Development of one-step real-time PCR assay for titrating trivalent live attenuated influenza vaccines

Y. Zang; D. Du; P. Ge; Y. Xu; X. Liu; Y. Zhang; W. Su; I. Kiseleva; L. Rudenko; F. Xu; W. Kong; C. Jiang

Статья

Development of one-step real-time PCR assay for titrating trivalent live attenuated influenza vaccines

Y. Zang, D. Du, P. Ge, Y. Xu, X. Liu, Y. Zhang, W. Su, I. Kiseleva, L. Rudenko, F. Xu, W. Kong, C. Jiang,

2021

https://doi.org/10.4161/hv.34453

Traditionally, infectivity of a trivalent live attenuated influenza vaccines (LAIVs) is titrated by determining the 50% egg infectious dose assay (EID50) or plaque forming units (PFU), which requires specific monoclonal antibodies to neutralize 2 strains while estimating the titer of the non-neutralized strain. Compared to this time-consuming, laborious, subjective and variable process, reverse transcription-quantitative real-time PCR (RT-qPCR) technology has advantages of rapidity, sensitivity, reproducibility and reduced contamination, thus has been applied widely for detecting pathogens andmeasuring viral titers. In this study, the critical harvest time was determined to be 18 h post-infection (hpi) for type A influenza and 12 hpi for type B influenza, but no significant difference between titers at 12 hpi and 18 hpi for the type B strain was observed. In conclusion, trivalent LAIVs can be titrated simultaneously within 24 h by this one-step RT-qPCR assay, which yielded titers comparable to those obtained by the traditional EID50 assay. Therefore, the RT-qPCR assay may be used as a highly specific, sensitive, precise and rapid alternative to the EID50 assay for titering LAIVs. © 2014 Taylor & Francis Group, LLC.

Цитирование

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Версии

1. Version of Record от 2021-04-27

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Об авторах

Y. Zang
National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, China
D. Du
Key Laboratory for Molecular Enzymology and Engineering, Ministry of Education, Jilin University, Changchun, China
P. Ge
Changchun BCHT Biotechnology Company, Changchun, China
Y. Xu
Institute of Experimental Medicine, RAMS, St. Petersburg, Russian Federation
X. Liu
Y. Zhang
W. Su
I. Kiseleva
L. Rudenko
F. Xu
W. Kong
C. Jiang

Название журнала

Human Vaccines and Immunotherapeutics

Том

Выпуск

Страницы

3642-3648

Ключевые слова

influenza vaccine; influenza vaccine; live vaccine; Article; assay; comparative study; controlled study; EID50 assay; influenza A; influenza B; measurement accuracy; real time polymerase chain reaction; reproducibility; reverse transcription polymerase chain reaction; sensitivity and specificity; virus infectivity; virus titration; human; isolation and purification; Orthomyxoviridae; procedures; real time polymerase chain reaction; standards; Humans; Influenza Vaccines; Orthomyxoviridae; Real-Time Polymerase Chain Reaction; Reproducibility of Results; Sensitivity and Specificity; Vaccines, Attenuated

Издатель

Landes Bioscience

Тип документа

journal article

Источник

scopus

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Статья

Development of one-step real-time PCR assay for titrating trivalent live attenuated influenza vaccines

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