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Improvement of Antigen Blotting in a Tissue Blot Immunobinding Assay for the Detection of Two Chili Pepper Viruses  

Han, Jung-Heon (Center for Plant Molecular Genetics and Breeding Research, Seoul National University)
Shin, Jun-Sung (Center for Plant Molecular Genetics and Breeding Research, Seoul National University)
Kim, Young-Ho (Department of Agricultural Biotechnology and Center for Plant Molecular Genetics and Breeding Research, Seoul National University)
Kim, Byung-Dong (Department of Plant Science and Center for Plant Molecular Genetics and Breeding Research)
Publication Information
Journal of Microbiology and Biotechnology / v.17, no.11, 2007 , pp. 1885-1889 More about this Journal
Abstract
The tissue blot immunobinding assay (TBIA) is widely used for the detection and localization of plant viruses in various plant tissues. The basic experimental procedures of TBIA sampling and blotting were simplified using commercially available micropipette tips. This method was termed the ring-blot immunobinding assay (R-BIA), as the blot on the membrane forms a ring shape. The detection efficacy of R-BIA was tested for two chili pepper viruses, pepper mild mottle tobamovirus (PMMoV) and pepper mottle potyvirus (PepMoV), following the optimized serological procedures of TBIA (length of the incubation period and BSA concentration, and primary and secondary antibodies). Sensitivity of the R-BIA was about 1 ng/ml of purified PMMoV in pepper leaf sap from a healthy pepper plant. R-BIA also showed high specificity in the detection of PMMoV and PepMoV. Moreover, the modified sampling and blotting procedures were simpler and more reliable than other TBIA methods (such as whole-leaf blotting and crushed-leaf blotting), suggesting that the R-BIA may be used for medium- to large-scale detection of plant viruses in laboratories with minimal facilities.
Keywords
Chili pepper; detection; pepper viruses; ring-blot; tissue blot immunobinding assay;
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Times Cited By Web Of Science : 1  (Related Records In Web of Science)
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