METHOD FOR IDENTIFYING GENETIC MATERIAL OF FLAVIVIRUSES TRANSMITTED BY TICKS AND MOSQUITOES BY REAL-TIME RT-PCR USING FLUORESCENT-LABELLED PROBES Russian patent published in 2025 - IPC C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention refers to biotechnology, molecular biology and virology. What is presented is a method for identifying a genetic material of flaviviruses transmitted by mosquitoes and ticks by real-time RT-PCR with using a fluorescent-labelled probe. What is used are constructed pan-specific primers for the NS5 protein gene of flaviviruses of the family Flaviviridae and fluorescence-labelled probes specific for representatives of the genus Flavivirus and flaviviruses transmitted by ticks and mosquitoes, respectively: primers: primer F-FVFL: 5´- ACAACATGATGGGRAARAGRGARAA-3´; primer R1-FV: 5´- CCARTGGTCYTCATTYAGGAATCC-3´; probe specific for classical flaviviruses: PROBE_gen-FV: FAM- GCCATCTGGTWCATGTGGCTKGG-BHQ1; probe specific for tick-borne flaviviruses: PROBE_tick-FV: HEX-TGGGGAGCCGSTTYCTGGAGTTTGA-BHQ1; probe specific for mosquito-borne flaviviruses: .PROBE_mosq-FV: FAM-AGTTTGGAAARGCGAARGGAAGCAGWGCC-BHQ1, where FAM is carboxyfluorescein, a fluorescent dye whose absorption wavelength is 492 nm and the fluorescence wavelength is 520 nm; where HEX is hexachlorofluorescein, a fluorescent dye whose absorption wavelength is 533 nm and the fluorescence wavelength is 549 nm; BHQ1 is a fluorescence quencher with a quenching range of 480-580 nm.

EFFECT: development of primers and probes for identification of genetic material of flaviviruses associated with ticks and mosquitoes, by RT-PCR with hybridisation-fluorescent detection in real time.

1 cl, 4 dwg, 4 ex