UNSPECIFIED (1991) DETECTION OF GROUP-B AND GROUP-C ROTAVIRUSES BY POLYMERASE CHAIN-REACTION. JOURNAL OF CLINICAL MICROBIOLOGY, 29 (3). pp. 519-523. ISSN 0095-1137

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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 hydrodroxyapatite 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.

Item Type:	Journal Article
Subjects:	Q Science > QR Microbiology
Journal or Publication Title:	JOURNAL OF CLINICAL MICROBIOLOGY
Publisher:	AMER SOC MICROBIOLOGY
ISSN:	0095-1137
Date:	March 1991
Volume:	29
Number:	3
Number of Pages:	5
Page Range:	pp. 519-523
Publication Status:	Published
URI:	http://wrap.warwick.ac.uk/id/eprint/22857

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