Bugarel M, Tudor A, Loneragan G H, Nightingale K K
International Center for Food Industry Excellence, Department of Animal and Food Sciences, Texas Tech University, Lubbock, TX, USA.
International Center for Food Industry Excellence, Department of Animal and Food Sciences, Texas Tech University, Lubbock, TX, USA.
J Microbiol Methods. 2017 Mar;134:14-20. doi: 10.1016/j.mimet.2016.12.011. Epub 2016 Dec 18.
Foodborne illnesses due to Salmonella represent an important public-health concern worldwide. In the United States, a majority of Salmonella infections are associated with a small number of serotypes. Furthermore, some serotypes that are overrepresented among human disease are also associated with multi-drug resistance phenotypes. Rapid detection of serotypes of public-health concern might help reduce the burden of salmonellosis cases and limit exposure to multi-drug resistant Salmonella. We developed a two-step real-time PCR-based rapid method for the identification and detection of five Salmonella serotypes that are either overrepresented in human disease or frequently associated with multi-drug resistance, including serotypes Enteritidis, Typhimurium, Newport, Hadar, and Heidelberg. Two sets of four markers were developed to detect and differentiate the five serotypes. The first set of markers was developed as a screening step to detect the five serotypes; whereas, the second set was used to further distinguish serotypes Heidelberg, Newport and Hadar. The utilization of these markers on a two-step investigation strategy provides a diagnostic specificity of 97% for the detection of Typhimurium, Enteritidis, Heidelberg, Infantis, Newport and Hadar. The diagnostic sensitivity of the detection makers is >96%. The availability of this two-step rapid method will facilitate specific detection of Salmonella serotypes that contribute to a significant proportion of human disease and carry antimicrobial resistance.
由沙门氏菌引起的食源性疾病是全球重要的公共卫生问题。在美国，大多数沙门氏菌感染与少数血清型有关。此外，在人类疾病中占比过高的一些血清型也与多重耐药表型有关。快速检测引起公共卫生关注的血清型可能有助于减轻沙门氏菌病病例的负担，并限制接触多重耐药沙门氏菌。我们开发了一种基于两步实时PCR的快速方法，用于鉴定和检测在人类疾病中占比过高或经常与多重耐药相关的五种沙门氏菌血清型，包括肠炎血清型、鼠伤寒血清型、纽波特血清型、哈达尔血清型和海德堡血清型。开发了两组四个标记物来检测和区分这五种血清型。第一组标记物作为检测这五种血清型的筛选步骤；而第二组用于进一步区分海德堡血清型、纽波特血清型和哈达尔血清型。在两步调查策略中使用这些标记物，对鼠伤寒血清型、肠炎血清型、海德堡血清型、婴儿沙门氏菌血清型、纽波特血清型和哈达尔血清型的检测诊断特异性为97%。检测标记物的诊断敏感性>96%。这种两步快速方法的可用性将有助于特异性检测导致很大比例人类疾病并携带抗菌耐药性的沙门氏菌血清型。
