DETAILED ACTION
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 .
Claims Status
Claims 1, 2, 4-12, & 15-21 filed on 01/27/2026 are pending. All the amendments and arguments have been thoroughly reviewed but are deemed insufficient to place this application in condition for allowance. The following rejections are either newly applied, as necessitated by amendment, or are reiterated. They constitute the complete set being presently applied to the instant application. Response to Applicant’s argument follow. This action is FINAL.
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office Action.
Any rejection not reiterated is hereby withdrawn in view of the amendments to the claims.
Claim Rejections - 35 USC § 103
Claim(s) 1, 2, 16, & 21 is/are rejected under 35 U.S.C. 103 as being unpatentable over Amer (Amer & Almajhdi; Molecular and Cellular Probes, Vol. 25, pages 101-107, March 2011), in view of Ahrberg (Ahrberg, Manz, Neuzil; Scientific Reports, Vol. 5, pages 1-7, June 2015), as cited in the IDS dated 05/16/2022.
Regarding amended claim 1, Amer teaches a method for SYBR Green I based (single-signal fluorescent material) real-time RT-PCR assay that uses sets of primers designed specifically to amplify sequences of the nucleocapsid gene of BCoV (target nucleic acid) and an internal control sequence through repeated cycles of DNA denaturation, annealing, and extension of the BCoV (target nucleic acid) and the internal control sequence with varying specific temperatures for each cycle (continuous temperature control) (abstract lines 1-6; pg. 102-103 paragraph bridging pg. 102 & pg. 103 lines 1-17; Table 2). Amer also teaches that the fluorescence signal from SYBR Green (single-signal fluorescent material) is measured at the end of each DNA denaturation, annealing, and extension step (fluorescent signal measured at regular intervals from the start of heating for DNA denaturation to the completion of DNA denaturation) and that identification of the internal control signal was achieved through a melting curve analysis where the distinct melting temperature Tm (Tx) of 82.54 ± 0.32 was obtained (fluorescence signal measure at time point Tx is a fluorescent signal measures at a time point at which a temperature higher than the melting point of an amplified product of the internal control sequence is reached) (abstract lines 4-6; pg. 102-103 paragraph bridging pg. 102 & pg. 103 lines 1-24; pg. 105 column 1 2nd full paragraph lines 1-11). In addition, Amer teaches that the internal control primers were designed to validate the reaction steps for the qPCR amplification and detection and to confirm that the reaction was performed correctly and to exclude false-negatives (pg. 105 column 1 2nd full paragraph lines 1-6). Finally, Amer teaches the fluorescence signal from SYBR Green (single-signal fluorescent material) is measured at the end of each DNA denaturation, annealing, and extension step in which the fluorescent signal measured at the start of heating comprises a fluorescent signal of the amplified product of BCoV (target nucleic acid) and the internal sequence according to the melt curve analysis and that the fluorescent signal measured at Tx (Tm), Tm of the amplified product of the internal control sequence of 82.54 ± 0.32 was obtained, comprises only the fluorescent signal of the amplified internal control sequence (pg. 102-103 paragraph bridging pg. 102 & pg. 103 lines 1-17; pg. 105 column 1 2nd full paragraph lines 1-11; pg. 105 paragraph bridging column 1 & 2 lines 1-7).
Amer does not teach that the fluorescent signal measured at the time point Tx (Tm) comprises only the fluorescent signal of the amplified product of the target sequence.
