Journal
DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE
Volume 93, Issue 2, Pages 101-106Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.diagmicrobio.2018.09.001
Keywords
Respiratory syncytial virus (RSV); Viral detection; Locked nucleic acid (LNA); One-tube nested real time RT-PCR (OTNRT-PCR); Melting curve analysis
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Funding
- National Key Research and Development Plan of China [2016YFC1202700, 2016YFC1200903, 2017YFC1200503]
- China Mega-Project for Infectious Disease [2017ZX10302301-004, 2017ZX10104001]
- Hebei Key Project Plan for Medical Science Research [20180616]
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Respiratory syncytial virus (RSV) causes serious respiratory tract infection worldwide. The relatively low RSV load makes it difficult to detect in frail, elderly, and severely immune-compromised patients. In the present study, we developed a locked nucleic acid-based 1-tube nested real-time RT-PCR (OTNRT-PCR) assay with the advantages of extremely high sensitivity, facile operability, and less likelihood of cross-contamination. The sensitivity, specificity, and clinical performance of the OTNRT-PCR assay were compared in parallel with a conventional TaqMan probe-based real-time PCR (qRT-PCR) assay and a traditional 2-step nested RT-PCR assay. The limit of detection of the OTNRT-PCR assay was 1.02 x 10(-1) TCID50/mL, equivalent to the traditional 2-step nested RT-PCR assay and 25-fold lower than the qRT-PCR assay. Of 616 nasopharyngeal aspirates tested, 143 RSV-negative samples by qRT-PCR were confirmed as positive by sequencing the OTNRT-PCR products. We therefore conclude that OTNRT-PCR is more sensitive than qRT-PCR for detection of RSV in clinical samples. (C) 2018 Elsevier Inc. All rights reserved.
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