铜绿假单胞菌的基因诊断

Study on gene diagnosis for Pseudomonas aeruginosa

目的 早期诊断铜绿假单胞菌感染。方法 采用聚合酶链反应 (PCR)检测铜绿假单胞菌OprI基因片段 ,用HaeⅢ和PvuⅡ酶切鉴定 ,测序分析。结果 培养阳性的 96份标本均扩增到 2 4 9bp片段 ,培养为其他菌及阴性的 12 7份均未发现预期扩增片段 ,经HaeⅢ和PvuⅡ酶切分别得到 4 9bp和 112bp小片段 ,测序分析其序列与基因库序列比较同源性达 10 0 %。 结论 检测铜绿假单胞菌OprI基因片段 ,可作为铜绿假单胞菌感染的早期诊断。

Objective To diagnose the infection of Pseudomonas aeruginosa early. Methods Polymerase chain reaction (PCR) was used to amplify the fragment of Pseudomonas aeruginosa OprI gene. The fragment was determined by HaeⅢ and PvuⅡ digestion, and sequencing analyses. Results It showed that 96 of 223 specimens were cultured to be positive with Pseudomonas aeruginosa, 96 of which had expectant streaks. Otherwise the other specimens had no positive streaks. The procedure needed only 4 hours. The PCR products were determined by ribonuclease HaeⅢ and PvuⅡ , and resulted in two small fragments with 49bp and 112bp separately. By automatic sequencing analysis, the coincidence rate with the gene bank was 100%. Conclusions The results indicates that the OprI gene detection by PCR is a specific, sensitive and quick technique for the diagnosis of Pseudomonas aeruginosa infection.