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Diseases of Aquatic Organisms

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DAO 36:45-51 (1999)  -  doi:10.3354/dao036045

Development of a polymerase chain reaction diagnostic assay for Ceratomyxa shasta, a myxosporean parasite of salmonid fish

Oswaldo Palenzuela1,*, Grant Trobridge2, Jerri L. Bartholomew1

1Department of Microbiology, Nash Hall, Room 220, Oregon State University, Corvallis, Oregon 97331-3804, USA
2Markey Molecular Medicine Center, Mailstop 357720, University of Washington, Seattle, Washington 98195-7720, USA

ABSTRACT: A diagnostic procedure based on the polymerase chain reaction (PCR) was developed for the myxosporean parasite Ceratomyxa shasta. Three sets of oligonucleotide primers were designed to specifically amplify C. shasta ribosomal RNA genes and several parameters of the assay were tested and optimised. A simple protocol for the processing of fish tissue samples was also developed. In a single round, 20 µl volume reaction the optimised procedure allows the detection of 50 fg of purified C. shasta genomic DNA, or 0.01 spore from a seeded fish intestine sample. This protocol is considerably faster, cheaper and more reliable than any previous diagnostic procedure for a myxosporean parasite, and can be an invaluable tool for the monitoring of early and/or subclinical C. shasta infections in wild and cultured salmon populations.

KEY WORDS: Myxozoa · Diagnostics · Ceratomyxa · Polymerase chain reaction · Ribosomal RNA gene · rDNA

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