Thong K L, Lai M Y, Teh C S J, Chua K H
Laboratory of Biomedical Science and Molecular Microbiology, Institute of Graduate Studies, University of Malaya, 50603, Kuala Lumpur, Malaysia.
Trop Biomed. 2011 Apr;28(1):21-31.
A PCR-based assay that can simultaneously detect and differentiate five different types of nosocomial bacterial pathogens was developed. Six pairs of selected primers targeting femA (132 bp) and mecA (310 bp) of methicillin-resistant Staphylococcus aureus, gltA (722 bp) of Acinetobacter baumannii, phoA (903 bp) of Escherichia coli, mdh (364 bp) of Klebsiella pneumoniae and oprL (504 bp) of Pseudomonas aeruginosa were used in this study. The conditions were optimized for the multiplex PCR to ensure specific amplification of the selected targets. Sensitivity and specificity tests were also carried out using a blind test approach on 50 bacterial cultures and resulted in 100% for both positive and negative predictive values.
开发了一种基于聚合酶链反应（PCR）的检测方法，该方法可同时检测和区分五种不同类型的医院内细菌病原体。本研究使用了六对选定的引物，分别靶向耐甲氧西林金黄色葡萄球菌的femA（132 bp）和mecA（310 bp）、鲍曼不动杆菌的gltA（722 bp）、大肠杆菌的phoA（903 bp）、肺炎克雷伯菌的mdh（364 bp）以及铜绿假单胞菌的oprL（504 bp）。对多重PCR的条件进行了优化，以确保选定靶标的特异性扩增。还采用盲法对50份细菌培养物进行了敏感性和特异性测试，阳性和阴性预测值均为100%。
