4.6 Article

A Novel 2-dimensional Multiplex qPCR Assay for Single-Tube Detection of Nine Human Herpesviruses

期刊

VIROLOGICA SINICA
卷 36, 期 4, 页码 746-754

出版社

KEAI PUBLISHING LTD
DOI: 10.1007/s12250-021-00354-2

关键词

Human herpesviruses (HHVs); 2-D multiplex qPCR; Melting temperature (Tm); Fluorescence probe; Melting curve

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资金

  1. National Science and Technology Major Project of China [2019YFC1200603, 2017ZX10103009-002]

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The study established a novel single-tube qPCR assay for accurate detection and differentiation of various herpesvirus infections, demonstrating high sensitivity and specificity suitable for clinical diagnosis and patient treatment.
Human herpesviruses are double-stranded DNA viruses that are classified into nine species. More than 90% of adults are ever infected with one or more herpesviruses. The symptoms of infection with different herpesviruses are diverse ranging from mild or asymptomatic infections to deadly diseases such as aggressive lymphomas and sarcomas. Timely and accurate detection of herpesvirus infection is critical for clinical management and treatment. In this study, we established a single-tube nonuple qPCR assay for detection of all nine herpesviruses using a 2-D multiplex qPCR method with a house-keeping gene as the internal control. The novel assay can detect and distinguish different herpesviruses with 30 to 300 copies per 25 mu L single-tube reaction, and does not cross-react with 20 other human viruses, including DNA and RNA viruses. The robustness of the novel assay was evaluated using 170 clinical samples. The novel assay showed a high consistency (100%) with the single qPCR assay for HHVs detection. The features of simple, rapid, high sensitivity, specificity, and low cost make this assay a high potential to be widely used in clinical diagnosis and patient treatment.

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