DEVELOPMENT OF A DNA-PROBE FOR THE MYXOSPOREAN PARASITE CERATOMYXA-SHASTA, USING THE POLYMERASE CHAIN-REACTION WITH ARBITRARY PRIMERS

The arbitrarily primed polymerase chain reaction (PCR) was used to gen erate a DNA marker specific for the myxosporean parasite Ceratomyxa sh asta. The [P-32]-labeled marker hybridized to purified C. shasta DNA a nd to parasite DNA combined with salmonid DNA in a dot blot assay, dem onstrating its potential as a diagnostic tool. The amplified DNA segme nt was cloned and sequenced, and primers specific for the marker were designed. When these primers were used in a standard PCR assay, DNA wa s amplified from C. shasta and from infected fish tissues, but not fro m uninfected fish tissues or from 2 other myxosporean parasites. The s ensitivity of the PCR assay will permit detection of low levels of C. shasta from infected fish or oligochaetes and will be useful in defini ng the parasite's Life cycle as well as examining its impact on salmon id populations.