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Int. J. Environ. Res. Public Health 2016, 13(2), 193; doi:10.3390/ijerph13020193

Simultaneous Detection of Five Pathogens from Cerebrospinal Fluid Specimens Using Luminex Technology

1
Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, China
2
Department of Neurology, Third Hospital of People’s Liberation Army, Baoji 721004, China
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed.
Academic Editor: Paul B. Tchounwou
Received: 5 December 2015 / Revised: 25 January 2016 / Accepted: 29 January 2016 / Published: 4 February 2016
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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.
Keywords: Luminex technology; multiplex PCR; central nervous system infections; cerebrospinal fluid Luminex technology; multiplex PCR; central nervous system infections; cerebrospinal fluid
This is an open access article distributed under the Creative Commons Attribution License (CC BY 4.0).

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Zhou, L.; Wu, R.; Shi, X.; Feng, D.; Feng, G.; Yang, Y.; Dai, W.; Bian, T.; Liu, T.; He, Y.; Shi, M.; Zhao, G. Simultaneous Detection of Five Pathogens from Cerebrospinal Fluid Specimens Using Luminex Technology. Int. J. Environ. Res. Public Health 2016, 13, 193.

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