多重荧光PCR应用于下呼吸道感染病原菌检测的研究

Detection of Pathogenic Bacteria in Lower Respiratory Tract Infection by Multiplex Fluorescent PCR

目的应用多重荧光PCR技术检测下呼吸道感染常见病原菌,为下呼吸道感染提供一种高效的诊断工具。方法建立多重荧光PCR法检测下呼吸道感染常见细菌中的大肠埃希菌、肺炎克雷伯菌、铜绿假单胞菌和肺炎链球菌,应用该法检测了112例临床痰标本,并与细菌培养结果进行比对。结果在多重PCR反应体系中,大肠埃希菌、肺炎克雷伯菌、铜绿假单胞菌的灵敏度为5×102 copies/ml,肺炎链球菌的灵敏度为2.5×102 copies/ml;多重荧光PCR检测112例临床痰标本,大肠埃希菌阳性率为9.8%,肺炎克雷伯菌阳性率为12.5%,铜绿假单胞菌阳性率为17.0%,肺炎链球菌阳性率为0.9%。结论多重荧光PCR检测方法具有灵敏度高、特异性强、通量高和速度快等优点,为下呼吸道感染诊断提供了新的检测手段。

Objective To establish a highly efficient technique of diagnosis to detect common Pathogenic bacteria leading to lower respiratory tract infections using multiple fluorescence PCR technique. Methods Using multiple fluorescent PCR to detect common bacteria leading to the lower respiratory tract infection such as e. coli, klebsiella pneumoniae, pseudomonas aeruginosa and s. pneumonia in the 112 samples of clinical sputum specimens, and compare with the results of bacterial culture. Results The sensitivity of the test by multiple PCR method for e. coli, klebsiella pneumonia and pseudomonas aeruginosa was 5 × 102 copies/ml, and for streptococcus pneumonia was 2.5 × 102 copies/ml. In 112 sputum specimens collected from clinical respiratory tract infection, the positive rate of e. coli klebsiella pneumonia, pseudomonas aeruginosa and streptococcus pneumonia were 9.8%, 12.5%, 17.0% and 0.9% respectively. Conclusion Multiplex fluorescent PCR detection method has the advantages of high sensitivity, high specificity, high throughput and high speed. It provides a new detection method for the diagnosis of lower respiratory tract infections.