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Diseases of Aquatic Organisms

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DAO 71:75-80 (2006)  -  doi:10.3354/dao071075

Development of a TaqMan PCR assay for the detection of Bonamia species

Serge Corbeil1,*, Isabelle Arzul2, Ben Diggles3, Mike Heasman4, Bruno Chollet2, Franck C. J. Berthe5, Mark St. J. Crane1

1AAHL Fish Diseases Laboratory, Australian Animal Health Laboratory, CSIRO Livestock Industries, Private Bag 24, Geelong, Victoria 3220, Australia
2IFREMER Laboratoire Génétique et Pathologie, Ronce-les-bains BP 133, 17390 La Tremblade, France
3DigsFish Services Pty. Ltd., Brisbane, Queensland, Australia
4NSW Fisheries, Port Stephens, New South Wales, Australia
5Department of Pathology & Microbiology, Atlantic Veterinary College, University of Prince Edward Island, Canada

ABSTRACT: The development of molecular diagnostic assays with increased sensitivity compared with conventional histological techniques is highly desirable for effective management of bonamiosis in cultured oyster stocks and wild populations. A real-time TaqMan PCR assay was developed for the specific detection of Bonamia species in infected oyster tissues. The TaqMan assay was shown to be significantly more sensitive than histopathology. Although a real-time TaqMan PCR assay is comparable with conventional PCR in terms of sensitivity, it offers the advantages that it is a rapid test and has a very low risk of sample cross-contamination. Furthermore, it can be optimised to quantify the parasite load in samples. The assay detected Bonamia isolates from Australia, New Zealand, Europe, Canada, Chile and the USA and therefore demonstrated genus specificity as tested in this study.

KEY WORDS: Bonamia spp. · Real-time TaqMan PCR assay · Oyster

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