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Detection and quantitation of group A rotaviruses by competitive and real-time reverse transcription-polymerase chain reaction.

Abstract：

:A competitive reverse transcription-polymerase chain reaction (RT-PCR) was developed to detect and to quantitate the RNA of group A rotaviruses. In the assay, a 433 bp fragment is amplified by a one-tube RT-PCR protocol using primers with binding sites located in a highly conserved region of segment 6 of the rotavirus genome. An in vitro synthesized RNA with a 43-base deletion with respect to the wild-type sequence of this fragment was used as an internal control. Using these transcripts as templates, 10 RNA molecules were amplified reproducibly and detected in ethidium bromide-stained agarose gels or by fluorimetry using the SYBR Green I dye in a real-time RT-PCR assay. The efficiency of the protocol was confirmed by the detection of small amounts of viral RNA of group A rotaviruses in clinical samples obtained from various animal species and man.

journal_name

J Virol Methods

journal_title

Journal of virological methods

authors

Schwarz BA,Bange R,Vahlenkamp TW,Johne R,Müller H

doi

10.1016/s0166-0934(02)00118-0

subject

Has Abstract

pub_date

2002-09-01 00:00:00

pages

277-85

issue

2

eissn

0166-0934

issn

1879-0984

pii

S0166093402001180

journal_volume

105

pub_type

杂志文章

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