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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００１ｐｇ．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．
Objective To establish a polymerase chain reaction (PCR) for the amplification of HpDNA from paraffin-embedded gastric mucosa specimens. Methods A pair of Hp-specific oligonucleotides was used as a primer to amplify the Hp16SrRNA gene by double-amplification PCR and compared with routine detection methods. Results The minimum amount of HpDNA detected by double amplification PCR was 0. 01 pg. Using this method to detect 20 cases of paraffin-embedded gastric mucosa in patients with duodenal ulcer, and compared with urease test, bacterial culture, Warthin_Starry silver staining and PCR detection of fresh gastric mucosa specimens, 18 patients in the above five Two cases of urease test, silver staining and PCR in fresh gastric mucosa specimens were positive in patients, using double amplification PCR detection of paraffin-embedded specimens found no HpDNA. PCR detection of paraffin-embedded specimens and fresh gastric mucosa specimens have a significant correlation. Conclusion Double-amplification PCR provides an advantageous tool for the retrospective study of Hp infection at the molecular level.