Detection of Salmonella spp in fecal specimens by use of real-time polymerase chain reaction assay
Detection of Salmonella spp in fecal specimens by use of real-time polymerase chain reaction assay
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DOI：
10.2460/ajvr.2002.63.1265
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发表时间：
2002-09-01
影响因子：
1
通讯作者：
Gentry-Weeks, CR
中科院分区：
文献类型：
--
作者：
Kurowski, PB;Traub-Dargatz, JL;Gentry-Weeks, CR
目的：应用实时荧光定量聚合酶链式反应(PCR)技术，建立一种检测粪便标本中沙门氏菌的高灵敏度、高特异性的检测方法。实时荧光定量聚合酶链式反应检测沙门氏菌特异性DNA片段。结果：与299份临床粪便标本的细菌培养结果比较，SpaQ法检测沙门氏菌的相对敏感度为100%，特异度为98.2%。结论与临床相关性--建立了一种快速、灵敏、特异的沙门氏菌检测方法。这项技术在临床环境中将非常有价值，有助于避免或减轻沙门氏菌病在牛群或兽医医院患者中暴发所引起的并发症。
Objective-To use real-time polymerase chain reaction (PCR) technology to develop a highly sensitive and specific diagnostic assay for the detection of Salmonella spp in fecal specimens.Sample Population-299 fecal specimens from cattle, horses, and dogs.Procedure-Enrichment of fecal specimens was followed by genomic DNA extraction by use of commercially available isolation kits. Real-time PCR assay was performed to target a Salmonella spp-specific DNA segment. Results of real-time PCR assay were compared with bacterial culture results to determine relative sensitivity and specificity.Results-Use of the spaQ primer-probe set resulted in a relative sensitivity of 100% and a specificity of 98.2%, compared with bacterial culture results when tested on 299 clinical fecal specimens.Conclusions and Clinical Relevance-A rapid, sensitive, and specific assay for the detection of Salmonella spp from enriched clinical fecal specimens was developed. This technique would be highly valuable in clinical settings to help avoid or mitigate the complications arising from an outbreak of salmonellosis in a herd or among patients of a veterinary hospital.