聚合酶链反应快速检测胃组织和唾液中的幽门螺杆菌

Rapid detection of helicobacter pylori in gastric biopsy and saliva samples by polymerase chain reaction

针对幽门螺杆菌（HP）尿素酶A基因设计一对引物进行聚合酶链反应，检测1株HP标准株和7株临床分离株均阳性，而4株其它肠道菌均阴性，特异性100％。10倍系列稀释试验表明敏感性达到100pgDNA水平。从35例胃镜检查者取幽门旁组织块进行快速和常规尿素酶试验，细菌培养及PCR检测，15例HP阳性者PCR检测也为阳性，其中7例阳性者有3例唾液PCR检测为阳性，表明HP确存在于口腔中。本研究采用直接热裂解法处理临床标本，取其粗提物行PGR免除复杂的酚-氯仿抽提步骤，该法简便快速，且损失小，成功率高，在临床实验诊断中有推广价值。

By using primers based on the sequence of urease A gene of HP, a protocol was established to detect this bacterium in gastric biopsy and saliva samples by PCR. This assay could detect all 8 HP strains, but failed to detect other 4 enteric strains, showing it to be 100% specific. Tenfold serial dilution experiments revealed the detection of as little as 100pg DNA by this assay. HP was detected by PCR in 15 gastric biopsies from patients positive for it, and only 3 of 7 patients who tested positive by PCR of gastric biopsy were positive when a saliva sample was analyzed. In this assay, crude extraction by single-step heat lysis of clinical samples was developed, instead of multi-step phenol chloroform extraction. The results showed it to be simple, rapid and successful, thus its value as a laboratory diagnostic test was comfirmed.