[应用反向斑点杂交技术检测铜绿假单胞菌oprI基因]
[Detection of oprI gene of Pseudomonas aeruginosa by reverse dot-blot hybridization].
作者信息
Fan Hui-zhen, Huang Wen-jie, Liang Kun, Fang Yi, Ma Li-ren, Liu Yao-qing
机构信息
Department of Respiratory Diseases, Guangzhou General Hospital of Guangzhou Command, Guangzhou 510010, China.
出版信息
Di Yi Jun Yi Da Xue Xue Bao. 2004 Mar;24(3):303-5.
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.
目的
建立一种在感染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oprI基因序列设计特异性引物，通过PCR合成特异性探针。对细菌DNA进行光敏生物素标记后，采用反向斑点杂交法检测铜绿假单胞菌。
结果
合成的探针对铜绿假单胞菌具有高度特异性，与其他细菌、病毒或真菌无交叉反应。该方法能够检测100 ng细菌DNA。
结论
反向斑点杂交法在感染早期检测铜绿假单胞菌方面具有快速、特异的优点。
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