predicted that the combination would remove biofilm from production plants because JINSEN et al. teaches that a pH changing cleaner that circulates for at least an hour effectively removes dirt from beverage production plants.
Claims 15 and 16 are rejected under 35 U.S.C. 103 as being obvious over MEYN (DE 102017005695) in view of MARTINO et al. (Assessing chemical cleaning of nanofiltration membranes in a drinking water production plant: a combination of chemical composition analysis and fluorescence microscopy), in further view of BOYETTE et al. (US 20100112630 A1) as applied to claim 14 above and further view of JINSEN et al (CN 109158366 B).
With respect to claim 15, Modified MEYN and BOYETTE et al. teaches all of the elements of the current invention as stated above with respect to claim 14.
Modified MEYN teaches a method for detecting biofilms in food and beverage production plants (1) for food and / or beverage production characterized by the method described (MEYN, 0001, 0007), introducing a rinsing medium into the plant when emptied after establishing a circuit of detection (MEYN, 0017). Modified MEYN teaches an aqueous rinsing medium being admixed with an enzymatic cleaner (MEYN, 0041) and circulating in plant before measuring the amount of biofilm in the mixture (MEYN, 0010).
However, Modified MEYN does not teach wherein the mixture is circulated in the closed plant at constant temperature for at least one hour before the taking of the sample.
JINSEN et al. is used to remedy this, as JINSEN et al. teaches a cleaning in place method that cleans milk beverage production plants (Page 2, paragraph starts with “the invention relates”) which removes dirt (a multi-layer structure comprising inorganic contaminates, protein contaminates and organic and carbohydrate substrates (Page 8, paragraph starts with “The front in”). JINSEN et al teaches an aqueous water being introduced as the first to clean the apparatus after establishing a circuit (Page 3, paragraph starts with “Introducing water as”) JINSEN et al. teaches washing the production plant with a wash cleaner that circulates in the plant during the washing step for a duration of 20 minutes, 30 minutes, 50 minutes or 1 hour (Page 7, paragraph starts with “In one embodiment, the cleaning”). JINSEN et al. also teaches the temperature of the cleaning solution during cleaning is from 50 to 150 ℃, preferably from 80 to 140 ℃, more preferably from 80 to 130 ℃ (Page 7, paragraph starts with “In one embodiment, the cleaning”, Embodiment 1 and 2).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have modified the methods of Modified MEYN using a cleaning mixer that circulates in a food/beverage plant before measuring the amount of biofilm in the mixture instead to a cleaning mixture that circulates the production pipes for at least an hour, tempered from 50 to 150 ℃ at the time of cleaning. Routine optimization of Modified MEYN cleaning method would have led to the claimed detection method because JINSEN et al. teaches that the constant temperature of the mixture can vary between the temperatures of from 50 to 150 ℃ at the time of cleaning and that time which can useful for the detection of dirt (Page 7, paragraph starts with “In one embodiment, the cleaning”, Embodiment 1 and 2). The person of ordinary skill in the art would have found it obvious to optimize the temperature by selecting a temperature in the range from 50 to 150 ℃ and circulating the mixture for at least an hour as suggested by JINSEN et al. JINSEN et al teaches that the duration of each washing step is depended on the apparatus needing to clean dirt accumulation degree and the scale of the equipment (Page 7, paragraph starts with “In one embodiment, the cleaning”). MPEP 2143 (I)(E) Furthermore, JINSEN et al. teaches that is common practice to establish a cleaning process and after it is produced to determine if the cleaning process is effective and if it is not the cleaning time is changed along with the cleaning washing temperature (Page 2, paragraph starts with “After the super-high-temperature”).
With respect to claim 16, modified MEYN and BOYETTE et al. teaches all of the elements of the current invention as stated above with respect to claim 14.
