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Title:
METHOD FOR TESTING TARGET NUCLEIC ACID IN SAMPLE
Document Type and Number:
WIPO Patent Application WO/2022/100270
Kind Code:
A1
Abstract:
A method for testing a target nucleic acid in a sample, comprising the following steps: reacting the sample with a mixed reaction system consisting of a sgRNA-Cas system and a recombinase-mediated isothermal amplification system; and after the reaction is completed, testing testable signals generated by the reaction. The sgRNA-Cas system comprises Cas12b protein, and sgRNA targeting the target nucleic acid; and the recombination-mediated isothermal amplification system comprises primers, and single-stranded DNA reporter molecules that generate testable signals after being cleaved. When a reagent user operates, a sample adding and mixing operation of stopping the reaction in the middle and adding the reaction component of the second step is reduced, so that operation uniformity and consistency are better, facilitating the improvement of reaction precision; the operation of opening a reaction tube after the amplification reaction of a recombinant polymerase is omitted, and when the two reactions are mixed, the cis-cleavage activity of Cas protease will digest the amplification product; the amplification product thus will not be released into the environment and become a source of pollution, which is especially important for high-throughput and high-sensitivity test; and reaction sensitivity is improved.

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Inventors:
SUN YANG (CN)
WANG YALEI (CN)
ZHANG ZHUO (CN)
CHENG XUEJIA (CN)
Application Number:
PCT/CN2021/118280
Publication Date:
May 19, 2022
Filing Date:
September 14, 2021
Export Citation:
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Assignee:
BEIJING SYNSORTECH CO LTD (CN)
ZHEJIANG SYNSORBIO TECH CO LTD (CN)
BEIJING SYNSORBIO TECH CO LTD (CN)
International Classes:
C12Q1/6844
Domestic Patent References:
WO2020006036A12020-01-02
Foreign References:
CN111630163A2020-09-04
CN110551800A2019-12-10
CN111593145A2020-08-28
CN109837328A2019-06-04
CN109055499A2018-12-21
CN110184329A2019-08-30
Other References:
TENG, FEI ET AL.: "CDetection: CRISPR-Cas12b-based DNA detection with sub-attomolar sensitivity and single-base specificity", GENOME BIOLOGY, vol. 20, 1 July 2019 (2019-07-01), XP055817045, ISSN: 1474-760X, DOI: 10.1186/s13059-019-1742-z
Attorney, Agent or Firm:
BEIJING TANGSONG IP FIRM (CN)
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