METHOD FOR TICK-BORNE ENCEPHALITIS VIRUS DETECTION BY REAL-TIME RT-PCR METHOD Russian patent published in 2021 - IPC C12Q1/68 C12Q1/6876 

FIELD: physics.

SUBSTANCE: invention relates to a method for detecting tick-borne encephalitis virus by RT-PCR in real time. Method comprises extracting RNA from biological samples followed by a two-step reverse transcription reaction and polymerase chain reaction for target fragments of RNA genome of tick-borne encephalitis virus using a set of oligonucleotide primers and corresponding fluorescent labeled probe, complementary to gene of NS3 gene of tick-borne encephalitis. Analysis of results is carried out using software of the device used to perform PCR with hybridisation-fluorescence detection in real-time mode, results are interpreted on the basis of presence or absence of intersection of the fluorescence curve with a threshold line installed at the corresponding level, which corresponds to the presence or absence of the value of the threshold cycle Ct. Result is considered to be positive if the fluorescence accumulation curve for the corresponding sample has a characteristic "sigmoid" shape and crosses the threshold line. Following set of oligonucleotide primers and a fluorescent labelled probe is used for detection: TBE-fsh CTg gACA TgC ACC CAg gCT, rt-TBEV-f12b AgT gTT ggA CAT gCA CCC Ag, TBE-rsh ATGCATTGGCGRATGAGCTC, rt-TBEV-r12c CTC CAT TTC CTT gAg CAC CAC, TBEv-Prb12 R6G-Tgg gAA gAC CCA CAg AgT CCT CCC-BHQ1.

EFFECT: invention provides higher diagnostic sensitivity.

1 cl, 1 dwg, 4 tbl