Publication: Single tube multiplex real-time PCR for the rapid detection of herpesvirus infections of the central nervous system
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Issued Date
2011-04-01
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ISSN
08908508
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2-s2.0-79955452214
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Mahidol University
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SCOPUS
Bibliographic Citation
Molecular and Cellular Probes. Vol.25, No.2-3 (2011), 114-120
Suggested Citation
Nipaporn Sankuntaw, Saovaluk Sukprasert, Chulapan Engchanil, Wanlop Kaewkes, Wasun Chantratita, Vantanit Pairoj, Viraphong Lulitanond Single tube multiplex real-time PCR for the rapid detection of herpesvirus infections of the central nervous system. Molecular and Cellular Probes. Vol.25, No.2-3 (2011), 114-120. doi:10.1016/j.mcp.2011.03.004 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/11568
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Title
Single tube multiplex real-time PCR for the rapid detection of herpesvirus infections of the central nervous system
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Abstract
Human herpesvirus infection of immunocompromised hosts may lead to central nervous system (CNS) infection and diseases. In this study, a single tube multiplex real-time PCR was developed for the detection of five herpesviruses (HSV-1, HSV-2, VZV, EBV and CMV) in clinical cerebrospinal fluid (CSF) specimens. Two primer pairs specific for the herpesvirus polymerase gene and five hybridization probe pairs for the specific identification of the herpesvirus types were used in a LightCycler multiplex real-time PCR. A singleplex real-time PCR was first optimized and then applied to the multiplex real-time PCR. The singleplex and multiplex real-time PCRs showed no cross-reactivity. The sensitivity of the singleplex real-time PCR was 1 copy per reaction for each herpesvirus, while that of the multiplex real-time PCR was 1 copy per reaction for HSV-1 and VZV and 10 copies per reaction for HSV-2, EBV and CMV. Intra and inter-assay variations of the single tube multiplex assay were in the range of 0.02%-3.67% and 0.79%-4.35%, respectively. The assay was evaluated by testing 62 clinical CSF samples and was found to have equivalent sensitivity, specificity and agreement as the routine real-time PCR, but reducing time, cost and amount of used sample. © 2011 Elsevier Ltd.