Li Ruichao, Chiou Jiachi, Chan Edward Wai-Chi, Chen Sheng
Shenzhen Key lab for Food Biological Safety Control, Food Safety and Technology Research Center, Hong Kong PolyU Shen Zhen Research InstituteShenzhen, China; State Key Laboratory of Chirosciences, Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic UniversityHung Hom, Hong Kong.
Front Microbiol. 2016 Jan 28;7:44. doi: 10.3389/fmicb.2016.00044. eCollection 2016.
A PCR-based assay was developed for more accurate identification of Vibrio parahaemolyticus through targeting the bla CARB-17 like element, an intrinsic β-lactamase gene that may also be regarded as a novel species-specific genetic marker of this organism. Homologous analysis showed that bla CARB-17 like genes were more conservative than the tlh, toxR and atpA genes, the genetic markers commonly used as detection targets in identification of V. parahaemolyticus. Our data showed that this bla CARB-17-specific PCR-based detection approach consistently achieved 100% specificity, whereas PCR targeting the tlh and atpA genes occasionally produced false positive results. Furthermore, a positive result of this test is consistently associated with an intrinsic ampicillin resistance phenotype of the test organism, presumably conferred by the products of bla CARB-17 like genes. We envision that combined analysis of the unique genetic and phenotypic characteristics conferred by bla CARB-17 shall further enhance the detection specificity of this novel yet easy-to-use detection approach to a level superior to the conventional methods used in V. parahaemolyticus detection and identification.
开发了一种基于聚合酶链反应（PCR）的检测方法，通过靶向bla CARB-17样元件来更准确地鉴定副溶血性弧菌，该元件是一种内在的β-内酰胺酶基因，也可被视为该生物体的一种新型物种特异性遗传标记。同源性分析表明，bla CARB-17样基因比tlh、toxR和atpA基因更保守，后三者是副溶血性弧菌鉴定中常用作检测靶点的遗传标记。我们的数据表明，这种基于bla CARB-17特异性PCR的检测方法始终具有100%的特异性，而靶向tlh和atpA基因的PCR偶尔会产生假阳性结果。此外，该检测的阳性结果始终与受试生物体的内在氨苄西林抗性表型相关，推测是由bla CARB-17样基因的产物赋予的。我们设想，对bla CARB-17赋予的独特遗传和表型特征进行联合分析，将进一步提高这种新颖且易于使用的检测方法的检测特异性，使其优于副溶血性弧菌检测和鉴定中使用的传统方法。
