Development of polymerase chain reaction assays for detection, identification, and differentiation of Piscirickettsia salmonis

• Published in: Diseases of aquatic organisms Vol. 26; no. 3; pp. 189 - 195
• Main Authors: MAUEL, M. J, GIOVANNONI, S. J, FRYER, J. L
• Format: Journal Article
• Language: English
• Published: Oldendorf Inter-Research 12-09-1996
• Subjects: Animal bacterial diseases; Bacterial diseases; Biological and medical sciences; Infectious diseases; Marine; Medical sciences; Piscirickettsia salmonis; Salmonidae; Salmonidae; Rickettsialosis; Infection; Rickettsiales; Polymerase chain reaction; Vertebrata; Restriction fragment length polymorphism; Rickettsial infection; DNA; Pathogenic; Pisces; Pisciculture; Bacteriosis; Bacteria; Rickettsieae; Diagnosis; Detection; Rickettsiaceae
• ISSN: 0177-5103; 1616-1580
• DOI: 10.3354/dao026189

A nested polymerase chain reaction (PCR) was developed to detect genomic DNA of Piscirickettsia salmonis, the causative agent of an epizootic disease in salmonids. The nested PCR assay, which used general bacterial 16S rDNA primers in the first amplification reaction, and P. salmonis-specific primers in a second reaction, allowed detection of less than 1 P. salmonis tissue culture infectious dose 50 (TCID sub(50)). Using the P. salmonis-specific primers in a single PCR reaction allowed the detection of 60 TCID sub(50). The specificity of the PCR was assessed with a panel of 4 salmonid and 15 bacterial genomic DNA preparations. Amplification products were produced only with P. salmonis DNA. Restriction fragment length polymorphism (RFLP) analysis of the complete 16S gene PCR products demonstrated that 1 isolate, EM-90, was unique. Two additional primers were developed and used in PCR assays that differentiated EM-90 from the 4 other P. salmonis isolates tested.