A comparison of sensitivity between the real-time detection of a TaqMan® assay for Batrachochytrium dendrobatidis and conventional detection

Garland, Stephen, Wood, John, and Skerratt, Lee F. (2011) A comparison of sensitivity between the real-time detection of a TaqMan® assay for Batrachochytrium dendrobatidis and conventional detection. Diseases of Aquatic Organisms, 94 (2). pp. 101-105.

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Abstract

A sensitive and quantitative TaqMan® assay for the causative agent of chytridiomycosis in amphibians (Batrachochytrium dendrobatidis) has been developed and is routinely used in ­diagnostic laboratories. We assessed whether the real time detection of the TaqMan® assay was as sensitive as the detection of the PCR product by agarose gel electrophoresis and ethidium bromide staining. We found, for practical purposes, that gel-based detection of the diagnostic fragment produced by means of the TaqMan® assay or by conventional PCR that used a different polymerase and reaction mix was as sensitive as the real-time detection of the TaqMan® assay. We recommend the qualified use of conventional PCR amplification combined with agarose gel electrophoresis and ethidium bromide staining for studies where only prevalence data are required, funding for equipment is limited or the acquisition of a real-time system is not cost effective.

Item ID: 17302
Item Type: Article (Refereed Research - C1)
Keywords: Batrachochytrium dendrobatidis, chytridiomycosis, TaqMan assay, conventional PCR
ISSN: 1616-1580
Date Deposited: 28 Jun 2011 01:51
FoR Codes: 07 AGRICULTURAL AND VETERINARY SCIENCES > 0707 Veterinary Sciences > 070703 Veterinary Diagnosis and Diagnostics @ 50%
06 BIOLOGICAL SCIENCES > 0605 Microbiology > 060502 Infectious Agents @ 50%
SEO Codes: 96 ENVIRONMENT > 9608 Flora, Fauna and Biodiversity > 960805 Flora, Fauna and Biodiversity at Regional or Larger Scales @ 100%
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