Kai M, Kamiya S, Yabe H, Takakura I, Shiozawa K, Ozawa A
Department of Infectious Diseases, School of Medicine, Tokai University, Kanagawa, Japan.
J Med Microbiol. 1993 Mar;38(3):166-70. doi: 10.1099/00222615-38-3-166.
A DNA amplification method was used to detect Mycoplasma pneumoniae in clinical samples. M. pneumoniae 16S ribosomal RNA gene sequences were selected as the amplification target region. The polymerase chain reaction (PCR) with purified DNA fragments as templates yielded an expected 88-bp fragment from M. pneumoniae but not from other Mycoplasma spp. nor from any of the other bacteria assayed. With this method, the 88-bp product specific for M. pneumoniae could be obtained from a minimum of 0.05 pg of M. pneumoniae DNA. Subsequently this PCR technique was used for the detection of M. pneumoniae in throat-swab samples. Twenty-two of 30 culture-positive clinical samples gave positive results in the PCR test. Thirty-two culture-negative clinical samples and 33 samples from healthy volunteers, of which only one was culture-positive, gave negative results in the same PCR test. This PCR method is useful for the direct detection of M. pneumoniae in clinical samples.
采用DNA扩增方法检测临床样本中的肺炎支原体。选择肺炎支原体16S核糖体RNA基因序列作为扩增靶区域。以纯化的DNA片段为模板进行聚合酶链反应(PCR),可从肺炎支原体中产生预期的88bp片段,而其他支原体属以及所检测的任何其他细菌均未产生该片段。用此方法,最少0.05pg的肺炎支原体DNA即可获得肺炎支原体特异性的88bp产物。随后,该PCR技术用于检测咽拭子样本中的肺炎支原体。30份培养阳性的临床样本中有22份在PCR检测中呈阳性结果。32份培养阴性的临床样本以及33份来自健康志愿者的样本(其中只有1份培养阳性)在同一PCR检测中均呈阴性结果。该PCR方法可用于直接检测临床样本中的肺炎支原体。