Modified MEYN teaches a cleaner mixture that circulates in a closed plant (see above) (MEYN, Page 4, paragraph starting with “That in the bypass cycle”). Modified MEYN teaches a method for detecting biofilms in food and beverage production plants (1) for food and / or beverage production characterized by the method described (MEYN, 0001, 0007), introducing a rinsing medium into the plant when emptied after establishing a circuit of detection (MEYN, 0017). Modified MEYN teaches an aqueous rinsing medium being admixed with an enzymatic cleaner (MEYN, 0041) and circulating in plant before measuring the amount of biofilm in the mixture (MEYN, 0010).
However, MEYN does not teach wherein a pH value of the mixture is changed and said mixture with the changed pH value circulates in the closed plant for at least one hour before the taking of the sample.
JINSEN et al. teaches a cleaning in place method that cleans milk beverage production plants (Page 2, paragraph starts with “the invention relates”). JINSEN et al. teaches an aqueous water being introduced as the first to clean the apparatus after establishing a circuit (Page 3, paragraph starts with “Introducing water as”). JINSEN et al. teaches washing the production plant with water as the first cleaning liquid (Page 3, paragraph starts with “The invention claims”) an alkaline wash cleaner or acid wash (Page 3, paragraph starts with “The invention claims”) JINSEN et al. further teaches the alkaline cleaner in the range of 10 to 13.5 or an acidic cleaner at 0.5-3 that remove dirt (Page 7, paragraph starting with “acid cleaning solution”). JINSEN et al. also teaches quantifying the amount of dirt after the CIP process by collecting the circuited cleaning fluid. (Figs 9a and 9b). JINSEN et al. further teaches washing the production plant for the duration of 20 minutes, 30 minutes, 50 minutes or 1 hour (Page 7, paragraph starts with “In one embodiment, the cleaning”).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have combined the methods of Modified MEYN using a cleaning mixer that circulates in a food/beverage plant before measuring the amount of biofilm in the mixture with the method of JINSEN et al wherein a pH value of the mixture is changed and said mixture with the changed pH value circulates in the closed plant for at least one hour before the taking of the sample. It would have been obvious to combine JINSEN et al. pH cleaner mixture circulating for at least an hour with Modified MEYN circulated cleaner method because Modified MEYN et al applies to a production plant to remove biofilm while JINSEN et al. achieves that same outcome. In this combination, both Modified MEYN cleaning method and JINSEN et al. cleaning method are performing the same functions they would if they were separate. The person of ordinary skill in the art would have found it obvious to combine the elements because ordinary skilled artisans would have recognized the reasons for applying Modified MEYN cleaning method using JINSEN et al. cleaning mixture that changes pH during circulation for at least an hour and would have known how to do so. MPEP 2143 (I)(A). The person of ordinary skill in the art would further have predicted that the combination would remove biofilm from production plants because JINSEN et al teaches that a pH changing cleaner that circulates for at least an hour effectively removes dirt from beverage production plants.
Claims 4 and 13 are rejected under 35 U.S.C. 103 as being obvious over MEYN (DE 102017005695) in view of MARTINO (Assessing chemical cleaning of nanofiltration membranes in a drinking water production plant: a combination of chemical composition analysis and fluorescence microscopy) as applied to claim 1 above and further view of MOHAN et al. (JP 2021-56232 A).
With respect to claim 4, modified MEYN teaches all of the elements of the current invention as stated above with respect to claim 1.
Modified MEYN teaches using fluorescence microscopy to visually observe fluorescently stained biofilm components in a production plant (MARTINO et al, Page 220 paragraph starting with “Samples of the fouled membranes”) Modified MEYN teaches Confocal Laser Scanning Microscopy analysis of fouled membranes stained with lectins. Modified MEYN teaches DAPI staining nucleic acids and polysaccharides being stained with lectins such as FITC, TRITC for polysaccharides and BSA, ConA or a mixture of PNA and WGA for double staining (MARTINO et al, Page 220 paragraph starting with “Samples of the fouled”). Modified MEYN teaches Confocal Laser Scanning Microscopy analysis of fouled membranes stained with lectins (MARTINO et al, Page 224, Figure 4).
