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Simultaneous detection of protozoa in the tissues of snakes by double in situ hybridization
Author(s) -
Richter Barbara,
Fragner Karin,
Weissenböck Herbert
Publication year - 2008
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20546
Subject(s) - digoxigenin , protozoa , biology , in situ hybridization , pathogen , in situ , dig , chromogenic , microbiology and biotechnology , chemistry , chromatography , genetics , gene expression , gene , organic chemistry
Different methods have been established for the simultaneous detection of different pathogens in tissue samples, each with certain advantages and disadvantages. Chromogenic in situ hybridization combines specific molecular pathogen detection with microscopic evaluation of pathogen quantity, morphology and distribution, as well as associated tissue damage. Furthermore, only a minimum of usually costly technical equipment is needed. The aim of our study was to detect two different protozoa simultaneously in tissue samples using exclusively digoxigenin (DIG)‐labeled probes and alkaline phosphatase‐coupled anti‐DIG‐antibodies and the chromogens Vector Red and NBT/BCIP with standard protocols. Gastrointestinal tissue samples from 15 snakes infected with either one or two protozoan species were investigated. All expected protozoa stained clearly dark purple or bright red, respectively, depending on the chromogen used. This technique can be used in pathogenicity studies of various pathogens in any kind of tissue. Microsc. Res. Tech., 2008. © 2007 Wiley‐Liss, Inc.