Ahrberg teaches a method for quantitative PCR using intercalating dyes (single-signal fluorescent material) of two sequences, HA and NA, and internal controls and further conducting a melting curve analysis between the two sequences in which the HA sequence amplicon (target sequence of interest) has a melting temperature of 76°C and the second NA sequence amplicon (control sequence) has a melting temperature of 68°C (pg. 4 4th full paragraph lines 1-8). Ahrberg also teaches that differentiation of amplicons (encompassing target and internal control sequences) can be used to differentiate them (fluorescent signal measured at Tm (Tx) of above 68°C comprises only the fluorescent signal of the HA sequence amplicon (amplified product of the target sequence) (excluding the fluorescent signal of the amplified product of the NA sequence amplicon (control sequence) from the measured fluorescent signals, such that only the fluorescent signal of the amplified product of the target sequence is detected or quantified)) and that different melting temperatures can be achieved by primer design and would allow for different amplicons to be detected in the same reaction (pg. 5 1st full paragraph lines 1-7; pg. 5 2nd full paragraph lines 1-5). Further, Ahrberg teaches through the HA sequence amplicon (target sequence of interest) having a melting temperature of 76°C results in its own amplification curve enabling differentiation of the amplicons (encompassing target and internal control sequences) (excluding fluorescence signal of the amplified product of the internal control sequence such that only the fluorescence signal of the amplified product of the target sequence is detected or quantified) (pg. 4 4th full paragraph lines 1-6; Figure 3).
Amer and Ahrberg are considered to be analogous to the claimed invention because they are all in the same field of quantitative nucleic acid amplification with an intercalating dye. Therefore, it would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of detecting a fluorescent signal comprising a fluorescent signal of the amplified product of the target and the internal sequence and detecting a fluorescent signal that comprises only the fluorescent signal of the amplified internal control sequence in Amer to incorporate the detection of a fluorescent signal that comprises only the amplified product of the target sequence of interest (fluorescence signal measured at the time point Tx is of the amplified product of the target sequence) as taught in Ahrberg because Ahrberg teaches that different melting temperatures can be achieved through primer design and that this allows for the detection of multiple different amplicons in the same reaction.
Regarding claim 2, Amer teaches that the single-signal fluorescent material is the intercalating dye SYBR Green (abstract lines 1-6; pg. 102-103 paragraph bridging pg. 102 & pg. 103 lines 1-17).
Regarding claim 16, Amer teaches that the internal control sequence was designed to confirm that the reaction was performed correctly and to exclude false-negatives through detection of the internal control through a distinct melting temperature Tm (checking whether the internal control sequence is amplified or not) (abstract lines 4-6; pg. 102-103 paragraph bridging pg. 102 & pg. 103 lines 18-24; pg. 105 column 1 2nd full paragraph lines 1-11).
Regarding new claim 21, Amer teaches a method for SYBR Green I based (single-signal fluorescent material) real-time RT-PCR assay that uses sets of primers designed specifically to amplify sequences of the nucleocapsid gene of BCoV (target nucleic acid) and an internal control sequence through repeated cycles of DNA denaturation, annealing, and extension of the BCoV (target nucleic acid) and the internal control sequence with varying specific temperatures for each cycle (continuous temperature control) (abstract lines 1-6; pg. 102-103 paragraph bridging pg. 102 & pg. 103 lines 1-17; Table 2). Amer also teaches that the fluorescence signal from SYBR Green (single-signal fluorescent material) is measured at the end of each DNA denaturation, annealing, and extension step (fluorescent signal measured at regular intervals from the start of heating for DNA denaturation to the completion of DNA denaturation) and that identification of the internal control signal was achieved through a melting curve analysis where the distinct melting temperature Tm (Tx) of 82.54 ± 0.32 was obtained (fluorescence signal measure at time point Tx is a fluorescent signal measures at a time point at which a temperature higher than the melting point of an amplified product of the internal control sequence is reached) (abstract lines 4-6; pg. 102-103 paragraph bridging pg. 102 & pg. 103 lines 1-24; pg. 105 column 1 2nd full paragraph lines 1-11). In addition, Amer teaches that the internal control primers were designed to validate the reaction steps for the qPCR amplification and detection and to confirm that the reaction was performed correctly and to exclude false-negatives (pg. 105 column 1 2nd full paragraph lines 1-6). Finally, Amer teaches the fluorescence signal from SYBR Green (single-signal fluorescent material) is measured at the end of each DNA denaturation, annealing, and extension step in which the fluorescent signal measured at the start of heating comprises a fluorescent signal of the amplified product of BCoV (target nucleic acid) and the internal sequence according to the melt curve analysis and that the fluorescent signal measured at Tx (Tm), Tm of the amplified product of the internal control sequence of 82.54 ± 0.32 was obtained, comprises only the fluorescent signal of the amplified internal control sequence (pg. 102-103 paragraph bridging pg. 102 & pg. 103 lines 1-17; pg. 105 column 1 2nd full paragraph lines 1-11; pg. 105 paragraph bridging column 1 & 2 lines 1-7).
