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Specificity of PCR and in situ hybridization assays designed for detection of Marteilia sydneyi and M. refringens

Published online by Cambridge University Press:  16 January 2003

S. N. KLEEMAN ,
F. LE ROUX ,
F. BERTHE  and
R. D. ADLARD
S. N. KLEEMAN
Affiliation:
Department of Microbiology and Parasitology, University of Queensland, Brisbane, QLD 4072, Australia
F. LE ROUX
Affiliation:
Laboratoire de Génétique et Pathologie, IFREMER, BP133 Ronce-Les-Bains, 17390 La Tremblade, France
F. BERTHE
Affiliation:
Laboratoire de Génétique et Pathologie, IFREMER, BP133 Ronce-Les-Bains, 17390 La Tremblade, France
R. D. ADLARD
Affiliation:
Protozoa Section, Queensland Museum, PO Box 3300, South Brisbane, QLD 4101, Australia
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Abstract

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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 ITS1 probe. The ITS probe is proposed for use as a confirmatory diagnostic tool for M. sydneyi.

Keywords

in situ hybridizationMarteilia refringensMarteilia sydneyiMarteilioides spp.PCR, species specificity
Type
Original Article
Information
Parasitology , Volume 125 , Issue 2 , August 2002 , pp. 131 - 141
Copyright
2002 Cambridge University Press