title: Simultaneous Detection of Five Pathogens from Cerebrospinal Fluid Specimens Using Luminex Technology.
abstract: Early diagnosis and treatment are crucial for the outcome of central nervous system (CNS) infections. In this study, we developed a multiplex PCR-Luminex assay for the simultaneous detection of five major pathogens, including Mycobacterium tuberculosis, Cryptococcus neoformans, Streptococcus pneumoniae, and herpes simplex virus types 1 and 2, which frequently cause CNS infections. Through the hybridization reaction between multiplex PCR-amplified targets and oligonucleotide "anti-TAG" sequences, we found that the PCR-Luminex assay could detect as low as 10¹-10² copies of synthetic pathogen DNAs. Furthermore, 163 cerebrospinal fluid (CSF) specimens from patients with suspected CNS infections were used to evaluate the efficiency of this multiplex PCR-Luminex method. Compared with Ziehl-Neelsen stain, this assay showed a high diagnostic accuracy for tuberculosis meningitis (sensitivity, 90.7% and specificity, 99.1%). For cryptococcal meningitis, the sensitivity and specificity were 92% and 97.1%, respectively, compared with the May Grunwald Giemsa (MGG) stain. For herpes simplex virus types 1 and 2 encephalitis, the sensitivities were 80.8% and 100%, and the specificities were 94.2% and 99%, respectively, compared with Enzyme Linked Immunosorbent Assay (ELISA) assays. Taken together, this multiplex PCR-Luminex assay showed potential efficiency for the simultaneous detection of five pathogens and may be a promising supplement to conventional methods for diagnosing CNS infections.
authors: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. zhoelfu@163.com.|Department of Neurology, Third Hospital of People's Liberation Army, Baoji 721004, China. zhoelfu@163.com.
authors: Linfu
authors: L
authors: Zhou
authors: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. wurui1124@126.com.
authors: Rui
authors: R
authors: Wu
authors: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. shixd999@163.com.
authors: Xiaodan
authors: X
authors: Shi
authors: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. fengdy@fmmu.edu.cn.
authors: Dongyun
authors: D
authors: Feng
authors: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. fgd2000@fmmu.edu.cn.
authors: Guodong
authors: G
authors: Feng
authors: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. yiningyang80@163.com.
authors: Yining
authors: Y
authors: Yang
authors: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. wwdd126@126.com.
authors: Wen
authors: W
authors: Dai
authors: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. bianting-44@163.com.
authors: Ting
authors: T
authors: Bian
authors: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. miracle6tt@163.com.
authors: Tingting
authors: T
authors: Liu
authors: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. skyblue20090802@163.com.
authors: Ying
authors: Y
authors: He
authors: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. biomidas@163.com.
authors: Ming
authors: M
authors: Shi
authors: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. zhaogang@fmmu.edu.cn.
authors: Gang
authors: G
authors: Zhao
mesh_terms: D002494:Central Nervous System Infections / Q000175:diagnosis; D002555:Cerebrospinal Fluid / Q000382:microbiology*; D003455:Cryptococcus neoformans / Q000302:isolation & purification*; D018259:Herpesvirus 1, Human / Q000302:isolation & purification*; D018258:Herpesvirus 2, Human / Q000302:isolation & purification*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D009169:Mycobacterium tuberculosis / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013296:Streptococcus pneumoniae / Q000302:isolation & purification*
keywords: Luminex technology; central nervous system infections; cerebrospinal fluid; multiplex PCR