Amer does not teach that the fluorescent signal measured at the time point Tx (Tm) comprises only the fluorescent signal of the amplified product of the target sequence.
Ahrberg teaches a method for quantitative PCR using intercalating dyes (single-signal fluorescent material) of two sequences, HA and NA, and internal controls and further conducting a melting curve analysis between the two sequences in which the HA sequence amplicon (target sequence of interest) has a melting temperature of 76°C and the second NA sequence amplicon (control sequence) has a melting temperature of 68°C (pg. 4 4th full paragraph lines 1-8). Ahrberg also teaches that differentiation of amplicons (encompassing target and internal control sequences) can be used to differentiate them (fluorescent signal measured at Tm (Tx) of above 68°C comprises only the fluorescent signal of the HA sequence amplicon (amplified product of the target sequence) enabling the fluorescence signal measured at time point Tx is used to confirm whether a false negative of the PCR reaction is generated) and that different melting temperatures can be achieved by primer design and would allow for different amplicons to be detected in the same reaction (pg. 5 1st full paragraph lines 1-7; pg. 5 2nd full paragraph lines 1-5).
Amer and Ahrberg are considered to be analogous to the claimed invention because they are all in the same field of quantitative nucleic acid amplification with an intercalating dye. Therefore, it would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of detecting a fluorescent signal comprising a fluorescent signal of the amplified product of the target and the internal sequence and detecting a fluorescent signal that comprises only the fluorescent signal of the amplified internal control sequence at a melting temperature Tm (Tx) to confirm that the reaction was performed correctly and to exclude false-negatives in Amer to incorporate the detection of a fluorescent signal that comprises only the amplified product of the target sequence of interest (fluorescence signal measured at the time point Tx is of the amplified product of the target sequence) in order to confirm if a false negative is generated as taught in Ahrberg because Ahrberg teaches that different melting temperatures can be achieved through primer design and that this allows for the detection of multiple different amplicons in the same reaction.
Claim(s) 4-12, & 17-20 is/are rejected under 35 U.S.C. 103 as being unpatentable over Amer (Amer & Almajhdi; Molecular and Cellular Probes, Vol. 25, pages 101-107, March 2011) and Ahrberg (Ahrberg, Manz, Neuzil; Scientific Reports, Vol. 5, pages 1-7, June 2015), as cited in the IDS dated 05/16/2022, as applied to claims 1, 2, & 16 above, and further in view of Yeon (KR 20180090955A), machine translation obtained from Science & Technical Information Center (STIC), as cited on the IDS dated 05/16/2022.
The teachings of Amer and Ahrberg with respect to claim 1 are discussed above.
Regarding claim 4, Amer teaches that DNA denaturation cycles requires heating (first temperature at which DNA denaturation is performed) the PCR solution (reaction vessel) and then annealing cycles requires cooling (second temperature at which annealing is performed) the PCR solution (reaction vessel), and finally the extension cycles requires heating the PCR solution (reaction vessel) some but not back to the original temperature (pg. 102-103 paragraph bridging pg. 102 & pg. 103 lines 1-17).
Amer and Ahrberg does not teach wherein the continuous temperature control in step (a) comprises heating a reaction vessel to a first temperature with a heating block and then cooling to a second temperature by removing the reaction vessel from the heating block and then exposing the separated reaction vessel to an artificial air flow for a predetermined period of time.
