title: Simultaneous Detection of Nine Key Bacterial Respiratory Pathogens Using Luminex xTAG®Technology.
abstract: Early diagnosis and treatment are crucial to the outcome of lower respiratory tract infections (LRTIs). In this study, we developed an assay combining multiplex PCR and Luminex technology (MPLT) for the detection of nine important respiratory bacterial pathogens, which frequently cause LRTIs. These wereStreptococcus pneumoniae,Moraxella catarrhalis,Staphylococcus aureus,Streptococcus pyogenes,Haemophilus influenzae,Mycoplasma pneumoniae,Legionellaspp.,Pseudomonas aeruginosa, andKlebsiella pneumoniae. Through the hybridization reaction between two new synthesized multiplex PCR products and MagPlex-TAG Microspheres, we demonstrate that the detection limits for these nine pathogens were as low as 10²-10³ CFU/mL. Furthermore, 86 clinical bronchoalveolar lavage fluid specimens were used to evaluate this method. Compared with the results of nine simplex real-time PCR reactions targeting these nine pathogens, this MPLT assay demonstrated a high diagnostic accuracy forStreptococcus pneumoniae(sensitivity, 87.5% and specificity, 100%). Furthermore, sensitivity and specificity for the other eight pathogens all attained 100% diagnostic accuracy. In addition, the consistency between MPLT and the nine real-time PCR reactions exceeded 98.8%. In conclusion, MPLT is a high-throughput, labor-saving and reliable method with high sensitivity and specificity for identifying nine respiratory pathogens responsible for LRTIs. Indeed, this assay may be a promising supplement to conventional methods used to diagnose LRTIs.
authors.affiliation: State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China. jlx67@126.com.|Department of Respiratory Medicine, Shengjing Hospital of China Medical University, Shenyang 110004, China. jlx67@126.com.|Department of Respiratory Medicine, Zhejiang Provincial People's Hospital, Hangzhou 310014, China. jlx67@126.com.
authors.forename: Luxi
authors.initials: L
authors.lastname: Jiang
authors.affiliation: State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China. renhongyu@icdc.cn.
authors.forename: Hongyu
authors.initials: H
authors.lastname: Ren
authors.affiliation: State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China. zhj_0901@163.com.
authors.forename: Haijian
authors.initials: H
authors.lastname: Zhou
authors.affiliation: State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China. qintian@icdc.cn.
authors.forename: Tian
authors.initials: T
authors.lastname: Qin
authors.affiliation: Department of Respiratory Medicine, Shengjing Hospital of China Medical University, Shenyang 110004, China. chenyusy@hotmail.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Chen
mesh_terms: D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology*; D002681:China; D006193:Haemophilus influenzae / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D007711:Klebsiella pneumoniae / Q000235:genetics / Q000302:isolation & purification; D007875:Legionella / Q000235:genetics / Q000302:isolation & purification; D001936:Moraxella catarrhalis / Q000235:genetics / Q000302:isolation & purification; D060885:Multiplex Polymerase Chain Reaction; D009177:Mycoplasma pneumoniae / Q000235:genetics / Q000302:isolation & purification; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D012141:Respiratory Tract Infections / Q000175:diagnosis* / Q000382:microbiology*; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification; D013296:Streptococcus pneumoniae / Q000235:genetics / Q000302:isolation & purification; D013297:Streptococcus pyogenes / Q000235:genetics / Q000302:isolation & purification
keywords: Luminex; detection method; lower respiratory tract infections; multiplex PCR; respiratory pathogens