DAO 71:149-154 (2006)  -  doi:10.3354/dao071149

Cost efficiency in the detection of chytridiomycosis using PCR assay

Kerry M. Kriger1,2,*, Jean-Marc Hero1, Kevin J. Ashton3

1Griffith University, Centre for Innovative Conservation Strategies, PMB 50 Gold Coast Mail Centre, Queensland 9726, Australia
2Save The Frogs, Centre for Advanced Scientific Research, 10995 Poplar Ford Trail, Manassas, Virginia 20109, USA
3Griffith University, Heart Foundation Research Centre, PMB 50 Gold Coast Mail Centre, Queensland 9726, Australia

ABSTRACT: Chytridiomycosis is an emerging infectious disease of amphibians associated with mass mortalities and population declines worldwide. Recent technological advances have resulted in a highly sensitive, non-invasive technique for diagnosing the disease based on a quantitative (real-time) polymerase chain reaction (qPCR) assay. The qPCR assay yields the most accurate and informative data of any available detection technique. However, due to the relatively high costs involved, it has yet to attain widespread use by chytridiomycosis researchers. Using the results of a disease survey of 467 wild frogs from eastern Queensland, Australia, we examine the necessity of triplicate assays in qPCR detection of chytridiomycosis. We describe a singlicate qPCR assay that can be used to substantially decrease costs, with no significant decrease in sensitivity. We also demonstrate that detection of chytridiomycosis by use of the conventional PCR assay may lead to appreciable underestimations in disease prevalence. We recommend that amphibian disease researchers adopt the singlicate qPCR assay as the primary means of chytridiomycosis detection.

KEY WORDS: Batrachochytrium dendrobatidis · Amphibian declines · Chytridiomycosis · Real-time Taqman PCR assay · Chytrid

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