Yeon teaches a temperature control method for nucleic acid amplification reaction in which a denaturation step is performed at a first temperature and a hybridization (annealing)/extension step is performed at a second temperature comprising a first step of heating a reaction vessel containing a PCR solution with a heating block to the first temperature and then cooling the reaction vessel to the second temperature by separating the vessel and the heating block and exposing the reaction vessel to an artificial air flow for a predetermined period of time (claim 1; claim 6; paragraph [0015] lines 1-4; paragraph [0019] lines 1-5; paragraph [0035] lines 1-5). Yeon also teaches that this method reduces the overall time required for the PCR amplification which plays an important role in enabling rapid response to rapidly spread infectious diseases and virus outbreaks and minimizing the time consumption in the temperature variation period of the nucleic acid amplification technique (paragraph [0010] lines 1-7).
Amer, Ahrberg, and Yeon are considered to be analogous to the claimed invention because they are all in the same field of nucleic acid amplification. Therefore, it would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of heating and cooling the PCR solution in the DNA denaturation, annealing, and extension steps of the real-time RT-PCR assay in Amer to incorporate heating with a heating block for the DNA denaturation step and then removing the PCR solution (reaction vessel) from the heating block and exposing to an artificial air flow to cool to a temperature (second temperature) for DNA annealing as taught in Yeon because Yeon teaches that doing so would provide a method to reduce the time consumption in the temperature variation period of the nucleic acid amplification technique and enable a more rapid response to detecting diseases and viruses.
Regarding claim 5, Amer teaches that DNA denaturation cycles requires heating (first temperature at which DNA denaturation is performed) the PCR solution (reaction vessel) and then annealing cycles requires cooling (third temperature at which annealing is performed) the PCR solution (reaction vessel), and finally the extension cycles requires heating (second temperature at which the DNA extension is performed) the PCR solution (reaction vessel) some but not back to the original temperature (pg. 102-103 paragraph bridging pg. 102 & pg. 103 lines 1-17).
Amer and Ahrberg does not teach wherein the continuous temperature control in step (a) comprises heating a reaction vessel to a first temperature with a heating block and then cooling to a third temperature by removing the reaction vessel from the heating block and then exposing the separated reaction vessel to an artificial air flow for a predetermined period of time and then heating the cooled reaction vessel to a second temperature by bringing the cooled reaction vessel in contact with the heating block and then separating the reaction vessel from the heating block.
Yeon teaches a temperature control method for nucleic acid amplification reaction in which a denaturation step is performed at a first temperature, a hybridization (annealing) step is performed at a second hybridization temperature, and an extension step is performed at a second extension temperature comprising a first step of heating a reaction vessel containing a PCR solution with a heating block to the first temperature (temperature where DNA denaturation is performed), then cooling the reaction vessel to the second hybridization temperature (temperature where hybridization (annealing) is performed) by separating the vessel and the heating block and exposing the reaction vessel to an artificial air flow for a predetermined period of time, and then heating the container to a second extension temperature (temperature where extension is performed) by contacting the reaction vessel with the heating block and then removing the reaction vessel from the heating block (claim 8; paragraph [0058] lines 1-7). Yeon also teaches that this method reduces the overall time required for the PCR amplification which plays an important role in enabling rapid response to rapidly spread infectious diseases and virus outbreaks and minimizing the time consumption in the temperature variation period of the nucleic acid amplification technique (paragraph [0010] lines 1-7).
