Abstract

  Yam mosaic virus (YMV) is one of the most economically important virus infecting yams. Immunoassays are routinely used for laboratory diagnosis of YMV and for certification of planting materials. However, YMV antibodies, the key reagents, needed for these immunoassays are not readily available. We describe in this paper, the production of YMV monoclonal antibodies for the detection of YMV. The monoclonal antibody was produced by immunizing six weeks old BALB/c mice with YMV hybridoma cells and tapping soft peritoneal tumor tissues for antibody. Antibody titre was determined by triple antibody sandwich-enzyme-linked immunosorbent assay (TAS-ELISA) using YMV infected yam leaves and non-infected tissue culture yam leaves. The antibody produced had a titre of 1:1,310,720 and an optimal TAS-ELISA detection dilution of 1:80,000. This high-titre YMV monoclonal antibody is useful for monitoring and certification purposes.   Key words: Monoclonal antibodies, ascetic fluid, yam mosaic virus.

Highlights

  • Millions of people in Africa depend on yam for food each day (FAO, 2009)

  • We describe in this paper, the production of Yam mosaic virus (YMV) monoclonal antibodies for the detection of YMV

  • This paper reports the production of a sensitive, high titre YMV monoclonal antibody in mice peritoneum

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Summary

Production of yam mosaic virus monoclonal antibodies in mice peritoneum

Yam mosaic virus (YMV) is one of the most economically important virus infecting yams. Immunoassays are routinely used for laboratory diagnosis of YMV and for certification of planting materials. YMV antibodies, the key reagents, needed for these immunoassays are not readily available. We describe in this paper, the production of YMV monoclonal antibodies for the detection of YMV. The monoclonal antibody was produced by immunizing six weeks old BALB/c mice with YMV hybridoma cells and tapping soft peritoneal tumor tissues for antibody. Antibody titre was determined by triple antibody sandwich-enzyme-linked immunosorbent assay (TAS-ELISA) using YMV infected yam leaves and non-infected tissue culture yam leaves. The antibody produced had a titre of 1:1,310,720 and an optimal TAS-ELISA detection dilution of 1:80,000. This high-titre YMV monoclonal antibody is useful for monitoring and certification purposes

INTRODUCTION
Production of ascetic fluid
Antibody titration
Titration of YMV ascetic fluid
Healthy Diseased Detection factor
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