非同位素标记脆弱类杆菌DNA探针的研究  

A study of non radioactive DNA probe for detection of Bacteroides fragilis

作　　者：汪冰[1] 黄安华[1] 冯萍[1] 范昕建[1] 朱淑媛[1] 曾维华[1] 曹钟梁[1] 

机构地区：[1]成都华西医科大学

出　　处：《中华传染病杂志》1997年第1期9-11,共3页Chinese Journal of Infectious Diseases

摘　　要：为探索应用分子生物学技术检测厌氧菌的方法，选择脆弱类杆菌抽提其染色体ＤＮＡ，用生物素和地高辛配基标记制备ＤＮＡ探针，检测厌氧和需氧菌９４株，临床标本４５份，并和３２Ｐ标记探针及培养生化鉴定比较。该探针可检出临床上常见的类杆菌属细菌，与其它细菌无交叉反应。结果：地高辛配基标记探针的灵敏度（１０ｐｇＤＮＡ和１０３ＣＦＵ／ｍｌ）接近３２Ｐ标记探针的灵敏度（１ｐｇＤＮＡ和１０３ＣＦＵ／ｍｌ）。地高辛配基和生物素化的敏感性为８９．５％和５３．６％，特异性分别为９２．３％和８４．６％。结果表明脆弱类杆菌染色体ＤＮＡ探针可检测临床上常见的类杆菌属厌氧菌，敏感性和特异性较高。This study was designed to explore a rapid, specific sensitive and simple molecular biological method which was applied for the detection of anagrobes. In this study we chose the commom anaerobes in clinic Bacteroides fragilis(B.f)to extract the total DNA and prepare the DNA probes labeled with non radioactive biotin and digoxigenin. 94 clinical isolates from anaarobes and aerobes as well as 45 clinical specimens were tested with these probes, compared with 32p labeled probes, anaerobic culture and biochemical reactions. The results showed that the common Bacteroides in clinic might be detected with these probes. No cross reactions with other anaerobes and aerobes were found. The sensitivity of digoxigeninated probes was 10 pg of DNA and 10 3 CFU/ml of Bacteroides, Comparable to 32p labeld probes (1 pg of DNA and 10 3 CFU/ml)and higher than biotinated probes (100pg DNA and 10 4 CFU/ml). When compared with culture and biochemical identification of bacteroides, the sensitivity of digoxigeninated and biotinated probes were 89.47% and 53.63%, the specificity were 92.31% and 84.61% respectively. These data revealed that the chromosomal DNA probes of Bacteroides fragilis could detect the common clinical Bacteroides with high sensitivity and specificity.

关 键 词：非同位素标记 脆弱类杆菌 DNA探针 

分 类 号：R446.5[医药卫生—诊断学]