A simplified PCR-based method for the detection of Renibacterium salmoninarum utilizing preparations of rainbow trout (Oncorhynchus mykiss, Walbaum) lymphocytes.
A method for the detection of Renibacterium salmoninarum by PCR is described. A rapid, reliable procedure was developed for the extraction of DNA, which could be applied to infected kidney homogenates and head kidney lymphocyte preparations. The target for DNA amplification was a 376-bp region of the gene encoding the 57-kDa major surface antigen (MSA). The PCR was specific for R. salmoninarum and allowed the detection of 10 to 100 cells of the pathogen. Use of the PCR for the examination of experimentally infected rainbow trout showed it to be as reliable as plate culture methods for the detection of R. salmoninarum in infected kidneys.
McIntosh
D
D
Department of Biological Sciences, Heriot-Watt University, Riccarton, Edinburgh, Scotland.
Meaden
P G
PG
Austin
B
B
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics; D000954:Antigens, Surface / Q000235:genetics; D001431:Bacteriological Techniques / Q000706:statistics & numerical data; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D005069:Evaluation Studies as Topic; D005393:Fish Diseases / Q000382:microbiology; D005798:Genes, Bacterial; D006094:Gram-Positive Bacteria / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D007674:Kidney Diseases / Q000382:microbiology / Q000662:veterinary; D008214:Lymphocytes / Q000382:microbiology; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000942:Antigens, Bacterial; D000954:Antigens, Surface; D017931:DNA Primers; D004269:DNA, Bacterial