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 .
This action is in response to applications correspondence mailed 11/3/2025. The amendment to the claims mailed 11/30/2025 has been entered.
Election/Restrictions
Applicant’s election without traverse of group I, claims 1-6 in the reply filed on 11/3/2025 is acknowledged.
Claims 7-8 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 11/3/2025.
Drawings
The drawings are acceptable.
Claim Objections
Claim 1 is objected to because of the following informalities: claim 1 recites “preparation of a magnetic probe:”, “electrode modification:”, “identification of the magnetic beads:”, “strand displacement amplification (SDA) isothermal amplification:”, “cutting:”, and “photoelectrochemical detection”. Each of the phrasings prior to the colon in steps 1-6 is not necessary and not grammatically correct for the claims. Applicant should remove each of the phrases above. Appropriate correction is required.
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.
Claims 1-6 are 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.
The claims are generally narrative and indefinite, failing to conform with current U.S. practice. They appear to be a literal translation into English from a foreign document and are replete with grammatical and idiomatic errors. It is unclear if claim 1 is directed to a method of detecting T2 toxin, method of generating a standard curve or if the claims are directed to a method of making a photoelectrochemical sensing detection system. There are no active steps that detect or identify T2 toxin in a sample. The steps of claim 1 encompass preparing a magnetic probe, preparing an electrode, separating cDNA, obtaining different concentrations of cDNA and mixing with electrode, and generating a standard curve. It is unclear what the claim is intended to encompass.
The process steps of claim 1 are indefinite. It is unclear how each of the steps are related and what each of the steps accomplish. For example, step 1 requires mixing magnetic beads with T2-APT, step 3 requires adding T2-APT for incubation and combination then adding complementary T2-cDNA to compete with a target and obtaining competitive cDNA through magnetic separation. It is unclear if T2-APT is bound to magnetic beads from step 1 and how this will compete with a target to obtain competitive cDNA. Step 2 requires preparing a CdS-Au-UCNP/ITO electrode by reducing sulfhydryl modified DNA-UCNPs and TCEP, however TCEP is used with a sulfhydryl modified DNA to attach DNA to UCNP. It is unclear if the claim is intended for DNA to be attached to UCNP or remove DNA from UCNPs to obtain a CdS-Au-UCNP/ITO electrode. Step 4 and 5 require performing strand displacement amplification with different concentrations of cDNA and incubating with CAS14a-sgRNA complex however this will generate a standard curve and does not detect an unknown amount of target. Step 3 does not requires a known amount of competitive target or cDNA. It is unclear how competing with a target and adding complementary T2-cDNA to obtain competitive cDNA is used in the assay. The step of plotting a standard curve is not related to the other process steps and it is unclear how this step is related to the method. The body of the claim does not fully or intrinsically set forth all the limitations recited within the preamble. Accordingly the preamble of the claim and the recited active process steps renders the claim indefinite because the claim does not reasonably apprise one of ordinary skill in the art the scope of the claims and the metes and bounds of the claim are unclear and it would not be readily apparent if one was infringing on the claimed invention.
Claim 1 recites the limitation of “T2-APT” and “T2-cDNA”. Claim 4 recites “TemDNA”. It is unclear what is encompassed by T2-APT, T2-cDNA, and TemDNA. None of these terms are art recognized terms to identify magnetic probe or DNA. Neither the claims or the specification provide a definition for these terms. The specification does not provide a definition for the terms. The specification states T2-aptamer (T2-APT) (para 8), complementary DNA of T2 (T2-cDNA) (para 9), biotin-modified aptamer (T2-APT) and complementary T2-cDNA is added to compete with target (See para 46) but none of these recitations provide an explicit definition. Additionally the specification provide two different embodiments for T2-APT, one comprising biotin modified aptamer and the other T2-aptamer. There is no definition or description of TemDNA. There is a table in the specification, table 1, that lists sequences for T2-APT, T2-cDNA, and TemDNA however these are not definitions and it is unclear if the sequences recited in table 1 are to be limited to the recitation of T2-APT, T2-cDNA, and TemDNA. As such it is unclear what is encompassed by the recitation of T2-APT, T2-cDNA and TemDNA and one of ordinary skill in the art would not be apprised of the metes and bounds of the instant claims.
Claim 2 is indefinite because it is unclear what is required for the preparation of the magnetic probe. The claim requires 100 µl of magnetic beads, separating and washing, adding 500µl of T2-APT and mixing at room temperature. Claim 2 depends from claim 1 and it is unclear what “the magnetic beads” are referencing. Claim 1 requires mixing magnetic beads with T2-APT to prepare the magnetic beads. It is unclear if the beads are the magnetic beads before mixing with T2-APT or are the magnetic beads after mixing with T2-APT.
