一种用于检测粪便和组织样本中副结核分枝杆菌鸟亚种的DNA序列捕获提取方法。
A DNA sequence capture extraction method for detection of Mycobacterium avium subspecies paratuberculosis in feces and tissue samples.
作者信息
Vansnick Elke, de Rijk Pim, Vercammen Francis, Rigouts Leen, Portaels Françoise, Geysen Dirk
机构信息
Institute of Tropical Medicine, Department of Animal Health, Nationalestraat 155, 2000 Antwerp, Belgium.
出版信息
Vet Microbiol. 2007 May 16;122(1-2):166-71. doi: 10.1016/j.vetmic.2007.01.011. Epub 2007 Jan 20.
Culturing of Mycobacterium avium subspecies paratuberculosis (Map) remains difficult and is time consuming. An alternative for the rapid detection of Map in samples is PCR. We have developed a sensitive DNA-extraction method based on sequence capture for the rapid detection of M. avium subspecies paratuberculosis by PCR in fecal and tissue samples. The method detected 10(2)Map/g feces using spiked samples, and reached a diagnostic sensitivity of 33,7% compared to 22% for culture. Analysis of tissue samples gave 65 polymerase chain reaction (PCR)-positive (42.2%) and 49 culture-positive samples (31.8%). Therefore, the detection limit of the DNA-extraction is the same as previously reported for culture, the PCR assay could detect more positive samples than the culture method.
副结核分枝杆菌（Map）的培养仍然困难且耗时。样本中Map快速检测的一种替代方法是聚合酶链反应（PCR）。我们开发了一种基于序列捕获的灵敏DNA提取方法，用于通过PCR在粪便和组织样本中快速检测副结核分枝杆菌。该方法使用加标样本检测到每克粪便含10²个Map，与培养法相比，诊断灵敏度达到33.7%，而培养法为22%。组织样本分析得到65个聚合酶链反应（PCR）阳性样本（42.2%）和49个培养阳性样本（31.8%）。因此，DNA提取的检测限与之前报道的培养法相同，PCR检测法能检测到比培养法更多的阳性样本。
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