Multiplex PCR for the simultaneous detection of Chlamydia pneumoniae, Mycoplasma pneumoniae and Legionella pneumophila in community-acquired pneumonia

Naoyuki Miyashita, Atsushi Saito, Shigeru Kohno, Keizo Yamaguchi, Akira Watanabe, Hiroshi Oda, Yukumasa Kazuyama, Toshiharu Matsushima

    Research output: Contribution to journalArticlepeer-review

    30 Citations (Scopus)

    Abstract

    A multiplex polymerase chain reaction (PCR) was developed for the simultaneous detection of Chlamydia pneumoniae, Mycoplasma pneumoniae and Legionella pneumophila. Oligonucleotide primers for the amplification of the DNA of these three organisms were optimized for use in combination in the same reaction. PCR products were detected by the Micro-Chip Electrophoresis Analysis System. Clinical samples were obtained from 208 community-acquired pneumonia (CAP) patients who were participants in a multicenter CAP surveillance study performed at seven medical schools and their affiliate hospitals in Japan. No significant differences in the sensitivity of each primer set were observed when tested in both the multiplex and monoplex PCR assays. Our multiplex PCR was able to reliably detect 10 copies/100 μl of each of the three pathogen DNAs. Of the panel of 208 samples, 14 of 15 C. pneumoniae, 10 of 10M. pneumoniae, eight of eight L. pneumophila and 165 of 176 negative samples were correctly identified. Eleven cases who were the multiplex PCR positive and conventional method negative were observed. The PCR findings were of possible significance in at least four of these patients. Our multiplex PCR assay could potentially be used as a diagnostic and epidemiological tool. Further prospective studies are needed to establish its clinical usefulness.

    Original languageEnglish
    Pages (from-to)542-550
    Number of pages9
    JournalRespiratory Medicine
    Volume98
    Issue number6
    DOIs
    Publication statusPublished - 2004 Jun

    Keywords

    • Chlamydia pneumoniae
    • Community-acquired pneumonia
    • Legionella pneumophila
    • Multiplex PCR
    • Mycoplasma pneumoniae

    ASJC Scopus subject areas

    • Pulmonary and Respiratory Medicine

    Fingerprint Dive into the research topics of 'Multiplex PCR for the simultaneous detection of Chlamydia pneumoniae, Mycoplasma pneumoniae and Legionella pneumophila in community-acquired pneumonia'. Together they form a unique fingerprint.

    Cite this