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DETECTION OF GROUP-B AND GROUP-C ROTAVIRUSES BY POLYMERASE CHAIN-REACTION
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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 | ||||
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Subjects: | Q Science > QR Microbiology | ||||
Journal or Publication Title: | JOURNAL OF CLINICAL MICROBIOLOGY | ||||
Publisher: | AMER SOC MICROBIOLOGY | ||||
ISSN: | 0095-1137 | ||||
Official Date: | March 1991 | ||||
Dates: |
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Volume: | 29 | ||||
Number: | 3 | ||||
Number of Pages: | 5 | ||||
Page Range: | pp. 519-523 | ||||
Publication Status: | Published |
Data sourced from Thomson Reuters' Web of Knowledge
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