Detection and quantification of Renibacterium salmoninarum DNA in salmonid tissues by real-time quantitative polymerase chain reaction analysis
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Abstract
Renibacterium salmoninarum is an important salmonid pathogen that is difficult to culture. We developed and assessed a real-time, quantitative, polymerase chain reaction (qPCR) assay for the detection and enumeration of R. salmoninarum. The qPCR is based on TaqMan technology and amplifies a 69-base pair (bp) region of the gene encoding the major soluble antigen (MSA) of R. salmoninarum. The qPCR assay consistently detected as few as 5 R. salmoninarum cells per reaction in kidney tissue. The specificity of the qPCR was confirmed by testing the DNA extracts from a panel of microorganisms that were either common fish pathogens or reported to cause false-positive reactions in the enzyme-linked immunosorbent assay (ELISA). Kidney samples from 38 juvenile Chinook salmon (Oncorhynchus tshawytscha) in a naturally infected population were examined by real-time qPCR, a nested PCR, and ELISA, and prevalences of R. salmoninarum detected were 71, 66, and 71%, respectively. The qPCR should be a valuable tool for evaluating the R. salmoninarum infection status of salmonids.
| Publication type | Article |
|---|---|
| Publication Subtype | Journal Article |
| Title | Detection and quantification of Renibacterium salmoninarum DNA in salmonid tissues by real-time quantitative polymerase chain reaction analysis |
| Series title | Journal of Veterinary Diagnostic Investigation |
| DOI | 10.1177/104063870601800409 |
| Volume | 18 |
| Issue | 4 |
| Year Published | 2006 |
| Language | English |
| Publisher | Sage Journals |
| Contributing office(s) | Western Fisheries Research Center |
| Description | 6 p. |
| First page | 375 |
| Last page | 380 |
| Online Only (Y/N) | N |
| Additional Online Files (Y/N) | N |
| Google Analytic Metrics | Metrics page |