Simultaneous detection and differentiation of human parvovirus B19 and human parvovirus 4 by an internally controlled multiplex quantitative real-time PCR
Simultaneous detection and differentiation of human parvovirus B19 and human parvovirus 4 by an internally controlled multiplex quantitative real-time PCR
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通过内控多重实时 PCR 同时检测和区分人细小病毒 B19 和人细小病毒 4
DOI：
10.1016/j.mcp.2017.08.005
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发表时间：
2017-12-01
影响因子：
3.3
通讯作者：
Zhang, Jingang
中科院分区：
文献类型：
--
作者：
Jia, Junting;Zhong, Yadi;Zhang, Jingang
人细小病毒B19（B19 V）和人细小病毒4（PARV 4）是已知可感染人类并通过血液和血浆源性药品（PDMP）传播的两种细小病毒。灭活这两种细小病毒已被证明是困难的和核酸检测（NAT）。是一种有效的排除病毒的方法。本研究建立了B19 V和PARV 4同时检测和定量的内部控制多重实时定量PCR（qPCR）方法，并对其进行了评价。优化的多重qPCR检测允许同时检测B19 V和PARV 4的所有基因型（1-3），定量限（LOQ）相同，为5拷贝/μ L，而不是其他血液传播病毒。它具有至少8个数量级的可靠放大线性的宽动态范围。B19 V和PARV 4的定量循环（Cq）值的低标准偏差（SD）和拷贝数的低变异系数（CV）表明多重qPCR检测具有高水平的重复性和再现性。该多重qPCR检测可作为一种易于适用的方法，用于筛选预期进一步生产成PDMP的血浆单位，以降低此类产品感染细小病毒的风险，并且还可用于检测B19 V/PARV 4共感染或共存。(C)2017爱思唯尔有限公司版权所有。
Human parvovirus B19 (B19V) and human parvovirus 4 (PARV4) are two parvoviruses known to infect humans and transmit through blood and plasma derived medicinal products (PDMPs). Inactivation of the two parvoviruses has proven to be difficult and nucleic acid testing (NAT). would be an efficient means to exclude viruses. In this study, an internally controlled multiplex quantitative real-time PCR (qPCR) assay for B19V and PARV4 simultaneous detection and quantification was established and evaluated. The optimized multiplex qPCR assay allowed for simultaneous detection of all of the genotypes (1-3) of B19V and PARV4, with equal limit of quantification (LOQ) of 5 copies/mu L, rather than other blood-borne viruses. It had a wide dynamic range of reliable amplification linearity of at least 8 orders of magnitude. Low standard deviations (SD) of quantification cycle (Cq) values and low coefficients of variation (CV) of copy numbers for both B19V and PARV4 suggested a high level of repeatability and reproducibility for the multiplex qPCR assay. This multiplex qPCR assay can be served as a readily applicable approach to screen plasma units intended for further manufacturing into PDMPs to reduce the risk of parvoviruses infection by such products and may also be useful for the detection of B19V/PARV4 co-infection or co-existence. (C) 2017 Elsevier Ltd. All rights reserved.