Simultaneous Detection of Key Bacterial Pathogens Related to Pneumonia and Meningitis Using Multiplex PCR Coupled With Mass Spectrometry

Zhang, Chi; Xiu, Leshan; Xiao, Yan; Xie, Zhengde; Ren, Lili; Peng, Junping

Based on the sensitivity test, the limit of detection (LOD) of the BP-MS method is 10 gene copies/reaction. In this study, 31 samples positive for at least one of the target pathogens using the BP-MS method were not confirmed positive by either realtime PCR or nested PCR. We speculate that the different designs of the BP-MS method and confirmatory tests accounted for such discrepancies. Apart from methodological factors, the lower LOD with the BP-MS method may be attributable to the five additional cycles of primary multiplex PCR and dissimilar target selection (H. influenzae and S. aureus, Table S2 ). For target pathogens with low rates of concordance, such as K. pneumoniae, this disagreement may be attributable to differences in sensitivities of the BP-MS method and confirmatory tests.