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Development of a Taqman RT-PCR assay for the detection and quantification of negatively stranded RNA of human enteroviruses: evidence for false-priming and improvement by tagged RT-PCR.

Abstract：

:Human enteroviruses are among the most common viruses infecting humans. These viruses are known to be able to infect a wide range of tissues and are believed to establish persistent infections. Enteroviruses are positive-sense single-stranded RNA viruses whose replication involves the synthesis of negative strand intermediates. Therefore, the specific detection of negatively stranded viral RNA in tissues or cells is a reliable marker of active enteroviral replication. The present report presents the development of a real-time RT-PCR allowing the specific detection and quantification of negatively stranded viral RNA. Since it was known that specific amplification of single-stranded RNA can be made difficult by false-priming events leading to false-positive or overestimated results, the assay was developed by using a tagged RT primer. This tagged RT-PCR was shown to be able to amplify specifically negative RNA of enteroviruses grown in cell cultures by preventing the amplification of cDNAs generated by false-priming.

journal_name

J Virol Methods

journal_title

Journal of virological methods

authors

Bessaud M,Autret A,Jegouic S,Balanant J,Joffret ML,Delpeyroux F

doi

10.1016/j.jviromet.2008.07.010

subject

Has Abstract

pub_date

2008-11-01 00:00:00

pages

182-9

issue

2

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(08)00255-3

journal_volume

153

pub_type

杂志文章

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