Comparison of sensitivity between real-time ­detection of a TaqMan assay for Batrachochytrium dendrobatidis and conventional detection

Diseases of Aquatic Organisms

John Wood¹  Lee F. Skerratt¹  Stephen Garland¹ 

关键词: Batrachochytrium dendrobatidis;    Chytridiomycosis;    TaqMan assay;    Conventional PCR;   

DOI  :  10.3354/dao02327

学科分类：生物科学（综合）

来源: Inter-Research

【 摘 要 】

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.

【 授权许可】

Unknown