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Improvements in the Direct Fluorescent Antibody Technique for the Detection, Identification, and Quantification of Renibacterium salmoninarum in Salmonid Kidney Smears
Author(s) -
Cvitanich J. D.
Publication year - 1994
Publication title -
journal of aquatic animal health
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 52
eISSN - 1548-8667
pISSN - 0899-7659
DOI - 10.1577/1548-8667(1994)006<0001:iitdfa>2.3.co;2
Subject(s) - staining , fluorescence , biology , fluorescent staining , direct fluorescent antibody , immunofluorescence , kidney , microbiology and biotechnology , antibody , pathology , chromatography , chemistry , immunology , medicine , endocrinology , physics , quantum mechanics , genetics
Staining trials were conducted over a 5‐year period to explore ways of improving the direct fluorescent antibody technique for Renibacterium salmoninarum (Rs). In most salmonid kidney tissue smears that were pretreated with the organic solvent xylene, acetone, or methanol for 2 min, Rs had substantially higher fluorescence intensity and better uniformity of fluorescence than Rs in non‐pretreated controls. Only in smears prepared with cultured Rs cells were staining responses of Rs cells similar regardless of pretreatment. Increasing fluorescent antibody staining time greatly increased fluorescence intensity in most trials, but did not affect uniformity of fluorescence; 30 min was considered minimum or adequate staining time and 60 min was found to be optimal. Improved staining facilitated Rs detection and identification, allowing more reliable quantification in standardized smear preparations.