Tamatsukuri S, Weiss J, Hirota K, Kawamata O, Shiratori Y, Yoshida H, Omata M
Nippon Roche K.K., PCR Dept.
Kansenshogaku Zasshi. 1996 Jul;70(7):710-6. doi: 10.11150/kansenshogakuzasshi1970.70.710.
A rapid and sensitive PCR-based microwell plate assay (PCR-MWP) system to detect the 16 S ribosome RNA gene of Helicobacter pylori was developed. Analytical sensitivity, evaluated with purified recombinant plasmid DNA and genomic DNA of H. pylori, was one copy of DNA per PCR. Specificity was validated with a panel of DNA from 75 kinds of microorganisms including Helicobacter showed weak positives, when 1 pg of DNA was input. Other microorganisms gave negative signals even when 100 pg of DNA was used for PCR. When compared with a Nested-PCR system to detect the urease A-subunit gene performed by a commercial reference laboratory, the results obtained (sensitivity 93.3% and specificity 73.3%) was almost equivalent. The PCR-MWP was rapid and easy for the detection of H. pylori DNA in gastric juice specimen.
开发了一种基于聚合酶链反应(PCR)的快速灵敏微孔板检测系统(PCR-MWP),用于检测幽门螺杆菌的16S核糖体RNA基因。用纯化的重组质粒DNA和幽门螺杆菌基因组DNA评估的分析灵敏度为每个PCR反应1个DNA拷贝。用包括幽门螺杆菌在内的75种微生物的一组DNA验证了特异性,当输入1 pg DNA时,幽门螺杆菌显示弱阳性。即使使用100 pg DNA进行PCR,其他微生物也给出阴性信号。与商业参考实验室进行的用于检测脲酶A亚基基因的巢式PCR系统相比,获得的结果(灵敏度93.3%,特异性73.3%)几乎相当。PCR-MWP用于检测胃液标本中幽门螺杆菌DNA快速且简便。