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Detection of Rotaviruses by Nucleic Acid Hybridization with Cloned DNA of Simian Rotavirus SA11 Genes

D. H. Dimitrov, D. Y. Graham and M. K. Estes
The Journal of Infectious Diseases
Vol. 152, No. 2 (Aug., 1985), pp. 293-300
Published by: Oxford University Press
Stable URL: http://www.jstor.org/stable/30104679
Page Count: 8
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Detection of Rotaviruses by Nucleic Acid Hybridization with Cloned DNA of Simian Rotavirus SA11 Genes
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Abstract

We developed a dot-blot hybridization assay to detect rotaviral RNA sequences in tissue culture or in clinical samples. $^{32}P-labeled$ cloned cDNA probes of the simian rotavirus SA11 specifically detected rotaviral RNA sequences and were more sensitive for detecting SA11 than was the commercial enzyme-linked immunosorbent assay $Rotazyme^®$ test. A full-length probe of SA11 gene 6 detected $2.5 \times 10^{5}$ SA11 particles or $\sim 0.27 ng$ of purified SA11 dsRNA. Combined probes from genes 6 and 9 detected 0.135 ng of purified SA11 dsRNA. The assay detected group A rotaviruses from different subgroups and serotypes, but the sensitivity of RNA detection varied from 0.5 to 31 ng when RNA from heterologous strains of virus was analyzed. An analysis of coded stool samples correctly identified 31 (91%) of 34 samples positive for rotavirus by electron microscopy and 100% of 26 samples negative for rotavirus by electron microscopy. Preliminary experiments also showed the assay has potential to directly characterize (subgroup and serotype) rotaviral isolates.

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