Method for indirect determination of concentration of glycoprotein of arriah strain of fixed rabies virus in raw materials for vaccines

A PCR-based method using specific primers and probes effectively determines glycoprotein concentration in rabies virus raw materials, addressing sensitivity and specificity issues, achieving rapid and accurate analysis.

RU2864787C1Active Publication Date: 2026-06-29FEDERALNOE GOSUDARSTVENNOE BIUDZHETNOE UCHREZHDENIE FEDERALNYI TSENTR OKHRANY ZDOROVIA ZHIVOTNYKH FGBU VNIIZZH
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Patent Information

Application Number
RU2025125321
Authority / Receiving Office
RU · RU
Patent Type
Patents
Current Assignee / Owner
Filing Date
2025-09-15
Publication Date
2026-06-29
Estimated Expiration
2045-09-15

AI Technical Summary

Technical Problem

Existing methods for determining the concentration of glycoprotein in rabies virus raw materials for vaccines, such as ELISA, are limited by sensitivity, specificity, and contamination risks, and do not effectively separate native proteins from biologically inactive fragments.

Method used

A PCR-based method using oligonucleotide primers and a molecular probe to indirectly determine the concentration of glycoprotein in rabies virus raw materials, involving RNA extraction, reverse transcription, and real-time PCR with a developed linear function to calculate glycoprotein concentration based on the threshold amplification cycle.

Benefits of technology

The method reduces analysis time to 3 hours, increases sensitivity to 0.01 μg/ml, ensures high specificity, and provides a reliable correlation between glycoprotein concentration and threshold amplification cycle, with an amplification efficiency of 99.48% and approximation reliability of 0.997.

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Abstract

FIELD: biotechnology.SUBSTANCE: quality control of raw materials in the manufacture of a culture-based inactivated anti-rabies vaccine, namely a method for indirectly determining the concentration of the glycoprotein of the ARRIAH strain of the fixed rabies virus in raw materials for vaccines. The main advantages of the proposed invention are the ability to reduce the analysis time of virus-containing suspensions when determining the concentration of the glycoprotein of the ARRIAH strain of the rabies virus in the raw material for the vaccine to 3 hours; by using original highly specific primers and a molecular probe, to achieve analytical sensitivity of up to 0.01 μg / ml; to increase the reliability of the analysis by establishing a relationship between the concentration of the glycoprotein of the ARRIAH strain of the fixed rabies virus (CRVG) and the threshold amplification cycle (Ct), presented as a linear function CRVG= –0.228Ct+ 7.615, where CRVG is the concentration of the glycoprotein of the ARRIAH strain of the fixed rabies virus, Ct is the threshold amplification cycle with high approximation reliability (R2 = 0.997) and amplification efficiency (E) of 99.48%.EFFECT: proposed model allows for indirect determination of the concentration of the glycoprotein of the ARRIAH strain of the rabies virus in vaccine raw materials using the developed method.2 cl, 1 dwg, 6 tbl, 6 ex
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Citation Information

Patent Citations

  • Method for spectrometric determination of concentration of rabies virus ribonucleoprotein by estimating the number of viral RNA molecules in raw materials for rabies vaccines

    RU2748475C1

  • Method for the indirect determination of the concentration of rabies virus ribonucleoprotein in the raw material for the vaccine during the study of the maximum extremums of the graphs of the second derivative for the logistic curves of the amplification reaction in real time

    RU2775745C1