Development of a multiplex real-time PCR assay for the simultaneous detection of four bacterial pathogens causing pneumonia

1 answer:

4 0

To simultaneously identify the four bacterial pathogens that cause pneumonia, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, and Moraxella catarrhalis, a multiplex real-time polymerase chain reaction (mRT-PCR) technique was developed.

Limit of detection (LOD), specificity, and reproducibility were used to assess the analytical performance of mRT-PCR against target pathogens. For the "respiratory bacteria four" (RB4) mRT-PCR assay, 210 clinical samples from pneumonia patients were processed using automatic nucleic acid extraction equipment.

The outcomes were then directly compared to references from bacterial culture and/or Sanger sequencing. With a coefficient of variance ranging from 0.29 to 1.71 and a cautious LOD of DNA equating to 5 102 copies/reaction, the RB4 mRT-PCR test identified all target pathogens from sputum samples.

Additional bacterial infections were found in reference-negative specimens, and the assay had 100% agreement with reference-positive samples.

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