铜绿假单胞菌oprI基因的反向斑点杂交检测法

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100ng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。

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

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 bacteria, 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.