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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1991 Mar;29(3):519–523. doi: 10.1128/jcm.29.3.519-523.1991

Detection of group B and C rotaviruses by polymerase chain reaction.

V Gouvea 1, J R Allen 1, R I Glass 1, Z Y Fang 1, M Bremont 1, J Cohen 1, M A McCrae 1, L J Saif 1, P Sinarachatanant 1, E O Caul 1
PMCID: PMC269811  PMID: 1645368

Abstract

We adapted the polymerase chain reaction (PCR) to detect the noncultivatable group B and C rotaviruses and introduced a simple and convenient technique to purify viral RNA from stool specimens. Double-stranded RNA present in stool extracts was purified by adsorption to hydroxyapatite and was used as the template for reverse transcription and polymerase amplification. Primer pairs specific for group B (gene 8) and group C (gene 6) rotaviruses were selected to amplify group-characteristic sizes of cDNA copies readily identifiable in ethidium bromide-stained agarose gels. These primer pairs were used separately in individual PCR assays or were pooled with a primer pair specific for group A rotavirus (gene 9) in a combined PCR assay for the simultaneous detection of all three rotavirus groups. The method was very sensitive and was used to identify both human and porcine strains of group B and C rotaviruses in stool specimens. A second PCR amplification with internal group-specific primers served to increase further the sensitivity of the test and to confirm the diagnostic results obtained in the first amplification.

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Selected References

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