Andree KB

El-Matbouli M

Lom J

Lorz HV

MacConnell E

Nehring RB

Pusterla N

Sollid SS

Vincent ER

Wagner EJ

 Diagnostic methods were used to identify and quantify Myxobolus cerebralis, a myxozoan parasite of salmonid fish. In this study, 7-week-old, pathogen-free rainbow trout ( Oncorhynchus mykiss) were experimentally infected with M. cerebralis and at 7 months postinfection were evaluated with 5 diagnostic assays: 1) pepsin–trypsin digest (PTD) to detect and enumerate spores found in cranial cartilage, 2) 2 different histopathology grading scales that provide a numerical score for severity of microscopic lesions in the head, 3) a conventional single-round polymerase chain reaction (PCR), 4) a nested PCR assay, and 5) a newly developed quantitative real-time TaqMan PCR. There were no significant differences ( P &gt; 0.05) among the 5 diagnostic assays in distinguishing between experimentally infected and uninfected control fish. The 2 histopathology grading scales were highly correlated ( P &lt; 0.001) for assessment of microscopic lesion severity. Quantification of parasite levels in cranial tissues using PTD and real-time TaqMan PCR was significantly correlated r = 0.540 ( P &lt; 0.001). Lastly, 104 copies of the 18S rDNA gene are present in the M. cerebralis genome, a feature that makes this gene an excellent target for PCR-based diagnostic assays. Also, 2 copies of the insulin growth factor–I gene are found in the rainbow trout genome, whose detection can serve both as an internal quality control for amplifiable DNA and as a basis to quantify pathogen genome equivalents present in quantitative PCR assays. </jats:p

Garry O. Kelley

Francisco J. Zagmutt-Vergara

Christian M. Leutenegger

Karin A. Myklebust

Mark A. Adkison

Terry S. McDowell

Gary D. Marty

Alex L. Kahler

Arla L. Bush

Ian A. Gardner

Ronald P. Hedrick

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Journal of Veterinary Diagnostic Investigation

Evaluation of Five Diagnostic Methods for the Detection and Quantification of <i>Myxobolus Cerebralis</i>

Evaluation of Five Diagnostic Methods for the Detection and Quantification of
                    Myxobolus Cerebralis

Evaluation of Five Diagnostic Methods for the Detection and Quantification of Myxobolus Cerebralis

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