Development of a PCR-based method for diagnosing mycoplasma pneumoniae infections

Current methodologies for detecting and identifying Mycoplasma pneumoniae are time consuming and difficult. PCR was shown to be most valuable in detection and identification of such cases. Nevertheless, false-negative PCR results are rather common due to inhibitors of the PCR reaction in the clinical specimen, while false-positive results may occur due to contamination of the reagents with target DNA. To overcome these problems, we designed two nested pairs of oligonucleotide primers which anneal to DNA sequences coding for conserved regions of the 16 SrRNA. The nested PCR reacted with all of the strains of Mycoplasma pneumoniae, and gave no amplification product with any of the closely related mycoplasmas tested, showing its high specificity. Thus the PCR-based method for the detection of Mycoplasma pneumoniae was shown to be useful in clinical diagnosis.