Development of a dot-immunobinding assay for detection of citrus tristeza virus.

Abstract:

:The dot-immunobinding assay (DIBA) was adapted for detection of citrus tristeza virus (CTV) and compared with DAS-ELISA and DAS-indirect ELISA. DIBA was easy to perform and as sensitive as either ELISA procedure for CTV diagnosis. The entire test could be performed in 2-3 h using polyclonal antibodies, with minimal laboratory equipment. Three different polyclonal antibodies gave a strong positive reaction with 12 selected CTV isolates; however, each serum had to be cross-absorbed with sap from healthy plants before use. The broad spectrum 3DF1 monoclonal antibody reacted with most of the CTV isolates. The MCA-13 strain-specific monoclonal antibody was specific for most severe CTV isolates. As blocking agents, 3% bovine serum albumin (BSA), 3% gelatin, 0.5% non-fat dry milk or 5% Triton X-100 gave an adequate white background on the nitrocellulose membranes and permitted discrimination between infected and healthy samples. However, 3% gelatin gave the best contrast between green for the healthy samples, and purple color for infected samples.

journal_name

J Virol Methods

authors

Rocha-Peña MA,Lee RF,Niblett CL

doi

10.1016/0166-0934(91)90108-c

subject

Has Abstract

pub_date

1991-10-01 00:00:00

pages

297-309

issue

3

eissn

0166-0934

issn

1879-0984

pii

0166-0934(91)90108-C

journal_volume

34

pub_type

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