Amer, Ahrberg, and Yeon are considered to be analogous to the claimed invention because they are all in the same field of nucleic acid amplification. Therefore, it would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of heating and cooling the PCR solution in the DNA denaturation, annealing, and extension steps of the real-time RT-PCR assay in Amer to incorporate heating with a heating block for the DNA denaturation step and then removing the PCR solution (reaction vessel) from the heating block and exposing to an artificial air flow to cool to a temperature (second hybridization (annealing) temperature) for DNA annealing and then contacting the PCR solution (reaction vessel) with the heating block and then separating again to a temperature (second extension temperature) for DNA extension as taught in Yeon because Yeon teaches that doing so would provide a method to reduce the time consumption in the temperature variation period of the nucleic acid amplification technique and enable a more rapid response to detecting diseases and viruses.
Regarding claim 6, Yeon teaches the position of the heating block is fixed and the container (PCR solution in a reaction vessel) moves upwards to a predetermined spacing to the heating block (reaction vessel and heating block are separated) (claim 1; claim 2).
Regarding claim 7, Yeon teaches that the reaction vessel is cooled through artificial airflow that is continuously provided (claim 1; claim 3).
Regarding claim 8, Yeon teaches the position of the container (PCR solution in a reaction vessel) is fixed and the heating block is moved downward to a predetermined position to have a predetermined interval (reaction vessel and heating block are separated) (claim 1; claim 4).
Regarding claim 9, Yeon teaches the artificial air flow is provided only when the heating block is spaced apart from the container (PCR solution in a reaction vessel) by a predetermined position (artificial air flow is only supplied when the heating block is separated from the reaction vessel) (claim 1; claim 4; claim 5).
Regarding amended claim 10, Yeon teaches that the predetermined time is determined by the general Formula 1 of t = 4 + 2*e-(v-7.4)/6.2 (claim 1; claim 7).
Regarding claim 11, Yeon teaches that the predetermined interval is 0.5 to 2 cm (the reaction vessel and the heating block are spaced by a distance of 0.5 to 2 cm) (claim 1; claim 14).
Regarding claim 12, Yeon teaches a denaturation step is performed at a first temperature, a hybridization (annealing) step is performed at a second hybridization temperature (second temperature), and an extension step is performed at a second extension temperature (fourth temperature) comprising a first step of heating a reaction vessel containing a PCR solution with a heating block to the first temperature (temperature where DNA denaturation is performed), then cooling the reaction vessel to the second hybridization temperature (second temperature where hybridization (annealing) is performed) by separating the vessel and the heating block and exposing the reaction vessel to an artificial air flow for a predetermined period of time, and then heating the container to a second extension temperature (fourth temperature where extension is performed) by contacting the reaction vessel with the heating block and then removing the reaction vessel from the heating block (claim 8; paragraph [0058] lines 1-7).
Regarding claim 17, Yeon teaches the position of the heating block is fixed and the container (PCR solution in a reaction vessel) moves upwards to a predetermined interval to the heating block (reaction vessel and heating block are separated) (claim 8; claim 9).
Regarding claim 18, Yeon teaches the position of the vessel (PCR solution in a reaction vessel) is fixed and the heating block is moved downward to a predetermined position to have a predetermined interval (reaction vessel and heating block are separated) (claim 8; claim 11).
Regarding amended claim 19, Yeon teaches that the predetermined time is determined by the general Formula 1 of t = 4 + 2*e-(v-7.4)/6.2 (claim 8; claim 13).
Regarding claim 20, Yeon teaches that the predetermined interval is 0.5 to 2 cm (the reaction vessel and the heating block are spaced by a distance of 0.5 to 2 cm) (claim 8; claim 14).
Claim(s) 15 is/are rejected under 35 U.S.C. 103 as being unpatentable over Amer (Amer & Almajhdi; Molecular and Cellular Probes, Vol. 25, pages 101-107, March 2011) and Ahrberg (Ahrberg, Manz, Neuzil; Scientific Reports, Vol. 5, pages 1-7, June 2015), as cited in the IDS dated 05/16/2022, as applied to claims 1, 2, & 16 above, and further in view of Mondal (Mondal & Venkataraman; Journal of Biochemical and Biophysical Methods. Vol. 65, pages 97-105, October 2005).
