NIAID Integrated Research Facility, Ft. Detrick, Frederick, MD, United States of America.
PLoS One. 2018 Jun 19;13(6):e0199534. doi: 10.1371/journal.pone.0199534. eCollection 2018.
Nipah virus (NiV) is a highly pathogenic zoonotic paramyxovirus that can result in severe pulmonary disease and fatal encephalitis in humans and is responsible for outbreaks in Bangladesh, Malaysia, Singapore, India and possibly the Philippines. NiV has a negative-sense RNA genome that contains six genes and serves as a template for production of viral mRNA transcripts. NiV mRNA transcripts are subsequently translated into viral proteins. Traditionally, NiV quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) assays have relied on using primer sets that amplify a target (N that encodes the nucleocapsid) within the coding region of the viral gene that also amplifies viral mRNA. Here we describe a novel one-step qRT-PCR assay targeting the intergenic region separating the viral F and G proteins, thereby eliminating amplification of the viral mRNA. This assay is more accurate than the traditional qRT-PCR in quantifying concentrations of viral genomic RNA.
尼帕病毒（NiV）是一种高致病性人畜共患病副粘病毒，可导致人类肺部严重疾病和致命脑炎，在孟加拉国、马来西亚、新加坡、印度和可能在菲律宾都有爆发。NiV 具有负义 RNA 基因组，包含六个基因，作为病毒 mRNA 转录本的模板。NiV mRNA 转录本随后被翻译为病毒蛋白。传统上，尼帕病毒实时定量逆转录聚合酶链反应（qRT-PCR）检测依赖于使用引物对扩增病毒基因编码区（编码核衣壳的 N 基因）内的靶标，该靶标也扩增病毒 mRNA。在这里，我们描述了一种针对分离病毒 F 和 G 蛋白的基因间区的新型一步 qRT-PCR 检测方法，从而消除了病毒 mRNA 的扩增。与传统 qRT-PCR 相比，该检测方法在定量病毒基因组 RNA 浓度方面更加准确。
