Reagent for detecting replicable lentivirus by using three-channel double-quenching probe and detection method thereof

• Main Authors: CHEN JIANFENG, SUI LILI, SHEN QINWEN, CHEN HUI, WANG HANYU, KONG LINGJIE
• Format: Patent
• Language: Chinese; English
• Published: 22.10.2021
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROCESSES USING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a reagent for detecting replicable lentivirus by a three-channel double-quenching probe, which comprises a first primer probe pair, a second primer probe pair and a third primer probe pair, the first primer probe pair is a VSV-G primer and a probe pair; the second primer probe pair is a gag primer probe pair; and the third primer probe pair is an RPP30 primer probe pair. The invention further discloses a detection method of the replicable lentivirus. The detection method comprises the following steps: S1, culturing cells; S2, preparing a sample to be detected; S3, preparing a standard curve sample; S4, determining the required number of reaction holes; S5, preparing a required mixed solution of a qPCR reaction solution and a primer probe; S6, adding the mixed solution, the standard substance, the diluent, the negative quality control NCS, the to-be-detected sample and the sample ERC into the reaction holes according to requirements; S7, controlling the reaction process of each reaction