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Diseases of Aquatic Organisms

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DAO 77:11-15 (2007)  -  DOI:

Quantitative PCR detection of Batrachochytrium dendrobatidis DNA from sediments and water

Julie D. Kirshtein1, Chauncey W. Anderson2,*, John S. Wood3, Joyce E. Longcore4, Mary A. Voytek1

1US Geological Survey, MS 430, 12201 Sunrise Valley Dr., Reston, Virginia 20192, USA
2US Geological Survey, Oregon Water Science Center, 2130 SW Fifth Avenue, Portland, Oregon 97201, USA
3Pisces Molecular, 2200 Central Avenue, Suite F, Boulder, Colorado 80301, USA
4University of Maine, Department of Biological Sciences, 5722 Deering Hall, Orono, Maine 04469, USA
*Corresponding author. Email:

ABSTRACT: The fungal pathogen Batrachochytrium dendrobatidis (Bd) causes chytridiomycosis, a disease implicated in amphibian declines on 5 continents. Polymerase chain reaction (PCR) primer sets exist with which amphibians can be tested for this disease, and advances in sampling techniques allow non-invasive testing of animals. We developed filtering and PCR based quantitative methods by modifying existing PCR assays to detect Bd DNA in water and sediments, without the need for testing amphibians; we tested the methods at 4 field sites. The SYBR based assay using Boyle primers (SYBR/Boyle assay) and the Taqman based assay using Wood primers performed similarly with samples generated in the laboratory (Bd spiked filters), but the SYBR/Boyle assay detected Bd DNA in more field samples. We detected Bd DNA in water from 3 of 4 sites tested, including one pond historically negative for chytridiomycosis. Zoospore equivalents in sampled water ranged from 19 to 454 l–1 (nominal detection limit is 10 DNA copies, or about 0.06 zoospore). We did not detect DNA of Bd from sediments collected at any sites. Our filtering and amplification methods provide a new tool to investigate critical aspects of Bd in the environment.

KEY WORDS: Amphibian · Chytridiomycosis · Detection · Disease · Filter · PCR · Primers · Water

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Cite this article as: Kirshtein JD, Anderson CW, Wood JS, Longcore JE, Voytek MA (2007) Quantitative PCR detection of Batrachochytrium dendrobatidis DNA from sediments and water. Dis Aquat Org 77:11-15.

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