However, Modified MEYN does not teach wherein a proportion of organic residues in the sample is quantitatively determined before the image is generated and no detection signal is output if the proportion is below a predetermined threshold.
MOHAN et al. is used to remedy this, as MOHAN et al. teaches image processing for the identification of biological samples (Page 5, paragraph starting with “The methods”). MOHAN et al. teaches the biological sample being pretreated with binders specific to the cellular components and fluorescent dyes that can be useful for visualization (Page 19, paragraph starting with “In some embodiments, a method for microscopic”).
More specifically, MOHAN et al. teaches that quantification fluorescence microscopy of cells including intracellular and biochemical attributes such as lysosomes, mitochondria, nucleic acid and proteins (Page 19, paragraph starting with “In some embodiments, methods, systems, and instruments for quantitative microscopy are provided herein”). MOHAN et al. further teaches a cell staining method that allows for the observation by obtaining a quantitative measurement where if the measurement meets a threshold suggesting that further analysis is desired, then for further analysis, the original image that produced the quantitative result (Page 54-55, paragraph starting with “In one non-limiting example, all these”). MOHAN et al. further teaches that the information and images allow the determination of whether staining is in cells, such as in the cytoplasm, in the nucleus, in the membrane, or at the location of other organelles or other cells (Page 54-55, paragraph starting with “In one non-limiting example, all these”).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have combined the methods of modified MEYN by combining the visual examination of fluorescent dyes that bind to nucleic acids and polysaccharides of bulk biofilm for a detection method with the method of MOHAN et al. of wherein a proportion of organic residues in the sample is quantitatively determined before the image is generated and no detection signal is output if the proportion is below a predetermined threshold. Modified MEYN teaches that generating an image of fouled membranes stained with lectins is useful for detection and visual observation of biofilm components in food and beverage production plant and MOHAN et al. teaches the measurement of biological samples. In this combination, both Modified MEYN visual examination of lectin labelled biofilm components and MOHAN et al. visual examination of stained cells wherein a proportion of organic residues in the sample is quantitatively are performing the same functions they would if they were separate. MPEP 2143 (1)(A) The person of ordinary skill in the art would have found it obvious to combine the elements because ordinarily skilled artisans would have recognized the reasons for applying Modified MEYN visualization method using MOHAN et al. image threshold detection method for visualization detection method and would have known how to do so. The person of ordinarily skill in the art would further have predicted that the combination would allow for specific detection of the cellular material because MOHAN et al. teaches that generating an image wherein a proportion of organic residues in the sample is quantitatively determined before the image is generated and no detection signal is output if the proportion is below a predetermined threshold was effective.
With respect to claim 13, Modified MEYN and BOYETTE et al. teaches all of the elements of the current invention as stated above with respect to claim 2.
Modified MEYN teaches using fluorescence microscopy to visually observe fluorescently stained biofilm components in a production plant (MARTINO et al, Page 220 paragraph starting with “Samples of the fouled membranes”) Modified MEYN teaches Confocal Laser Scanning Microscopy analysis of fouled membranes stained with lectins. Modified MEYN teaches DAPI staining nucleic acids and polysaccharides being stained with lectins such as FITC, TRITC for polysaccharides and BSA, ConA or a mixture of PNA and WGA for double staining (MARTINO et al, Page 220 paragraph starting with “Samples of the fouled”). Modified MEYN teaches Confocal Laser Scanning Microscopy analysis of fouled membranes stained with lectins (MARTINO et al, Page 224, Figure 4).
However, Modified MEYN does not teach wherein a proportion of organic residues in the sample is quantitatively determined before the image is generated and no detection signal is output if the proportion is below a predetermined threshold.
MOHAN et al. is used to remedy this, as MOHAN et al. teaches image processing for the identification of biological samples (Page 5, paragraph starting with “The methods”). MOHAN et al. teaches the biological sample being pretreated with binders specific to the cellular components and fluorescent dyes that can be useful for visualization (Page 19, paragraph starting with “In some embodiments, a method for microscopic”).
