PCR-based assays for the fish pathogen Aeromonas salmonicida. II. Further evaluation and validation of three PCR primer sets with infected fish

Dis Aquat Organ. 2002 May 10;49(2):139-44. doi: 10.3354/dao049139.

Abstract

Two Aeromonas salmonicida-specific polymerase chain reaction (PCR) tests and 1 A. salmonicida subsp. salmonicida-specific PCR test were used to screen salmonid populations that were either overtly or covertly infected with A. salmonicida subsp. salmonicida. It was demonstrated that these PCR assays could be used to replace the biochemical testing currently employed to confirm the identity of A. salmonicida isolates cultured from infected fish. The AP and PAAS PCR assays were also capable of direct detection of A. salmonicida in overtly infected fish, with mucus, gill and kidney samples most likely to yield a positive result. Culture was a more reliable method for the direct detection of A. salmonicida in covertly infected salmonids than was the direct PCR testing of tissue samples, with the AP and PAAS PCRs having a lower detection limit (LDL) of approximately 4 x 10(5) colony-forming units (CFU) g(-1) sample.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aeromonas / genetics
  • Aeromonas / growth & development
  • Aeromonas / isolation & purification*
  • Animals
  • Bacterial Proteins / genetics
  • Base Sequence
  • Colony Count, Microbial / veterinary
  • Cross Reactions
  • DNA Primers*
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / isolation & purification
  • Fish Diseases / diagnosis*
  • Fish Diseases / microbiology
  • Gram-Negative Bacterial Infections / diagnosis
  • Gram-Negative Bacterial Infections / microbiology
  • Gram-Negative Bacterial Infections / veterinary*
  • Oncorhynchus mykiss
  • Polymerase Chain Reaction / methods
  • Polymerase Chain Reaction / veterinary*
  • Salmon
  • Salmonidae*
  • Sensitivity and Specificity

Substances

  • Bacterial Proteins
  • DNA Primers
  • DNA, Bacterial