title: Association of targeted multiplex PCR with resequencing microarray for the detection of multiple respiratory pathogens.
abstract: A large number of viral and bacterial organisms are responsible for community-acquired pneumonia (CAP) which contributes to substantial burden on health management. A new resequencing microarray (RPM-IVDC1) associated with targeted multiplex PCR was recently developed and validated for multiple respiratory viruses detection and discrimination. In this study, we evaluated the capability of RPM-IVDC1 for simultaneous identification of multiple viral and bacterial organisms. The nasopharyngeal aspirates (NPAs) of 110 consecutive CAP patients, aged from 1 month to 96 years old, were collected from five distinct general hospitals in Beijing during 1-year period. The samples were subjected to the RPM-IVDC1 established protocol as compared to a real-time PCR (qRT-PCR), which was used as standard. The results of virus detection were consistent with those previously described. A total of 37 of Streptococcus pneumoniae, 14 of Haemophilus influenzae, 10 of Mycoplasma pneumoniae, two of Klebsiella pneumoniae and one of Moraxella catarrhalis were detected by RPM-IVDC1. The sensitivities and specificities were compared with those of qRT-PCR for S. pneumoniae (100, 100%, respectively), H. influenzae (92.3, 97.9%, respectively), M. pneumoniae (69.2, 99.0%, respectively), K. pneumoniae (100, 100%, respectively), and M. catarrhalis (100, 100%, respectively). Additional 22 of Streptococcus spp., 24 of Haemophilus spp. and 16 of Neisseria spp. were identified. In addition, methicillin-resistant and carbapenemases allele were also found in nine of Staphylococcus spp. and one of K. pneumoniae, respectively. These results demonstrated the capability of RPM-IVDC1 for simultaneous detection of broad-spectrum respiratory pathogens in complex backgrounds and the advantage of accessing to the actual sequences, showing great potential use of epidemic outbreak investigation. The detection results should be carefully interpreted when introducing this technique in the clinical diagnostics.
authors.affiliation: Key Laboratory of Medical Virology, Ministry of Health, Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention Beijing, China ; Futian District Center for Disease Control and Prevention Shenzhen, China.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Shen
authors.affiliation: State Key Laboratory for Infectious Disease Prevention and Control, Chinese Center for Disease Control and Prevention, National Institute for Communicable Disease Control and Prevention Beijing, China.
authors.forename: Bingqing
authors.initials: B
authors.lastname: Zhu
authors.affiliation: Third Hospital of Beijing Armed Police Corps Laboratory Beijing, China.
authors.forename: Shulian
authors.initials: S
authors.lastname: Wang
authors.affiliation: Futian District Center for Disease Control and Prevention Shenzhen, China.
authors.forename: Haolian
authors.initials: H
authors.lastname: Mo
authors.affiliation: Key Laboratory of Medical Virology, Ministry of Health, Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention Beijing, China.
authors.forename: Ji
authors.initials: J
authors.lastname: Wang
authors.affiliation: Key Laboratory of Medical Virology, Ministry of Health, Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention Beijing, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Medical Virology, Ministry of Health, Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention Beijing, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Futian District Center for Disease Control and Prevention Shenzhen, China.
authors.forename: Huashu
authors.initials: H
authors.lastname: Zeng
authors.affiliation: Key Laboratory of Medical Virology, Ministry of Health, Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention Beijing, China.
authors.forename: Li
authors.initials: L
authors.lastname: Guan
authors.affiliation: Beijing Center for Disease Control and Prevention, Institute for Infectious Disease and Endemic Disease Control Beijing, China.
authors.forename: Weixian
authors.initials: W
authors.lastname: Shi
authors.affiliation: Futian District Center for Disease Control and Prevention Shenzhen, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Medical Virology, Ministry of Health, Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention Beijing, China.
authors.forename: Xuejun
authors.initials: X
authors.lastname: Ma
keywords: RPM-IVDC1; community-acquired pneumonia; detection; multiple respiratory pathogens; resequencing microarray