Claim 4 contains the trademark/trade name CutSmart. Where a trademark or trade name is used in a claim as a limitation to identify or describe a particular material or product, the claim does not comply with the requirements of 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph. See Ex parte Simpson, 218 USPQ 1020 (Bd. App. 1982). The claim scope is uncertain since the trademark or trade name cannot be used properly to identify any particular material or product. A trademark or trade name is used to identify a source of goods, and not the goods themselves. Thus, a trademark or trade name does not identify or describe the goods associated with the trademark or trade name. In the present case, the trademark/trade name is used to identify/describe a buffer and, accordingly, the identification/description is indefinite.
Claim 5 recites the limitation "the incubating in the step (5)" in line 2 of the claim. There is insufficient antecedent basis for this limitation in the claim. Claim 5 depends from claim 1 and there is no incubating step in claim 1 or in step (5) of claim 1.
Claim 6 recites the limitation "the electrolyte in step (6)" in line 2 of the claim. There is insufficient antecedent basis for this limitation in the claim. Claim 6 depends from claim 1 and claim 1 does not require an electrolyte in step 6 or any other steps of the claim.
Conclusion
The following is relevant art cited by the examiner:
Wang (CN106680346A) teaches a method of biotin labeled aflatoxin B1 aptamer and a block probe attached to a magnetic bead. Wang teaches in the presence of a target, AFB1 and block probe are combined with the aptamer in a competitive manner, the replaced block probe triggers strand displacement amplification reaction to produce amplicons and the amplicons participate in a surface proximity hybridization reaction on the surface of an electrode to produce an electric signal (see abstract). Wang does not teach CdS-Au/ITO electrode, Cas14 and sgRNA or T2 toxin.
Xia et al. (CN105606574 A) teaches a method for detecting T2 toxin. Xia teaches the aptamer T2 toxin sequence, SEQ ID NO 1 which is identical to the aptamer T2 sequence in table 1 of the instant specification. Xia does not teach a CdS-Au/ITO electrode, use of magnetic beads, strand displacement amplification, or Cas14a and sgRNA.
Lin (WO2019010930A1) teaches upconversion material based photoelectrochemical DNA sensor and detection method. Lin teaches a photoelectrochemical DNA sensor based on upconversion material for detecting a concentration of a target ssDNA. Lin teaches photoelectrochemical DNA sensor with a reference electro of Ag/AgCL, counter electrode is platinum and substrate of the working electrode is ITO. Lin teaches target ssDNA is dropped on the working electrode fixed with the probe ssDNA for hybridization reaction. Lin does not teach the use of magnetic probe, standard curve, or Cas14a-sgRNA complex.
Zhang (Anal. Chem. 2018, 90, 11892-11898) teaches DNA strand displacement reaction on magnetic beads and detection by photoelectrochemical detection of RNA in samples. Zhang teaches magnetic beads were mixed with separation DNA at different concentrations and was collected and kept for further use (see Au NP-coupled ETSD reaction on MB). Zhang does not teach the use of CRISPR or CdS-Au-UCNP/ITO electrode.
Mao (Talanta 2022, 242, 123232, pp. 1-7) teaches the use of upconversion nanoparticle probes, UCNP with CRISPR-Cas12a enzyme to detect ochratoxin A. Mao teaches ochratoxin A aptamer was constructed and competes with OTA toxin to combine and release the activation chain which is recognized by CRISPR.Cas12A complex, which degrades ssDNA. Mao teaches a competitive response, magnetic separation and detection (see scheme 1). Mao does not teach a photoelectrochemical sensor to detect the ssDNA from the CRISPR/Cas12A complex. Mao does not teach Cas14a. Mao does not teach CdS-Au-UCNP/ITO.
Hu (Chem Comm, 2021, 57, 10423) teaches a Cas14 sensing platform due to ssDNA recognition ability and high substrate designability. Hu teaches in the presence of a target, aptamer will recognize and bind AMP molecule, releases the activator which will hybridize with crRNA and activate collateral cleavage activity of Cas14. Hu teaches the detection system comprises a biotinylated ssDNA, after cleavage is captured by magnetic beads and removed from supernatant, which is added to ICP-MS (see scheme 1). Hu does not teach analysis of the ssDNA by photoelectrochemical sensing or strand displacement amplification.
Luo (Anal Chem, 2018, 90, 9568-9575) teaches upconversion nanospheres comprising Au@CdS@UNCP for photoelectrochemical enzyme immunoassay. Luo teaches synthesis of UCNP@Au@CdS upconversion nanospheres (see experimental section). Luo teaches immunoreaction followed by photoelectrochemical detection (see pg. 9570, 1st column). Luo does not teach analysis of nucleic acids.
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/SARAE L BAUSCH/Primary Examiner, Art Unit 1699