Development of a TaqMan PCR assay for the detection of Bonamia species.
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.
Corbeil
Serge
S
AAHL Fish Diseases Laboratory, Australian Animal Health Laboratory, CSIRO Livestock Industries, Private Bag 24, Geelong, Victoria 3220, Australia. serge.corbeil@csiro.au
Arzul
Isabelle
I
Diggles
Ben
B
Heasman
Mike
M
Chollet
Bruno
B
Berthe
Franck C J
FC
Crane
Mark St J
MS
D000818:Animals; D017931:DNA Primers / Q000737:chemistry; D016054:DNA, Protozoan / Q000032:analysis* / Q000302:isolation & purification; D041602:Haplosporida / Q000235:genetics / Q000302:isolation & purification*; D049895:Ostrea / Q000469:parasitology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D017931:DNA Primers; D016054:DNA, Protozoan; D012337:RNA, Ribosomal, 18S