Bat white-nose syndrome: a real-time TaqMan polymerase chain reaction test targeting the intergenic spacer region of Geomyces destructans

被引:123
作者
Muller, Laura K. [1 ]
Lorch, Jeffrey M. [2 ]
Lindner, Daniel L. [3 ]
O'Connor, Michael [4 ]
Gargas, Andrea [5 ]
Blehert, David S. [1 ]
机构
[1] US Geol Survey, Natl Wildlife Hlth Ctr, Madison, WI 53711 USA
[2] Univ Wisconsin, Med Sci Ctr, Mol & Environm Toxicol Ctr, Madison, WI 53706 USA
[3] US Forest Serv, No Res Stn, Ctr Forest Mycol Res, Madison, WI 53726 USA
[4] Wisconsin Vet Diagnost Lab, Madison, WI 53706 USA
[5] Symbiol LLC, Middleton, WI 53562 USA
关键词
diagnostic test; IGS; pathogen; PCR; wildlife disease; PCR AMPLIFICATION; FUNGAL PATHOGEN; RIBOSOMAL-DNA; SEQUENCES; INFECTION;
D O I
10.3852/12-242
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The fungus Geomyces destructans is the causative agent of white-nose syndrome (WNS), a disease that has killed millions of North American hibernating bats. We describe a real-time TaqMan PCR test that detects DNA from G. destructans by targeting a portion of the multicopy intergenic spacer region of the rRNA gene complex. The test is highly sensitive, consistently detecting as little as 3.3 fg genomic DNA from G. destructans. The real-time PCR test specifically amplified genomic DNA from G. destructans but did not amplify target sequence from 54 closely related fungal isolates (including 43 Geomyces spp. isolates) associated with bats. The test was qualified further by analyzing DNA extracted from 91 bat wing skin samples, and PCR results matched histopathology findings. These data indicate the real-time TaqMan PCR method described herein is a sensitive, specific and rapid test to detect DNA from G. destructans and provides a valuable tool for WNS diagnostics and research.
引用
收藏
页码:253 / 259
页数:7
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