The teachings of Amer and Ahrberg with respect to claim 1 are discussed above.
Regarding claim 15, Amer and Ahrberg does not teach that the interval in step (b) is 0.5 to 1 second.
Mondal teaches a method for real time PCR detection based on fluorescence from intercalating dyes such as SYBR Green I (single-signal fluorescent material) and that the SYBR Green I fluorescent signal was measured every 0.5 seconds within a cycle of amplification (abstract lines 1-4; pg. 99-100 paragraph bridging pg. 99 & pg. 100 lines 1-2). Mondal also teaches that this method of measuring the fluorescent signal at an interval of 0.5 seconds can individually optimize the extension times of each PCR cycle while the reaction is in progress thus reducing the total number of cycles and total time required to reach maximum fluorescence compared to conventional PCR (abstract lines 5-10).
Amer, Ahrberg, and Mondal are considered to be analogous to the claimed invention because they are all in the same field of quantitative nucleic acid amplification with an intercalating dye. Therefore, it would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of measuring the fluorescent signal at an interval of after each cycle of amplification in Amer to incorporate measuring the fluorescent signal at an interval of 0.5 seconds as taught in Mondal because Mondal teaches that doing so would reduce the number of cycles and total time required to reach maximum fluorescence compared to conventional PCR.
Response to Arguments
The response traverses the rejection. The response asserts that Ahrberg fails to teach or suggest detecting or quantifying the fluorescence signal of the amplified product of the target sequence by excluding the fluorescence signal of the amplified product of the internal control sequence from the combined signals including the fluorescence signal of the amplified product of the internal control sequence and the fluorescence signal of the amplified product of the target sequence. Further, the response asserts that even though Ahrberg teaches or suggests the differentiation between the HA sequence (target sequence) amplicon and the NA amplicon (internal control sequence) based on melting curve analysis, Ahrberg does not teach or suggest that the differentiation between the HA sequence and the NA amplicon leads to the claimed feature of excluding the fluorescence signal of the amplified product of the internal control sequence from the combined fluorescence signals such that only the fluorescence signal of the amplified product of the target sequence is detected or quantified, and therefore, amended claim 1 is patentably distinguishable from the cited prior art references. These arguments have been thoroughly reviewed but were not found persuasive as Ahrberg teaches that differentiation of amplicons (encompassing target and internal control sequences) can be used to differentiate them (fluorescent signal measured at Tm (Tx) of above 68°C comprises only the fluorescent signal of the HA sequence amplicon (amplified product of the target sequence) (excluding the fluorescent signal of the amplified product of the NA sequence amplicon (control sequence) from the measured fluorescent signals, such that only the fluorescent signal of the amplified product of the target sequence is detected or quantified)) and that different melting temperatures can be achieved by primer design and would allow for different amplicons to be detected in the same reaction (pg. 5 1st full paragraph lines 1-7; pg. 5 2nd full paragraph lines 1-5). Further, Ahrberg teaches through the HA sequence amplicon (target sequence of interest) having a melting temperature of 76°C results in its own amplification curve enabling differentiation of the amplicons (encompassing target and internal control sequences) (excluding fluorescence signal of the amplified product of the internal control sequence such that only the fluorescence signal of the amplified product of the target sequence is detected or quantified) (pg. 4 4th full paragraph lines 1-6; Figure 3). Therefore, Ahrberg does teach that the differentiation between the HA sequence and the NA amplicon and the the claimed feature of excluding the fluorescence signal of the amplified product of the internal control sequence from the combined fluorescence signals such that only the fluorescence signal of the amplified product of the target sequence is detected or quantified as discussed further above.
