随机扩增多态DNA指纹技术的研究及在检测医院感染中的应用

A randomized Study on amplified Polymorphic DNA Assay and its Application in Hospital Infection

目的 探讨医院感染常见病原菌的基因鉴定及分型方法 ,了解其分子流行病学情况。方法 优化了随机扩增多态 DNA指纹技术 (RAPD)的实验条件 ;利用 RAPD技术成功地检测了儿科病房呼吸道合胞病毒 (RSV)、脑外科病房铜绿假单胞菌 (PA)及血液病房白色念珠菌 (CA)的医院感染。同时进行了 RSV分离、PA菌和 CA菌培养。结果 构建了金黄色葡萄球菌 (SA )、志贺菌 (SB)、大肠埃希菌 (EC)、肠炎沙门菌 (Salm )、铜绿假单胞菌(PA)、白色念珠菌 (CA) 3、7、11型腺病毒 (Ad3、Ad7、Ad11)、呼吸道合胞病毒 (RSV)的图谱 ;用 RAPD法从 76份疑似 RSV医院感染患儿的鼻咽分泌物中分离出 4 9株 RSV,共有 4种基因型 ,R1 型占 4 4株 ,在同一疗区同一时期的 R1 3 8株。RAPD法与分离法比较阳性符合率 10 0 % ,阳性检出率高 ,P<0 .0 1;从 2名脑外科术后感染患者脑脊液及相关标本中检出 PA菌 ;从肿瘤患者化疗后感染血液中检出 CA菌。结论

OBJECTIVE To study hospital infection bacteria gene types and to know about their molecular epidemiology. METHODS Using RAPD technology to monitor Pseudomonas aeruginosa (PA) in neurosergical ward, Candidiasis albicans (CA) in cancer patients treated by chemotherapy and respiratory syncytial virus (RSV) in pediatric respiratory disease ward. RESULTS We found four PA strains, 21 CA straims; 49 RSV strains and there were four gene types, R 1 was the strain which caused hospital infection. RAPD compared isolating method gave the corcesponding rate 100% and the positive rate was higer than non isolating (P<0.01). CONCLUSION RAPD may be a good method to monitor nosocomial infection. It is specific and sensitive.