Specificity of PCR and in situ hybridization assays designed for detection of Marteilia sydneyi and M. refringens.
Primers and DNA probes designed for use in the specific detection of the paramyxean parasites Marteilia sydneyi and Marteilia refringens were tested for their potential to cross-react with closely related species in Polymerase Chain Reaction (PCR) and in situ hybridization. PCR primers and a DNA probe designed within the ITS1 rRNA of M. sydneyi were specific for M. sydneyi when compared with related species of Marteilia and Marteilioides. PCR primers designed within the 18S rRNA of M. refringens were specific in the detection of this species in PCR while a DNA probe (named Smart 2) designed on the same gene cross-reacted with M. sydneyi in tissue sections of Saccostrea glomerata as well as Marteilioides sp. infecting Striostrea mytiloides. Though not species specific, the Smart 2 probe provided a stronger signal in detection of all stages of M. sydneyi than the ITSI probe. The ITS probe is proposed for use as a confirmatory diagnostic tool for M. sydneyi.
Kleeman
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Department of Microbiology and Parasitology, University of Queensland, Brisbane, Australia. Sarah.Kleeman@affa.gov.au
Le Roux
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Berthe
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Adlard
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D000818:Animals; D017403:In Situ Hybridization / Q000379:methods*; D010125:Ostreidae / Q000166:cytology / Q000469:parasitology*; D010271:Parasites / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity