Lorch Jeffrey M, Gargas Andrea, Meteyer Carol Uphoff, Berlowski-Zier Brenda M, Green D Earl, Shearn-Bochsler Valerie, Thomas Nancy J, Blehert David S
U.S. Geological Survey-National Wildlife Health Center, 6006 Schroeder Road, Madison, WI 53711, USA.
J Vet Diagn Invest. 2010 Mar;22(2):224-30. doi: 10.1177/104063871002200208.
A newly developed polymerase chain reaction (PCR)-based method to rapidly and specifically detect Geomyces destructans on the wings of infected bats from small quantities (1-2 mg) of tissue is described in the current study (methods for culturing and isolating G. destructans from bat skin are also described). The lower limits of detection for PCR were 5 fg of purified fungal DNA or 100 conidia per 2 mg of wing tissue. By using histology as the standard, the PCR had a diagnostic specificity of 100% and a diagnostic sensitivity of 96%, whereas the diagnostic sensitivity of culture techniques was only 54%. The accuracy and fast turnaround time of PCR provides field biologists with valuable information on infection status more rapidly than traditional methods, and the small amount of tissue required for the test would allow diagnosis of white-nose syndrome in live animals.
本研究描述了一种新开发的基于聚合酶链反应(PCR)的方法,该方法可从少量(1 - 2毫克)组织中快速、特异性地检测受感染蝙蝠翅膀上的毁灭柱孢菌(同时还描述了从蝙蝠皮肤培养和分离毁灭柱孢菌的方法)。PCR的检测下限为每2毫克翅膀组织中5飞克纯化真菌DNA或100个分生孢子。以组织学为标准,PCR的诊断特异性为100%,诊断敏感性为96%,而培养技术的诊断敏感性仅为54%。PCR的准确性和快速周转时间比传统方法能更快地为野外生物学家提供有关感染状况的有价值信息,并且该检测所需的组织量少,能够对活体动物进行白鼻综合征的诊断。