Woods C R, Versalovic J, Koeuth T, Lupski J R
Department of Pediatrics, Baylor College of Medicine, Houston, Texas 77030.
J Clin Microbiol. 1993 Jul;31(7):1927-31. doi: 10.1128/jcm.31.7.1927-1931.1993.
Repetitive element sequence-based polymerase chain reaction (rep-PCR) enables the generation of DNA fingerprints which discriminate bacterial species and strains. We describe the application of whole-cell methods which allow specimens from broth cultures or colonies from agar plates to be utilized directly in the PCR mixture. The rep-PCR-generated DNA fingerprints obtained with whole-cell samples match results obtained with genomic DNA templates. Examples with different gram-negative bacteria (e.g., Citrobacter diversus, Escherichia coli, and Pseudomonas aeruginosa) and gram-positive bacteria (e.g., Staphylococcus aureus and Streptococcus pneumoniae) are demonstrated. Rapid specimen preparation methods enable rep-PCR-based fingerprinting to be completed in several hours and, therefore, allows the timely analysis of epidemiological relationships.
基于重复元件序列的聚合酶链反应(rep-PCR)能够生成区分细菌种类和菌株的DNA指纹图谱。我们描述了全细胞方法的应用,该方法允许直接将肉汤培养物中的标本或琼脂平板上的菌落用于PCR混合物中。用全细胞样本获得的rep-PCR生成的DNA指纹图谱与用基因组DNA模板获得的结果相匹配。文中展示了不同革兰氏阴性菌(如奇异柠檬酸杆菌、大肠杆菌和铜绿假单胞菌)和革兰氏阳性菌(如金黄色葡萄球菌和肺炎链球菌)的实例。快速的标本制备方法使基于rep-PCR的指纹图谱分析能在数小时内完成,因此能够及时分析流行病学关系。