Method for detecting HPV16/HPV18 type nucleic acid based on nicking enzyme coupling rolling circle amplification technology

• Main Authors: FANG YANI, LIU JINHUI
• Format: Patent
• Language: Chinese; English
• Published: 15.09.2023
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention relates to a method for detecting nucleic acid, in particular to a method for detecting HPV16/HPV18 type nucleic acid based on a nicking enzyme coupling rolling circle amplification technology. The technical problems that a traditional detection method is inaccurate in result, high in experimental condition and operation technical requirements and high in cost are solved. The method comprises the following steps: 1) designing and synthesizing an amplification primer and a single-stranded circular DNA template; 2) extracting DNA of a to-be-detected sample; 3) preparing a reaction system; the reaction system comprises a to-be-detected sample DNA and a reaction reagent combination; the reaction reagent combination is an HPV16 type reaction reagent combination and/or an HPV18 type reaction reagent combination; (4) carrying out rolling circle replication isothermal amplification reaction on the reaction system to obtain a product; and 5) detecting a product of the isothermal amplification reaction by