Studies on a Nigerian isolate of banana streak badnavirus: I. Purification and enzyme-linked immunosorbent assay

G. Thottappilly, G. Dahal, B. E L Lockhart

Research output: Contribution to journalArticlepeer-review

44 Scopus citations

Abstract

A Nigerian isolate of banana streak badnavirus (BSV) was purified and a polyclonal antiserum was produced in mice. The antiserum titre was between 1:10 000 and 1:40 000 in enzyme-linked immunosorbent assay (ELISA), and showed a good specificity to BSV antigens. Comparative tests were carded out to determine the sensitivity and reliability of BSV antigen detection by double antibody sandwich (DAS)-ELISA, triple antibody sandwich (TAS)-ELISA, antigen coated plate (ACP)-ELISA, and protein-A coated antibody sandwich (PAS)-ELISA. TAS-ELISA using rabbit polyclonal antiserum to trap BSV and mouse polyclonal antiserum to detect the virus particles, was more sensitive than ACP-ELISA and PAS-ELISA and detected BSV in plant extracts from both symptomatic and some asymptomatic plants. However, immunosorbent electron microscopy detected more BSV-infected plants from asymptomatic plant samples than did TAS-ELISA. Results of this study showed that detection of BSV antigens in sap extracts by TAS-ELISA was most efficient with symptomatic tissues which occurred most frequently in the 'cool rainy' season. This suggests that for more reliable BSV-indexing of field samples, tissue sampling should be done during the rainy season when most BSV-infected plants express severe symptoms.

Original languageEnglish (US)
Pages (from-to)253-261
Number of pages9
JournalAnnals of Applied Biology
Volume132
Issue number2
DOIs
StatePublished - Apr 1998

Keywords

  • Banana streak badnavirus
  • Immunosorbent electron microscopy
  • Polyclonal antibody
  • TAS-ELISA
  • Virus detection

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