Detection Protocols - Renibacterium salmoninarum

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By Fish Health Program (FHP) September 4, 2018

Quantitative Culture for R. salmoninarum

Nested PCR for R. salmoninarum DNA

Quantitative PCR for R. salmoninarum DNA

Protocols for the Detection of Renibacterium salmoninarum in salmon

Bacterial Kidney Disease (BKD) in a juvenile salmon. — Kidney lesions in a juvenile Chinook salmon (Oncorhynchus tshawytscha) affected by bacterial kidney disease (BKD). Credit: Caroline Bentley, PhD., University of Central Oklahoma. (Public domain.)

Renibacterium salmoninarum is a fastidious, gram-positive bacterium and the causative agent of bacterial kidney disease (BKD) in wild and cultured salmonid fishes. A number of diagnostic tests exist for the direct and indirect detection of R. salmoninarum. These tests have different analytical properties and different abilities to quantify infection levels, detect remote infections and distinguish between live and dead bacteria. This website links to standard operating procedures (SOP) for the major R. salmoninarum diagnostic tests.

Quantitative Culture for Rs Image 1Thumb — Renibacterium salmoninarum colonies growing on a spread plate inoculated for quantitative culture of the bacterium from fish tissues or ovarian fluid. (Public domain.)

Quantitative Culture for R. salmoninarum - (2011) (Culture of Renibacterium salmoninarum from Tissues and Ovarian Fluid)

ELISA for Renibacterium salmoninarum Thumb — Double polyclonal enzyme-linked immunosorbent assay (ELISA) for the detection of Renibacterium salmonarium. Portion of a microplate showing green coloration in wells in which tissue samples containing high levels of R. salmoninarum antigens had been placed. (Public domain.)

Enzyme linked immunosorbent assay (ELISA) for R. salmoninarum – (2012) (Double Polyclonal Enzyme-Linked Immunosorbent Assay (ELISA) for the detection of Renibacterium salmoninarum)

DFAT for Renibacterium salmonarum Thumb — Direct fluorescent antibody test (DFAT) for the detection of Renibacterium salmoninarum in tissues. Fluorescing R. salmoninarum cells are visible on a slide. Credit: Carla Moanikeala Conway , USGS (Public domain.)

Direct fluorescent antibody test (DFAT) – (2011) (Direct Fluorescent Antibody Test (DFAT) for the detection of Renibacterium salmoninarum in tissues)

MF-FAT for Renibacterium salmoninarum Thumb — Membrane-filtration fluorescent antibody test (MF-FAT) for the detection of Renibacterium salmoninarum. A sample is loaded onto a membrane filter held in a syringe-mounted holder. (Public domain.)

Membrane filtration fluorescent antibody test (MF-FAT) – (2012) (Membrane-Filtration Fluorescent Antibody Test (MF-FAT) for the detection of Renibacterium salmoninarum)

DNA extraction of Renibacterium salmoninarum Thumb — DNA extraction for PCR-based detection of Renibacterium salmoninarum requires careful handling of samples to avoid contamination. (Public domain.)

DNA extraction for gram positive bacteria – (2011) (DNA Extraction for PCR-based detection of Renibacterium salmoninarum)

Photo of scientist at loading gel Thumb — Nested polymerase chain reaction (nPCR) for the detection of Renibacterium salmoninarum. Agarose gel electrophoresis is used for size separation and visualization of amplified DNA sequences. Credit: Debra Becker, USGS. (Public domain.)

Nested PCR (nPCR) test – (2011) (Nested Polymerase Chain Reaction (nPCR) for the detection of Renibacterium salmoninarum)

qPCR for detection of Rs 2Thumb — Quantitative polymerase chain reaction (qPCR) assay for the detection of Renibacterium salmoninarum DNA. Observation of qPCR results on the screen of a sequence detection system. Credit: Debra Becker, USGS. (Public domain.)

Quantitative PCR (qPCR) test – (Quantitative Polymerase Chain Reaction (qPCR) for the detection of Renibacterium salmoninarum)

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