More specifically, MOHAN et al. teaches that quantification fluorescence microscopy of cells including intracellular and biochemical attributes such as lysosomes, mitochondria, nucleic acid and proteins (Page 19, paragraph starting with “In some embodiments, methods, systems, and instruments for quantitative microscopy are provided herein”). MOHAN et al. further teaches a cell staining method that allows for the observation by obtaining a quantitative measurement where if the measurement meets a threshold suggesting that further analysis is desired, then for further analysis, the original image that produced the quantitative result (Page 54-55, paragraph starting with “In one non-limiting example, all these”). MOHAN et al. further teaches that the information and images allow the determination of whether staining is in cells, such as in the cytoplasm, in the nucleus, in the membrane, or at the location of other organelles or other cells (Page 54-55, paragraph starting with “In one non-limiting example, all these”).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have combined the methods of Modified MEYN by combining the visual examination of fluorescent dyes that bind to nucleic acids and polysaccharides of bulk biofilm for a detection method with the method of MOHAN et al. of wherein a proportion of organic residues in the sample is quantitatively determined before the image is generated and no detection signal is output if the proportion is below a predetermined threshold. Modified MEYN teaches that generating an image of fouled membranes stained with lectins is useful for detection and visual observation of biofilm components in food and beverage production plant and MOHAN et al. teaches the measurement of biological samples. In this combination, both Modified MEYN visual examination of lectin labelled biofilm components and MOHAN et al. visual examination of stained cells wherein a proportion of organic residues in the sample is quantitatively are performing the same functions they would if they were separate. The person of ordinary skill in the art would have found it obvious to combine the elements because ordinarily skilled artisans would have recognized the reasons for applying Modified MEYN visualization method using MOHAN et al. image threshold detection method for visualization detection method and would have known how to do so. MPEP 2143(1)(A) The person of ordinarily skill in the art would further have predicted that the combination would allow for specific detection of the cellular material because MOHAN et al. teaches that generating an image wherein a proportion of organic residues in the sample is quantitatively determined before the image is generated and no detection signal is output if the proportion is below a predetermined threshold was effective.
Claim 17 is rejected under 35 U.S.C. 103 as being obvious over MEYN (DE 102017005695) in further view of MARTINO (Assessing chemical cleaning of nanofiltration membranes in a drinking water production plant: a combination of chemical composition analysis and fluorescence microscopy) as applied to claim 4 above, in further view of MOHAN et al. (JP 2021-56232 A).
With respect to claim 17, modified MEYN and MOHEN et al. teaches all of the elements of the current invention as stated above with respect to claim 4.
MEYN teaches wherein the sample is a filter cake (surface layer of the biofilm) by an external rinsing cycle, into which the production plant or production plant part is investigated by being fluidly coupled to a measuring circuit (Page 2, paragraph starting with “ A device for carrying”). MEYN teaches particularly a heat exchanger and flow-poor areas of at least a part of the circulated mixture (Page 6, paragraph starting with “Instead of the entire” and Page 7, paragraph starting with “compared to the flow”). A rinsing cycle was introduced into the production plant/plant part coupled circuit as the first step of detecting the biofilm (Page 7, paragraph starting with “In the first process”) from the production plant/ production part.
However, MEYN does not teach specifically filter cake obtained by filtration.
