2Laboratório de Vírus Respiratório e do Sarampo, Pavilhão Helio Peggy e Pereira, IOC, Fiocruz, Av. Brasil 4365, Manguinhos, Rio de Janeiro/RJ, Brazil.
3National Institute for Science and Technology on Innovation on Diseases of Neglected Populations (INCT/IDPN), Center for Technological Development in Health (CDTS), Fiocruz, Av. Brasil 4036, Manguinhos, Rio de Janeiro/RJ, Brazil.
J Gen Virol. 2018 Dec;99(12):1608-1613. doi: 10.1099/jgv.0.001168. Epub 2018 Nov 5.
Influenza A and B virions are packaged with their polymerases to catalyse RNA-dependent RNA polymerase activity. Since there is no evidence to rule in or out the permissiveness of influenza virions to triphosphate ribonucleotides, we functionally evaluated this. We found the means to stimulate influenza A and B RNA polymerase activity inside the virion, called natural endogenous RNA polymerase (NERP) activity. Stimulation of NERP activity increased up to 3 log10 viral RNA content, allowing the detection of influenza virus in otherwise undetectable clinical samples. NERP activation also improved our capacity to sequence misidentified regions of the influenza genome from clinical samples. By treating the samples with the ribavirin triphosphate we inhibited NERP activity, which confirms our hypothesis and highlights that this assay could be used to screen antiviral drugs. Altogether, our data show that NERP activity could be explored to increase molecular diagnostic sensitivity and/or to develop antiviral screening assays.
甲型和乙型流感病毒衣壳与聚合酶一起包装,以催化 RNA 依赖的 RNA 聚合酶活性。由于没有证据表明流感病毒衣壳对三磷酸核糖核苷酸具有允许性或不允许性,因此我们对此进行了功能评估。我们发现了一种在病毒衣壳内刺激甲型和乙型流感 RNA 聚合酶活性的方法,称为天然内源性 RNA 聚合酶(NERP)活性。NERP 活性的刺激可使病毒 RNA 含量增加多达 3 个对数,从而使原本无法检测到的临床样本能够检测到流感病毒。NERP 激活还提高了我们从临床样本中正确识别流感基因组区域的能力。通过用利巴韦林三磷酸处理样品,我们抑制了 NERP 活性,这证实了我们的假设,并强调了该测定可用于筛选抗病毒药物。总之,我们的数据表明,可以探索 NERP 活性来提高分子诊断的敏感性和/或开发抗病毒筛选测定。