Visual loop-mediated isothermal amplification (LAMP) method for the rapid test of tobacco

• Main Authors: ZHANG, Ke, ZHANG, Xiaowei, CHEN, Dan, TONG, Zhijun, WANG, Chunqiong, QIN, Shichun, YANG, Jinchu, SUN, Haowei, LI, Haiyan, LONG, Jie
• Format: Patent
• Language: English
• Published: 30.05.2024
• 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; SPIRITS; VINEGAR; WINE

A visual loop-mediated isothermal amplification (LAMP) method for a rapid test of tobacco includes: extracting a genomic DNA of the tobacco, designing primers, establishing a LAMP reaction system, and optimizing the LAMP reaction system. The present disclosure designs the LAMP primers according to a conserved region of a screened tobacco-specific Ntsp151 genomic sequence, and establishes the LAMP method based on color determination. The present disclosure can rapidly identify flue-cured tobacco leaves, finished cigarettes, and tea cigarettes, and achieves a detection limit of 200 copies per reaction. The test results of the LAMP method for non-tobacco samples are negative, indicating that the LAMP method has high specificity. Compared with the quantitative polymerase chain reaction (qPCR) method, the LAMP method has a coincidence rate of 96.7%, but the LAMP method only needs about 1 h to obtain the results.