The response also asserts that applicant has introduced independent claim 21 which, in part, recites wherein the fluorescence signal at the time point Tx is used to confirm whether a false negative of the PCR reaction is generated, and to put it another way, the fluorescence signal of the amplified product of the target sequence is used to confirm whether a false negative of the PCR reaction is generated. Further, the response asserts that among the cited prior art references, Ahrberg is silent as to the false negative of the PCR reaction and Yeon fails to teach or suggest it. Further, the response asserts that Amer teaches or suggests that the internal control sequence can be used to confirm whether a false negative of the PCR reaction is generated, however, nevertheless the prior art references are silent as to the claimed feature that the fluorescence signal measured at time point Tx, which is the amplified product of the target sequence, is used to confirm whether a false negative of the PCR reaction is generated and, therefore, claim 21 is patentably distinguishable from the cited prior art references. These arguments have been thoroughly reviewed but were not found persuasive. First, Amer teaches that identification of the internal control signal was achieved through a melting curve analysis where the distinct melting temperature Tm (Tx) of 82.54 ± 0.32 was obtained (abstract lines 4-6; pg. 102-103 paragraph bridging pg. 102 & pg. 103 lines 1-24; pg. 105 column 1 2nd full paragraph lines 1-11). In addition, Amer teaches that the internal control primers were designed to validate the reaction steps for the qPCR amplification and detection and to confirm that the reaction was performed correctly and to exclude false-negatives (pg. 105 column 1 2nd full paragraph lines 1-6). Further, Ahrberg teaches a method for quantitative PCR using intercalating dyes (single-signal fluorescent material) of two sequences, HA and NA, and internal controls and further conducting a melting curve analysis between the two sequences in which the HA sequence amplicon (target sequence of interest) has a melting temperature of 76°C and the second NA sequence amplicon (control sequence) has a melting temperature of 68°C (pg. 4 4th full paragraph lines 1-8). Ahrberg also teaches that differentiation of amplicons (encompassing target and internal control sequences) can be used to differentiate them (fluorescent signal measured at Tm (Tx) of above 68°C comprises only the fluorescent signal of the HA sequence amplicon (amplified product of the target sequence) enabling the fluorescence signal measured at time point Tx is used to confirm whether a false negative of the PCR reaction is generated) and that different melting temperatures can be achieved by primer design and would allow for different amplicons to be detected in the same reaction (pg. 5 1st full paragraph lines 1-7; pg. 5 2nd full paragraph lines 1-5). Further, Amer and Ahrberg are considered to be analogous to the claimed invention because they are all in the same field of quantitative nucleic acid amplification with an intercalating dye. Therefore, it would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of detecting a fluorescent signal comprising a fluorescent signal of the amplified product of the target and the internal sequence and detecting a fluorescent signal that comprises only the fluorescent signal of the amplified internal control sequence at a melting temperature Tm (Tx) to confirm that the reaction was performed correctly and to exclude false-negatives in Amer to incorporate the detection of a fluorescent signal that comprises only the amplified product of the target sequence of interest (fluorescence signal measured at the time point Tx is of the amplified product of the target sequence) in order to confirm if a false negative is generated as taught in Ahrberg because Ahrberg teaches that different melting temperatures can be achieved through primer design and that this allows for the detection of multiple different amplicons in the same reaction. Therefore, the combination of prior art references of Amer and Ahrberg teach the claimed feature that the fluorescence signal measured at time point Tx, which is the amplified product of the target sequence, is used to confirm whether a false negative of the PCR reaction is generated.
The response also asserts that claims 2, 4-12, & 15-20 depend from claim 1, and thus, they are also patentable over the cited prior art references. These arguments have been thoroughly reviewed but were not found persuasive for the reasons set forth above.
For these reasons, and the reasons already made of record and modified to address the claims as currently amended, the rejections are maintained and applied to the newly amended claims.
Conclusion
Claims 1, 2, 4-12, & 15-21 are rejected.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to BAILEY C BUCHANAN whose telephone number is (703)756-1315. The examiner can normally be reached Monday-Friday 8:00am-5:00pm ET.
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/BAILEY BUCHANAN/Examiner, Art Unit 1682
/JEHANNE S SITTON/Primary Examiner, Art Unit 1682
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