Improvement of antigen blotting in a tissue blot immunobinding assay for the detection of two chili pepper viruses.
J Microbiol Biotechnol
; 17(11): 1885-9, 2007 Nov.
Article
em En
| MEDLINE
| ID: mdl-18092476
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.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Vírus de Plantas
/
Capsicum
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Revista:
J Microbiol Biotechnol
Ano de publicação:
2007
Tipo de documento:
Article