Okazaki N, Yamai S, Sasaki Y, Sasaki T
Kanagawa Prefectural Public Health Laboratories, Department of Bacteriology and Pathology.
Kansenshogaku Zasshi. 1998 Jul;72(7):742-6. doi: 10.11150/kansenshogakuzasshi1970.72.742.
Two-step polymerase chain reaction (PCR) with primers designated against 16S rRNA gene of Mycoplasma pneumoniae for diagnosis of infection was evaluated in comparison with the conventional single-step PCR and culture methods. The two-step PCR method showed specific amplification of M. pneumoniae DNA and higher sensitivity (1.5 fg/assay) than the single-step PCR method. With the two-step PCR method, 76 of 322 throat swabs (23.6%) from patients with acute respiratory complaints gave positive results whereas 20.2% were positive in the culture method. Seven of 13 samples which were negative in the single-step PCR method but positive in either serological or the culture method showed positive results by the two-step PCR method. In addition, 5 samples which were weakly positive in the single-step PCR method showed distinctly positive results in the two-step PCR. These results indicate that the two-step PCR method is a useful tool for detection of M. pneumoniae in clinical specimens, although it requires a relatively sophisticated in technique.
为诊断感染,对针对肺炎支原体16S rRNA基因设计引物的两步聚合酶链反应(PCR)与传统单步PCR和培养方法进行了比较评估。两步PCR方法显示出肺炎支原体DNA的特异性扩增,并且比单步PCR方法具有更高的灵敏度(每次检测1.5 fg)。采用两步PCR方法,322例急性呼吸道疾病患者的咽拭子中有76例(23.6%)呈阳性结果,而培养法的阳性率为20.2%。在单步PCR方法中为阴性但血清学或培养法为阳性的13份样本中,有7份通过两步PCR方法显示出阳性结果。此外,在单步PCR方法中弱阳性的5份样本在两步PCR中显示出明显阳性结果。这些结果表明,两步PCR方法是检测临床标本中肺炎支原体的有用工具,尽管它需要相对复杂的技术。