活体长爪沙鼠幽门螺杆菌检测方法的建立

发布时间：2018-03-04 07:35

本文选题：长爪沙鼠　切入点：幽门螺杆菌　出处：《中国比较医学杂志》2017年03期 　论文类型：期刊论文

【摘要】：目的应用巢式PCR方法通过胃液活体检测长爪沙鼠幽门螺杆菌感染,从而建立可以持续监测长爪沙鼠幽门螺杆菌感染的检测技术。方法体外培养幽门螺杆菌并接种至长爪沙鼠体内,在感染后第10周使用普通PCR方法检测长爪沙鼠胃液,使用巢式PCR方法对长爪沙鼠胃液、胃组织、十二指肠内容物、结肠粪便进行检测;同时还利用快速尿素酶试验和血清ELISA方法等对比分析巢式PCR方法的准确性,上述PCR产物均经过测序验证。结果普通PCR方法检测胃液、巢式PCR方法检测胃液、胃组织、十二指肠内容物,结肠粪便以及快速尿素酶试验、血清ELISA检测方法检验结果阳性率分别为30%、100%、100%、90%、10%、100%和0%。比较不同检测方法的检测结果发现,胃液巢式PCR、胃组织巢式PCR和快速尿素酶试验均有100%的检测率,普通PCR检测胃液的方法、结肠粪便巢式PCR方法及血清学ELISA检测方法检测率均比较低。结论胃液巢式PCR检测方法由于其特异性、准确性及灵敏性较高等特点可以用于长爪沙鼠幽门螺杆菌的长期检测,特别是对于预防和治疗幽门螺杆菌感染的实验具有重要价值。
[Abstract]:The purpose of a nested PCR method for in vivo detection of gastric juice by gerbils infected with Helicobacter pylori in Mongolian gerbils can continuously monitor the detection of Helicobacter pylori infection. In order to establish technology of Helicobacter pylori and inoculated into the gerbil in vivo culture method in vitro, in tenth weeks after infection with the common PCR method for detection of Mongolian gerbil gastric juice on Mongolian gerbil gastric juice, gastric tissues by nested PCR method, duodenal contents, colon feces were detected; and also the use of the rapid urease test and serum ELISA method comparative analysis the accuracy of the nested PCR method, the PCR products were verified by sequencing. Results the common PCR method for detection of gastric juice, the nested PCR detection of gastric juice, gastric tissue. Duodenal contents, fecal and rapid urease test, the test results of serum ELISA detection positive rate were 30%, 100%, 100%, 90%, 10%, 100% and 0%. Results detection method for comparison of different gastric juice, nested PCR, nested PCR detection of gastric tissue and rapid urease test were 100% rate, general methods of PCR detection of gastric juice, fecal nested PCR method and serological detection method for ELISA detection rate was relatively low. Conclusion gastric fluid detection method of nested PCR because of its specific characteristics higher accuracy and sensitivity can be used for long-term detection of Helicobacter pylori in Mongolian gerbils, especially has important value for the prevention and treatment of Helicobacter pylori infection in the experiment.

【作者单位】：首都医科大学基础医学院;中国食品药品检定研究院实验动物资源研究所;
【基金】：国家科技支撑计划课题(2015BAI09B01) 国家自然科学基金项目(31572348) 北京自然科学基金重点项目(714100)
【分类号】：R-332;R573

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