用于检测猪痢疾短螺旋体和结肠短螺旋体的培养、生化试验与聚合酶链反应的比较
Comparison of culture and biochemical tests with PCR for detection of Brachyspira hyodysenteriae and Brachyspira pilosicoli.
作者信息
Råsbäck T, Fellström C, Gunnarsson A, Aspán A
机构信息
Department of Clinical Sciences, Swedish University of Agricultural Sciences, P.O. Box 7018, SE-750 07 Uppsala, Sweden.
出版信息
J Microbiol Methods. 2006 Aug;66(2):347-53. doi: 10.1016/j.mimet.2005.12.008. Epub 2006 Feb 2.
Traditional culture and biochemical tests (CBT) were compared with PCR for sensitivity and detection of Brachyspira hyodysenteriae and Brachyspira pilosicoli in seeded faeces and clinical samples from diarrhoeic pigs. A duplex PCR system was developed based on primers detecting the tlyA-gene of B. hyodysenteriae and the 16S rRNA-gene of B. pilosicoli. Sensitivities for the PCR system were determined on seeded faeces, using DNA that had been recovered from primary cultures or extracted directly from faeces. Compared to CBT, PCR applied to DNA extracted directly from faeces lowered the sensitivity by a factor of 1000 to 10,000. B. hyodysenteriae and B. pilosicoli detection was compared for CBT and PCR using 200 clinical samples. CBT detected more B. hyodysenteriae isolates in the clinical samples than PCR, but fewer B. pilosicoli positive samples. An atypical strongly haemolytic isolate was detected only by CBT.
将传统文化和生化检测（CBT）与聚合酶链反应（PCR）相比较，以检测接种粪便以及腹泻猪临床样本中的猪痢疾短螺旋体和毛发状短螺旋体的敏感性。基于检测猪痢疾短螺旋体tlyA基因和毛发状短螺旋体16S rRNA基因的引物，开发了一种双重PCR系统。使用从原代培养物中回收或直接从粪便中提取的DNA，在接种粪便上测定PCR系统的敏感性。与CBT相比，应用于直接从粪便中提取的DNA的PCR将敏感性降低了1000至10000倍。使用200份临床样本比较了CBT和PCR对猪痢疾短螺旋体和毛发状短螺旋体的检测情况。CBT在临床样本中检测到的猪痢疾短螺旋体分离株比PCR多，但毛发状短螺旋体阳性样本较少。仅通过CBT检测到一株非典型强溶血分离株。
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