MARTINO et al. teaches a cleaning in place process for nanofiltration units that are used in drinking water plants (Page 220, paragraph starting with “Nanofiltration membranes”). More specifically, MARTINO et al. teaches identifying the major components of fouling material from a filtration membrane surface (Page 220, paragraph starting with “The primary objective”). MARTINO et al. teaches foulants of several membrane sheets that were scraped for cake removal (Page 220, paragraph starting with “Foulants of several membrane”). MARTINO et al. further teaches that the nanofiltration membranes were removed from a drinking water plant that was in operation for 6 years prior detection, cleaning and analysis of the foulant components (Page 220, paragraph starting with “Nanofiltration (NF) membranes”).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to modify the removal of biofilm surface layer in a heat exchanger and/or flow dead spaces of a food and beverage plant by MEYN to instead removing a cake foulant from nanofiltration membranes that were used in a drinking water plant as taught by MARTINO et al. because both MEYN and MARTINO et al. focus on removing and detecting biofilm from plant parts specifically flow restricted areas of the production plant, because MARTINO et al. teaches removing cake from a nanofiltration unit of drinking plant membrane that were obtained after operation instead of detecting biofilm that is encapsulated by a surface layer that is removed from a food and beverage heat exchanger and/or flow dead spaces, and this involves simple substitution of the production plant to obtain predictable results. MPEP 2143 (I)(B)
Claims 18 is rejected under 35 U.S.C. 103 as being obvious over MEYN in DE 102017005695 in view of MARTINO in Assessing chemical cleaning of nanofiltration membranes in a drinking water production plant: a combination of chemical composition analysis and fluorescence microscopy, in further view of BOYETTE et al. (US 20100112630 A1) as applied to claim 14 above and further view of REIDT et al (Automated Immunomagnetic Processing and Separation of Legionella pneumophila With Manual Detection by Sandwich ELISA and PCR Amplification of the ompS Gene).
With respect to claim 18, modified MEYN and BOYETTE et al. teaches all of the elements of the current invention as stated above with respect to claim 14.
Modified MEYN further teaches detection of biofilm using fluorescence microscopy and fluorescent markers to specifically bind to biofilm components to allow for the visualization of carbohydrate-containing extracellular polymers in biofilms (Martino et al., Page 220, “Fluorescently labelled lectins” and paragraph starting with “Samples of the fouled membranes”). More specifically, Modified MEYN teaches staining biofilm polysaccharides with lectins such as DAPI, WGA, PNA and Con A (Martino et al., Page 220, paragraph starting with “Samples of the fouled membranes”). Modified MEYN further teaches using two lectins as a double staining method (Martino et al, Page 220, paragraph starting with “Samples of the fouled membranes”).
However, Modified MEYN does not teach subjecting a part of the sample to a polymerase chain reaction process in which a specific deoxyribonucleic acid strand segment is amplified as a biofilm component.
REIDT et al. is used to remedy this as REIDT et al. teaches a double staining method that allows for specific detection by fluorescently labeled antibody with cell staining as a viable marker (Page 158, paragraph starting with “For rapid enumeration”). REIDT et al. teaches coating paramagnetic beads coated with a specific antibody against L. pneumophila to detect bacteria (Page 159, paragraph starting with “200 mL of streptavidin-coated”). REIDT et al. also teaches that L. pneumophila is a thin-like bacteria that can lives can be present potable water systems (Page 158, paragraph starting with “in this article”). REIDT et al. further teaches detecting the L. pneumophila by amplification of the ompS gene using PCR to make the detection process highly specific (Page 158 paragraph starting with “in this article”). REIDT et al. further teaches an internal PCR product with a length of 849 bp was amplified (Page 163, paragraph “A conventional PCR”) REIDT et al. teaches a need for fast detection of L. pneumophila with high specificity is necessity for water monitoring (Page 158, paragraph starting with “The standard detection”).
Therefore, it would have been obvious to one of ordinary skill in the art prior to the effective filing date of the instant invention to have modify the methods of Modified MEYN to stain the biofilm components with a fluorescent marker that binds to the biofilm component instead to subjecting a part of the sample to a polymerase chain reaction process in which a specific deoxyribonucleic acid strand segment is amplified as a biofilm component as taught by REIDT et al. when the outer membrane protein (ompS) was detected using PCR (Figure 5) to aid in the detection of the pathogen. Reidt et al expresses that a number of PCR techniques for detecting L. pneumophila have been described in the literature. (Page 158, paragraph starting with “in this article”) The skilled artisan could have applied the known improvement technique in the same way to the detection method to obtain predictable results. MPEP 2143 (I)(C)
Conclusion
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/S.J.B./ Examiner, Art Unit 1758
/MARIS R KESSEL/ Supervisory Patent Examiner, Art Unit 1758