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Detection of Group and Subgroup Specific Antigens of Bovine Rota Viruses in Bangladesh

MUZAHED UDDIN AHMED
Department of Medicine, Bangladesh Agricultural University, Mymensingh

MD. MAHBUB ALAM
Department of Medicine, Bangladesh Agricultural University, Mymensingh

Abstract/ Executive Summary:

Faecal specimens from 150 calves with diarrhea in Savar dairy farm, Dhaka in 1992 were analysed for rotaviruses. Thirty three specimens were positive for group A rotavirus antigen by ensyme linked immunosorbent assay. In subgrouping out of 33 group A positive specimens 21 reacted with subgroup I-specific monoclonal antibody. Twelve specimens did not react with either of the subgroup I and subgroup II-specific monoclonal antibodies.

Keywords: Rotavirus, ELISA, Group A, Subgroup I and Subgroup II

Location: Department of Medicine, Bangladesh Agricultural University, Mymensingh

Start Date: 00-00-1992

End Date: 00-00-1992

Program Area: Animal Health and Management

Commodity: Calf

Objectives:

To characterize the group and subgroup specific antigens of bovine rotaviruses by ELISA using group and subgroup specific monoclonal antibodies from dairy calves in Bangladesh.

Methodology:

Faecal specimens were collected from calves during monthly visits to Savar dairy farm. Faecal specimens were kept in sterile tube, labeled and kept in -20°C. Each specimen was prepared as a 10% suspension in PBS (pH 7.4) and centrifuged at 3000 rpm for 30 minutes. The supernatants were separated and used for ELISA. The group specific Mab Yo-156, subgroup I-specific Mab S2-37 and subgroup II-specific Mab Yo-5 were used in the ELISA which was performed as follows. Briefly, polyvinyl microtitre plates were coated with 1: 1 0,000 dilution of mouse ascitic fluid containinig Mab and incubated overnight at 4°C. After washing the wells three times with PBS (pH 7.4) wells were incubated with 1 % bovine serum albumin over niqht at 4°C and subsequently washed. A mixture of 10% stool suspension (375 µl) and 10% skim milk (125 µl) was allowed to react in the wells overnight at 4°C. After being washed, 50 µl of rabbit anti human rofavirus serum (1: 10,000 dilution) was added and incubated for 1 hour at 37°C. After washing, 50 µl of peroxidase conjugated goat antirabbit immunoglobin G and added. The plates were incubated for 1 hour at 37°C and washed. The reaction with the substrate OPO was allowed to develop for 30 minutes at room temperature and then stabilized by addition of 25 µl of 3 N sulphuric acid. The reaction was evaluated by visual observation by comparing internsity of the colour visible in the wells of group and subgroups positive and group and subgroup negative wells.

Source of Information: BAU Res. Prog. 7: 488-491; 1993

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The presence of at least two antigenic determinants on VP6 has been confirmed with monoclonal antibodies. One determinant is common to all rotavirus strains and the second determinant reacts with viruses having one of the two well characterized subgroup specificities. All bovine strains that react in this assay are subgroup I viruses, while strains from human have been found to be subgroup I and II.

Knowledge Management: Report/Proceedings