James Madison University, Department of Biology, 951 Carrier Dr., MSC 7801, Harrisonburg, VA, USA; National Oceanic and Atmospheric Administration (NOAA), National Ocean Service (NOS), National Center for Coastal Ocean Science (NCCOS), Center for Coastal Environmental Health and Biomolecular Research (CCEHBR) Lab, Charleston, SC, USA.
National Oceanic and Atmospheric Administration (NOAA), National Ocean Service (NOS), National Center for Coastal Ocean Science (NCCOS), Center for Coastal Environmental Health and Biomolecular Research (CCEHBR) Lab, Charleston, SC, USA.
Mol Cell Probes. 2021 Apr;56:101695. doi: 10.1016/j.mcp.2021.101695. Epub 2021 Jan 13.
Researchers have developed multiple methods to characterize clinical and environmental strains of Vibrio vulnificus. The aim of our study was to use four assays to detect virulence factors in strains from infected patients and those from surface waters/sediments/oysters of South Carolina and the Gulf of Mexico. Vibrio vulnificus strains from clinical (n = 81) and environmental (n = 171) sources were tested using three real-time PCR methods designed to detect polymorphisms in the 16S rRNA, vcg and pilF genes and a phenotypic method, the ability to ferment D-mannitol. Although none of the tests correctly categorized all isolates, the differentiation between clinical and environmental isolates was similar for the pilF, vcgC/E and 16S rRNA assays, with sensitivities of 74.1-79.2% and specificities of 77.4-82.7%. The pilF and vcgC/E assays are comparable in efficacy to the widely used 16S rRNA method, while the D-mannitol fermentation test is less discriminatory (sensitivity = 77.8%, specificity = 61.4%). Overall percent agreement for the D-mannitol fermentation method was also lower (66.7%) than overall percent agreement for the 3 molecular assays (78.0%-80.2%). This study demonstrated, using a large, diverse group of Vibrio vulnificus isolates, that three assays could be used to distinguish most clinical vs environmental isolates; however, additional assays are needed to increase accuracy.
研究人员已经开发出多种方法来描述临床和环境来源的创伤弧菌菌株。我们的研究目的是使用四种检测方法来检测来自感染患者和南卡罗来纳州及墨西哥湾的地表水/沉积物/牡蛎中的菌株的毒力因子。使用三种实时 PCR 方法检测来自临床（n=81）和环境（n=171）来源的创伤弧菌菌株，这些方法旨在检测 16S rRNA、vcg 和 pilF 基因中的多态性以及表型方法，即发酵 D-甘露醇的能力。尽管没有一种检测方法能正确地对所有分离株进行分类，但 pilF、vcgC/E 和 16S rRNA 检测方法对临床和环境分离株的区分相似，其敏感性为 74.1-79.2%，特异性为 77.4-82.7%。pilF 和 vcgC/E 检测方法与广泛使用的 16S rRNA 方法的效果相当，而 D-甘露醇发酵试验的区分度较低（敏感性=77.8%，特异性=61.4%）。D-甘露醇发酵方法的总一致性（66.7%）也低于三种分子检测方法的总一致性（78.0%-80.2%）。这项研究使用大量多样化的创伤弧菌分离株表明，三种检测方法可以区分大多数临床与环境分离株；然而，需要额外的检测方法来提高准确性。
