Evaluation of the Allplex PneumoBacter Assay for the Detection of Bacterial Respiratory Infections

Respiratory tract infections, caused by bacteria and viruses, are among the most common infectious diseases worldwide. Rapid and accurate diagnostics are vital for effective treatment and infection control. While traditional methods like culture and microscopy are still used, molecular diagnostics especially multiplex Polymerase Chain Reaktion (PCR) have become increasingly important due to their ability to detect multiple pathogens at once. This study evaluated the Allplex PneumoBacter Assay, a multiplex PCR test, designed to detect bacterial respiratory pathogens, and compared its performance with the reference method, BioFire. The evaluation included clinical samples, known bacterial strains, spiked negative sputum and bronchoalveolar lavage (BAL) samples, and external quality controls. Nucleic acid extraction was performed using the STARMag 96 x 4 Universal Cartridge Kit on the Starlet IVD system, followed by PCR amplification on the CFX96 instrument using the Allplex assay. The assay showed high sensitivity (94.1%) and perfect specificity (100%) for Mycoplasma pneumoniae, Chlamydia pneumoniae, and Bordetella pertussis. The positive predictive value (PPV) was 100%, and the negative predictive value (NPV) was 95.5%. Additionally, Haemophilus influenzae and Streptococcus pneumoniae were detected in some samples that were negative using the reference method . In particular, S. pneumoniae was found in several negative sputum samples, possibly indicating early or mild infections missed by the reference method. In conclusion, the Allplex assay demonstrated strong agreement with the reference test and showed potential for broader pathogen detection. Further studies with more clinical samples are recommended to confirm its clinical value.