铜绿假单胞菌oprI基因的反向斑点杂交检测法  

Detection of oprI gene of Pseudomonas aeruginosa by reverse dot-blot hybridization

作　　者：樊慧珍[1] 黄文杰[1] 梁昆[1] 方怡[1] 马立人[2] 刘耀清[2] 

机构地区：[1]广州军区广州总医院呼吸科,广东广州510010 [2]军事医学科学院放射医学研究所,北京100850

出　　处：《第一军医大学学报》2004年第3期303-305,共3页Journal of First Military Medical University

基　　金：广东省社会发展攻关基金(2002C30405)~~

摘　　要：目的建立一种早期快速检测铜绿假单胞菌的方法。方法根据铜绿假单胞菌特异的oprI基因设计引物,聚合酶链反应合成特异探针,光敏生物素标记细菌DNA,应用反向斑点杂交法检测铜绿假单胞菌。结果所合成的探针具有高度特异性,能鉴别铜绿假单胞菌,与其他细菌、病毒、真菌间无交叉反应。该方法能检测出100 ng细菌DNA。结论反向斑点杂交法具有快速、特异的优点,对铜绿假单胞菌的早期检测具有重要意义。Objective To establish a rapid method for detecting Pseudomonas aeruginosa at the early stage of infection. Methods Specific primers were designed according to oprI gene sequence of Pseudomonas aeruginosa, and the specific probe was synthesized by PCR. After photosensitive biotin labeling of the bacterial DNA, reverse dot-blot hybridization was used to detect Pseudomonas aeruginosa. Results The probe synthesized was highly specific to Pseudomonas aeruginosa without cross reaction with other bateria, viruses or fungi. The method was capable of detecting 100 ng bacteria DNA. Conclusion Reverse dot-blot hybridization possesses the merits of speediness and specificity in the detection of Pseudomonas aeruginosa in the early stage of infection.

关 键 词：铜绿假单胞菌 oprI基因 反向斑点杂交检测法 呼吸道感染 诊断 

分 类 号：R446.5[医药卫生—诊断学]