POLYMERASE CHAIN REACTION FOR CHARACTERIZATION OF VP4 GENE TYPES OF GROUP A BOVINE ROTAVIRUS DIRECTLY IN THE FECES OF NEONATAL DAIRY CALVES | ||||
| Veterinary Medical Journal (Giza) | ||||
| Volume 46, Issue 4, October 1998, Page 471-478 PDF (2.62 MB) | ||||
| DOI: 10.21608/vmjg.1998.377331 | ||||
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| Authors | ||||
| HUSSEIN A. HUSSEIN* 1; A. SILIM2; M.A SHALABY1; Y. EL-AZHARY2 | ||||
| 1Department of Virology, faculty of veterinary medicine, Cairo university, Giza 1221 , Egypt | ||||
| 2Department of Virology, faculty of veterinary medicine, university of montreal, st-Hyacinthe ,Quebec J25 7C6, Canada | ||||
| Abstract | ||||
| A PCR (polymerase chain reaction) genotyping assay was applied to identify bovine rotavirus (BRV) gene 4 types (P genotypes, NCDV (PI), UK (P5), and B223 (P11) in the field samples collected from calves with and without diarrhoea. Rotavirus double Stranded RNAs (dsRNAs) extracted from the fecal samples were reverse transcribed and amplitied by PCR using two nucleotide primers that correspond highly conserved regions of the VP4+ gene among rotaviruses. The amplified products (that included the VP8 portion and the connecting peptide of the VP4 gene) were then amplified by PCR int he Presence of coctail of specific primers for three reference BRV. The PCR-typing assay was applied to identify BRV P genotypes (PI, P5 and P11) in 73 field samples. These samples were reacted positive for BRV by polyacrylamide gel electrophoresis and monoclonal antibody - based ELISA, The PCR assay was able to identify BRV Different P genotypes in 79.4% of the samples. Mix of two P genotypes was efficiently detected in The same samples by applied method .in order to detect all G/P possible combination types. among BRV field isolates, the P typing results were compared with the previously reported G-typing assay applied in this study provided a very sensitive method for genotyping of Brv field strains. | ||||
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