Indirect immunofluorescence test for plant pathogenic bacteria
Published: Thu, 27 Dec 2012, 16:09Last updated: Mon, 11 May 2015, 16:16
This standard describes how to perform an indirect immunofluorescence test (IF) for plant pathogenic bacteria. Approved in 2009-09. Instructions to perform the IF test are given in Appendix 1. A validated source of antiserum or antibodies to the target organism should be used. If validation data are not provided by the supplier of the reagent, then the laboratory should generate these. The titre of the antiserum - antibodies should be determined for each batch. The titre is defined as the highest dilution at which optimum reaction occurs when testing a suspension containing 10^5-10^6 cells per mL of a reference strain of the target organism and using an appropriate dilution of antibody conjugate labelled with a fluorescent dye (e.g. fluorescein or rhodamin) according to the manufacturer-s recommendations. The recommended reference strain should preferably be the homologous strain, this is the strain used to produce the antiserum- antibody. Crude polyclonal antisera should, where available, have a minimum titre of 2000. During routine testing, the antiserum - antibodies should be used at a working dilution (WD) close to or at the minimum titre.
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- Diagnostic protocol