METHOD FOR INDIRECT DETERMINATION OF TITRE OF INFECTIOUS ACTIVITY OF VACCINE STRAIN "PB-97" OF RABIES VIRUS IN RAW MATERIAL FOR VACCINE BASED ON ANALYSIS OF MELTING CURVES OF HIGH-RESOLUTION AMPLICONS USING THIRD GENERATION INTERCALATING DYE EVAGREEN Russian patent published in 2024 - IPC C12N7/00 C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention relates to production of an attenuated cultural rabies vaccine. Disclosed is a method for indirect determination of titre of infectious activity of vaccine strain "PB-97" of rabies virus in raw material for vaccine based on analysis of melting curves of high-resolution amplicons using intercalating dye of the third generation EvaGreen.

EFFECT: main advantages of the proposed invention are reduce time of analysis of virus-containing suspensions for determination of titre of infectious activity of strain "PB-97" of rabies virus in raw material for vaccine to 2,5 hours; reduce the cost of one study by 29%; increase analytical sensitivity to 0,2 lg CCID₅₀/cm³ or 1,58 CCID₅₀/cm³ due to development of original highly specific primers and application of intercalating dye of the third generation EvaGreen; increase the reliability of the analysis due to the establishment of the relationship between the titre of the infectious activity of the vaccine strain "PB-97" of the rabies virus in the raw material for the vaccine and the quantification cycle—EvaGreen, presented in the form of a logarithmic function with high accuracy of approximation 0,9977 and amplification efficiency of 99,77%.

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