国家自然科学基金项目（面上项目，重点项目，重大项目）;国家科技攻关计划
The National Natural Science Foundation of China (General Program, Key Program, Major Research Plan);The National Key Technologies R&D Program of China
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：体外培养幽门螺旋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旋杆菌的长期检测，特别是对于预防和治疗幽门螺旋杆菌感染的实验具有重要价值。
Objective: To detect H.pylori infection in living Mongolian gerbils using gastric juice and nested PCR, in order to establish a method to continuously detect H.pyloriinfection in Mongolian gerbils. Methods: Mongolian gerbils were infected with H. pylori which were cultured in vitro. The H.pylori weas detected with normal PCR through gastric juice and nested PCR through gastric juice, gastric mucosa, contents in duodenum and stool in colon. And use rapid urea test and ELISA to analyze the accuracy of the nested PCR method. All of the PCR products were verified by sequencing. Results：The positive test rates of normal PCR method through gastric juice, nested PCR method through gastric juice, gastric mucosa, contents in duodenum and stool in colon, rapid urea test and ELISA were 30%, 100%, 100%, 90%, 10%, 100% and 0%. Comparison of test results in different methods we found that the positive test rates are 100% in nested PCR through gastric juice; gastric mucosa and rapid urea test, but normal PCR method through gastric juice, nested PCR method through stool in colon, and ELISA were lower than other methods. Conclusion：This method can be used to detect H.pylori infection in Mongolian gerbils because of its specificity, accuracy and sensitivity, and it had an important value especially in prevention and treatment of H. pylori infection experiments.
王存龙,李长龙,邢进,冯育芳,杜小燕,岳秉飞,贺争鸣,陈振文.活体长爪沙鼠幽门螺旋杆菌检测方法的建立[J].中国比较医学杂志,2017,27().
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