Molecular diagnosis and genotyping of Bovine viral diarrhea virus

Document Type : Original Article

Authors

1 Department of Virology- Faculty of veterinary medicine – University of Sadat City- Egypt.

2 Department of animal medicine and infectious diseases - Faculty of veterinary medicine - University of Sadat City- Egypt.

Abstract

In the present study isolation, biotyping, antigenic and genomic characterization of BVDV isolates was carried out on buffy coat samples obtained from apparently healthy cattle. The samples were inoculated in MDBK cells for three successive passages and the CPE was recorded indicating that 48%, 62% and 52% were positive CPE in the 1st, 2nd and 3rd passages respectively. The viral antigen was detected in cell culture after propagation also by direct FAT and reveled that (62%) showed positive result. The mean virus titer was recorded after each passage showing values of <102,102 and 104 TCID50/ml by 1st, 2nd and 3rd passages respectively. The virus was identified by VNT using the reference anti-BVDV-1 serum on cell cultures infected fluids at the third passage. It was found that all samples showed a characteristic CPE of BVDV were neutralized by the used antiserum confirming that they are BVDV-1. Nine buffy coat samples were selected to carry out the nested RT-PCR for detection and genotyping of suspected BVDV. The results indicated the presence of BVDV genotype-I. The positive samples for FAT, VNT and RT- PCR in infected cell culture were stained by H&E for biotyping indicating the presence of the two biotypes (CP& NCP) in the samples. The SNT was carried out on serum samples from the same animals indicating the prevalence of antibodies in 62% by mean titer ranged from < ½ to 1/8. In conclusion, the present study reports the identification of BVDV genotype –I in apparently healthy cattle and presence of P.I. animals.

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