PCR-based assays for the fish pathogen Aeromonas salmonicida. II. Further evaluation and validation of three PCR primer sets with infected fish.
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.
Byers
Helen K
HK
Australian Animal Health Laboratory, CSIRO Livestock Industries, Geelong, VIC.
Cipriano
Rocco C
RC
Gudkovs
Nicholas
N
Cranel
Mark St J
MS
D000333:Aeromonas / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D015169:Colony Count, Microbial / Q000662:veterinary; D003429:Cross Reactions; D017931:DNA Primers; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D005393:Fish Diseases / Q000175:diagnosis* / Q000382:microbiology; D016905:Gram-Negative Bacterial Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D017686:Oncorhynchus mykiss; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012474:Salmon; D012487:Salmonidae; D012680:Sensitivity and Specificity
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial