DAO 49:185-190 (2002)  -  doi:10.3354/dao049185

Validation of a single round polymerase chain reaction assay for identification of Myxobolus cerebralis myxospores

Thomas J. Baldwin1,2,*, Karin A. Myklebust2

1Department of Veterinary Microbiology and Pathology, and
2Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, Washington 99164, USA
*Present address: Utah Veterinary Diagnostic Laboratory, 950 East, 1400 North Logan, Utah 84322-5700, USA. E-mail:

ABSTRACT: Validation of a single round PCR-based assay to confirm as Myxobolus cerebralis myxospores obtained from pepsin-trypsin digest preparations is described. The assay is a modification of a PCR assay published previously, based on the amplification of a segment of the gene encoding the 18S ribosomal subunit of M. cerebralis. The sensitivity, specificity and upper and lower detection limits were determined using known M. cerebralis and non-M. cerebralis myxospores and M. cerebralis-free fish. The sensitivity of PCR confirmation was 100% (95% confidence interval of 83.2-100%). The specificity was 100% (95% confidence interval of 87.2-100%). The upper detection limit was approximately 100000 myxospores per reaction; the lower detection limit was approximately 50 myxospores per reaction. Given the high sensitivity and specificity of the assay, substitution of this assay for histologic confirmation of M. cerebralis infection is encouraged.

KEY WORDS: Myxobolus cerebralis · Whirling disease

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