环介导等温扩增法快速检测感染性腹泻病例标本中的空肠弯曲菌

Rapid detection of Campylobacter jejuni in infectious diarrhea samples by loop-mediated isothermal amplification

中文关键词:  空肠弯曲菌  环介导等温扩增  滤膜法  感染性腹泻

英文关键词:Campylobacter jejuni  loop-mediated isothermal amplification  filter membrane method  infectious diarrhea

中文摘要:

      目的 建立环介导等温扩增法(loop-mediated isothermal amplification, LAMP)快速检测感染性腹泻病例标本中的空肠弯曲菌。方法 收集2017年苏州市辖区3家医院的感染性腹泻患者的粪便样品326件, 用LAMP法对空肠弯曲菌的DNA进行检测, 在DNA扩增试剂盒提供的反应体系加入引物和模板进行空肠弯曲菌的LAMP反应, 测定LAMP方法检测空肠弯曲菌菌液的灵敏度; 同时用滤膜法进行空肠弯曲菌的分离培养, 参照GB 4789.9-2014《食品安全国家标准 食品微生物学检验 空肠弯曲菌检验》分离鉴定空肠弯曲菌。结果 LAMP法检测空肠弯曲菌菌液的灵敏度是2.2 CFU/μL; 326件腹泻患者粪便中16件空肠弯曲菌LAMP检测为阳性, 阳性率为4.91%。共分离培养出14株空肠弯曲菌, 阳性率为4.29%。结论 针对感染性腹泻患者的粪便样本的空肠弯曲菌检测项目, 可以采用LAMP方法进行等温扩增初筛, 初筛阳性的样品再有针对性地进行后续的传统培养。

英文摘要:

      Objective To establish a rapid method for rapid detection of campylobacter jejuni (C. jejuni) in infectious diarrhea cases stool specimens by Loop-mediated isothermal amplification(LAMP). Methods Totally 326 stool specimens were collected from infectious diarrhea cases in 3 hospitals in Suzhou in 2017. The DNA of Campylobacter jejuni was detected by LAMP method. The LAMP reaction of Campylobacter jejuni was carried out by adding primers and template to the reaction system provided by DNA amplification kit. The sensitivity of LAMP method for detecting Campylobacter jejuni was determined. At the same time, the isolation and culture of Campylobacter jejuni were carried out by the filter method, and identified according to GB 4789.9-2014 Food safety national standard food microbiology test Campylobacter jejuni. Results The sensitivity of the LAMP method for detecting Campylobacter jejuni was 2.2 CFU/μL. Among the 326 patients with diarrhea, 16 cases of Campylobacter jejuni LAMP were positive, with a positive rate of 4.91%. A total of 14 strains of Campylobacter jejuni were isolated and cultured, and the positive rate was 4.29%. Conclusion For the detection of Campylobacter jejuni in stool samples from patients with infectious diarrhea, the LAMP method can be used for isothermal amplification primary screening, and the positive screening samples are targeted for subsequent traditional culture.