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.

Nipah病毒（NiV）是一种高致病性人畜共患副粘病毒，可导致人类严重的肺部疾病和致命性脑炎，并导致孟加拉国，马来西亚，新加坡，印度以及菲律宾爆发疫情。NiV具有一个包含六个基因的负义RNA基因组，可作为生产病毒mRNA转录本的模板。NiV mRNA转录物随后被翻译成病毒蛋白。传统上，NiV定量实时逆转录酶聚合酶链反应（qRT-PCR）分析依赖于使用引物组，该引物组在还扩增病毒mRNA的病毒基因的编码区域内扩增靶标（编码核衣壳的N）。在这里，我们描述了一种针对分离病毒F和G蛋白的基因间区域的新颖的一步式qRT-PCR检测方法，从而消除了病毒mRNA的扩增。此方法在定量病毒基因组RNA浓度方面比传统的qRT-PCR更准确。