摘要
在HP高度保守的尿素酶A基因内合成两对引物,建立了巢式聚合酶链反应(N-PCR)检测HPDNA的方法,实验证明有很高的特异性和敏感性,与其它4株肠道菌无交叉反应,最小检出量为1fgHPDNA。对N-PCR反应体系中有关实验条件如Mg￣2+浓度、引物浓度、复性温度等进行优化选择,并对酚/氯仿提取法和裂解粗提法两种标本处理方法进行了比较。用建立的N-PCR检测30例有上消化道症状患者的唾液及8例胃组织标本中HPDNA,结果阳性率分别为56.67%和62.50%,5例胃组织标本阳性患者中4例唾液标本亦阳性。实验表明N-PCR是一种敏感特异的检测方法,可用于唾液等标本中HP的检测。
A nested polymerase chain reaction(N-PCR),involving two pairs of primers deduced from the highly conserved urease A gene of Helicobacter pylori(HP),was develped for detecting HP DNA.The specificity and sensitivity of N-PCR was proved.There was no cross-reaction with other 4 enteric strains.The lowest detection level of N-PCR was lfg HP DNA.The experiment conditions were optimized including the Mg^(2+)concentration,primer concentration,annealing temperature and the method of samples handling.30 saliva samples and 8 gastric biopsy materials from patients with various gastric disease were examined by N-PCR and the positive rates of HP DNA were 56.67%and 62.50%,respectively.4 of 5 patients who tested positive by N-PCR of gastric biopsy materials were positive when a salive sample was analyzed.The results shows the N-PCR was a sentitive and specific method and could be used to detect HP in saliva sample.
出处
《空军总医院学报》
1996年第3期135-137,共3页
Journal of General Hospital of Air Force,PLA
