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

We developed a dot-blot hybridization assay to detect rotaviral RNA sequences in tissue culture or in clinical samples. 32P-labeled cloned cDNA probes of the simian rotavirus SAll specifically detected rotaviral RNA sequences and were more sensitive for detecting SAll ttian was the commercial enzyme-linked immunosorbent assay Rotazyme(R) test. A full-length probe of SAll gene 6 detected 2.5 x 10' SAll particles or r',0.27 ng of purified SAll dsRNA. Combined probes from genes 6 and 9 detected 0.135 ng of purified SAll 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 100010 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.