METHOD FOR DETECTING THE CSH 1 COLD SHOCK GENE IN VIBRIO CHOLERAE STRAINS USING REAL-TIME PCR Russian patent published in 2022 - IPC C12N15/00 C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention relates to the field of biotechnology. A method for detecting the csh1 cold shock gene in Vibrio cholerae strains using PCR is presented, including DNA isolation from the strain under study, performing PCR with specific primers CSh 1 and CSh, followed by taking into account the amplification reaction. At the same time, 0.5 µl of the intercalating dye SYBR Green I is added to the incubation mixture to identify amplification products with subsequent melting of the formed amplicons, and according to their melting temperature, in the range of 72-94°C and according to the dynamics of changes in the fluorescence index through the FAM channel of the amplifier, the results are recorded according to the geometric method by registering the C_p index. In the presence of the target product, the cshl cold shock gene, an amplificon with low C_p is formed, and in the absence of a specific target, the reaction with a pair of CSh 1- CSh 2 primers forms other amplificons with high C_p.

EFFECT: invention makes it possible to use it in laboratory diagnostics when monitoring cholera and conducting studies related to the study of the characteristics of the biology of the pathogen, including the identification of genetic structures that ensure the adaptation of cholera to adverse environmental factors.

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