Comparison of specific and random priming in the reverse transcriptase polymerase chain reaction for genotyping group A rotaviruses.

Abstract:

:This study describes an approach to the molecular typing of rotaviruses which requires only a single RNA extraction and reverse transcription (RT) reaction using random primers. Random-primed RT provides complementary DNA (cDNA) which can be used not only for G- and P-typing polymerase chain reactions (PCR), but also for the detection of other RNA viruses which may act as enteric pathogens. It is a sensitive and specific method that can detect 10 virus particles/ml of 10% faecal suspension provided the cDNA is amplified in a nested typing-PCR. Of 121 specimens positive for rotavirus by EM and analysed using this method, only 8% could not be G- or P-genotyped. The untyped samples were tested again performing the RT reaction with G- and P-specific primers, achieving a 5% increase in sensitivity. Comparing G-genotyping against G-serotyping, 92% were genotyped through random priming RT-PCR whereas only 64% were serotyped using G-serotype specific monoclonal antibodies (MAbs).

journal_name

J Virol Methods

authors

Iturriza-Gomara M,Green J,Brown DW,Desselberger U,Gray JJ

doi

10.1016/s0166-0934(98)00168-2

subject

Has Abstract

pub_date

1999-03-01 00:00:00

pages

93-103

issue

1-2

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(98)00168-2

journal_volume

78

pub_type

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