胃粘膜石蜡切片PCR法检测幽门螺杆菌

目的建立从石蜡包埋的胃粘膜标本中扩增ＨｐＤＮＡ的聚合酶链反应（ＰＣＲ）．方法以一对Ｈｐ特异的寡核苷酸为引物，采用双扩增ＰＣＲ扩增Ｈｐ１６ＳｒＲＮＡ基因，并与常规检测方法进行了比较．结果双扩增ＰＣＲ检出的最小ＨｐＤＮＡ量为００１ｐｇ．用该方法检测十二指肠溃疡患者２０例石蜡包埋的胃粘膜标本，并与尿素酶试验、细菌培养、Ｗａｒｔｈｉｎ＿Ｓｔａｒｒｙ银染色及ＰＣＲ对新鲜胃粘膜标本的检测作比较，结果１８例患者上述五项检测结果一致，２例尿毒酶试验、银染色及ＰＣＲ对新鲜胃粘膜标本的检测呈阳性的患者，采用双扩增ＰＣＲ对石蜡包埋标本的检测未发现ＨｐＤＮＡ．ＰＣＲ对石蜡包埋标本及新鲜胃粘膜标本的检测有显著相关性．结论双扩增ＰＣＲ为分子水平上Ｈｐ感染的回顾性研究提供一有利工具．

PCR detection of Helicobacter pylori in paraffin embedded gastric mucosa

AIM To establish the PCR method to amplify H.pylori chromosomal DNA from paraffin embedded gastric mucosa. METHODS A specific oligonucleotide primer pair was used to amplify the 16S rRNA gene of H.pylori by double PCR. The results were compared with that of routine methods in 20 gastric mucosal specimens. RESULTS Double PCR amplification can detect a minimun of 0 01pg H.pylori DNA. By comparing double PCR in paraffin embedded specimens of 20 duodenal ulcer patients with urease test, culture, Warthin_Starry stain and PCR in fresh specimens, we demonstrated that 18 patients had the same results by these five tests. Only 2 patients infected with H.pylori were positive by urease test, Warthin_Starry stain and PCR with fresh specimens, but negative by double PCR with paraffin embedded specimens. Significant association was found between the results detected by PCR with paraffin embedded specimens and that with fresh specimens. CONCLUSION Double PCR provides a useful method for retrospective research of H.pylori infection on the molecular level.