College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, USA.
Lett Appl Microbiol. 2010 Feb;50(2):153-7. doi: 10.1111/j.1472-765X.2009.02770.x. Epub 2009 Nov 6.
To verify the specificity of a PCR assay for the identification and diagnosis of Edwardsiella ictaluri.
An Edwardsiella ictaluri-specific PCR assay was developed utilizing two features of the ribosomal DNA gene clusters. The first feature is the presence of two ribosomal gene clusters located in tandem to one another (the inter-ribosomal spacer, IRS). This characteristic is present in the Edwardsiella genus but absent in the other sequenced members of the Enterobacteriaceae. The second feature is the presence of an intervening sequence (IVS) in the 23S rRNA gene of Edw. ictaluri. To verify the specificity of this assay, we tested genomic DNA from a variety of bacterial species. The IVS/IRS PCR assay results in an c. 2000-bp product from all Edw. ictaluri isolates tested, but not from any other species including Edwardsiella tarda.
The IVS/IRS PCR assay is highly specific for Edw. ictaluri and useful as a tool for identifying this pathogen.
This research verifies the specificity of PCR-based assay for Edw. Ictaluri, and we describe this assay as a highly versatile diagnostic tool for its identification.
验证用于鉴定和诊断爱德华氏菌的 PCR 检测方法的特异性。
利用核糖体 DNA 基因簇的两个特征，开发了一种针对爱德华氏菌的特异性 PCR 检测方法。第一个特征是存在两个彼此串联排列的核糖体基因簇（核糖体间隔区，IRS）。这种特征存在于爱德华氏菌属中，但在肠杆菌科的其他已测序成员中不存在。第二个特征是 Edw. ictaluri 的 23S rRNA 基因中存在内含子（IVS）。为了验证该检测方法的特异性，我们测试了来自各种细菌物种的基因组 DNA。IVS/IRS PCR 检测方法会从所有测试的 Edw. ictaluri 分离株中产生约 2000bp 的产物，但不会从任何其他物种（包括迟缓爱德华氏菌）中产生。
IVS/IRS PCR 检测方法对 Edw. ictaluri 具有高度特异性，可用作鉴定该病原体的工具。
本研究验证了基于 PCR 的检测方法对爱德华氏菌的特异性，并将该检测方法描述为一种非常通用的鉴定工具。
