实时定量聚合酶链反应结合分子信标与巢式聚合酶链反应及培养方法检测牛粪便样本中副结核分枝杆菌的比较
Comparison of real-time, quantitative PCR with molecular beacons to nested PCR and culture methods for detection of Mycobacterium avium subsp. paratuberculosis in bovine fecal samples.
作者信息
Fang Ying, Wu Wai-Hong, Pepper Jessica L, Larsen Jill L, Marras Salvatore A E, Nelson Eric A, Epperson William B, Christopher-Hennings Jane
机构信息
Animal Disease Research and Diagnostic Laboratory, South Dakota State University, Brookings, South Dakota 57007, USA.
出版信息
J Clin Microbiol. 2002 Jan;40(1):287-91. doi: 10.1128/JCM.40.1.287-291.2002.
Abstract
An automated PCR with fluorescent probes (molecular beacons) detected Mycobacterium avium subsp. paratuberculosis in bovine feces. When the PCR was compared with culture in testing 41 fecal samples, kappa scores of 0.94 to 0.96, a sensitivity of 93 to 96%, and a specificity of 92% were obtained. Results were quantitated by using a standard curve derived from a plasmid containing IS900. A minimum quantity of 1.7 x 10(-4) pg of DNA, correlating to 1 to 8 CFU, was detected.
摘要
一种带有荧光探针（分子信标）的自动化聚合酶链反应（PCR）检测出牛粪便中的副结核分枝杆菌。在检测41份粪便样本时，将该PCR与培养法进行比较，得到的kappa值为0.94至0.96，灵敏度为93%至96%，特异性为92%。通过使用源自含有IS900的质粒的标准曲线对结果进行定量。检测到的DNA最小量为1.7×10⁻⁴ pg，相当于1至8个菌落形成单位（CFU）。
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