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


Altinok I., ÇAPKIN E., Kayis S.

VETERINARY MICROBIOLOGY, cilt.131, ss.332-338, 2008 (SCI İndekslerine Giren Dergi) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 131
  • Basım Tarihi: 2008
  • Doi Numarası: 10.1016/j.vetmic.2008.04.014
  • Dergi Adı: VETERINARY MICROBIOLOGY
  • Sayfa Sayıları: ss.332-338

Özet

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. (C) 2008 Elsevier B.V. All rights reserved.