Development of multiplex PCR assay for simultaneous detection of five bacterial fish pathogens

• Published in: Veterinary microbiology Vol. 131; no. 3; pp. 332 - 338
• Main Authors: Altinok, Ilhan, Capkin, Erol, Kayis, Sevki
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15-10-2008; Amsterdam; New York: Elsevier; Elsevier
• Subjects: accuracy; Aeromonas hydrophila; Aeromonas salmonicida subsp. salmonicida; Animals; Bacteria - isolation & purification; bacterial infections; Bacterial Infections - microbiology; Bacterial Infections - veterinary; Bacterial kidney disease; Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Columnaris; disease detection; disease diagnosis; Enteric red mouth disease; farmed fish; fish; fish diseases; Fish Diseases - microbiology; Flavobacterium columnare; Fundamental and applied biological sciences. Psychology; Furunculosis; kidney diseases; Microbiology; molecular epidemiology; molecular genetics; Motile Aeromonas septicemia; Multiplex PCR; multiplex polymerase chain reaction; Oncorhynchus mykiss; pathogen identification; polymerase chain reaction; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - veterinary; rapid methods; Renibacterium salmoninarum; Sensitivity and Specificity; septicemia; validity; Yersinia ruckeri; Multiplex PCR; Columnaris; Motile Aeromonas septicemia; Enteric red mouth disease; Bacterial kidney disease; Furunculosis; Kidney disease; Skin disease; Urinary system disease; Multiplex polymerase chain reaction; Septicemia; Aeromonas; Infection; Vertebrata; Pisces; Bacteriosis; Bacteria; Vibrionaceae; Detection; Veterinary
• ISSN: 0378-1135; 1873-2542
• DOI: 10.1016/j.vetmic.2008.04.014

A multiplex polymerase chain reaction (PCR) method was designed for the simultaneous detection of the five major fish pathogens,
Aeromonas hydrophila,
Aeromonas salmonicida subsp.
salmonicida,
Flavobacterium columnare,
Renibacterium salmoninarum, and
Yersinia ruckeri. Each of the five pairs of oligonucleotide primers exclusively amplified the targeted gene of the specific microorganism. The detection limits of the multiplex PCR was in the range of 2, 1, 1, 3, and 1
CFU for
A. hydrophila,
A. salmonicida,
F. columnare,
R. salmoninarum, and
Y. ruckeri, respectively. Multiplex PCR did not produce any nonspecific amplification products when tested against 23 related species of bacteria. The multiplex PCR assay was useful for the detection of the bacteria in naturally infected fish. This assay is a sensitive and specific and reproducible diagnostic tool for the simultaneous detection of five pathogenic bacteria that cause disease in fish. Therefore, it could be a useful alternative to the conventional culture based method.