Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, Pohang, 790-784, Republic of Korea.
R&D Center, Ahram Biosystems Inc, Seoul, 133-120, Republic of Korea.
Mol Biol Rep. 2019 Oct;46(5):5073-5077. doi: 10.1007/s11033-019-04961-x. Epub 2019 Jul 12.
One of the most crucial steps for preventing viral pandemics is the early detection of the causative virus on site. Various molecular and immunological approaches have been developed for virus detection. In this study, we investigated the utility of the recently introduced convection polymerase chain reaction (cPCR) platform for the rapid and sensitive detection of various animal viruses in the field, including the foot-and-mouth disease virus (FMDV) and avian influenza viruses (AIVs). Primer sets were designed to simultaneously detect two highly conserved regions of the FMDV, including the 5' untranslated region (5'-UTR) and 3D gene, and to specifically amplify the NP and hemagglutinin (HA) genes of H5 and H9 subtypes of AIVs. The portable cPCR system was able to amplify from as low as 1 to 10 copies of viral cDNAs in the singleplex mode and 10 to 100 copies of viral cDNAs in the duplex mode within 21 min. Thus, our data suggest that the cPCR protocols developed in this study are highly sensitive and enable quick detection of animal viruses in biological samples.
预防病毒大流行的最关键步骤之一是在现场尽早检测到致病病毒。已经开发了各种分子和免疫学方法来检测病毒。在这项研究中，我们研究了最近引入的对流聚合酶链反应（cPCR）平台在现场快速灵敏地检测各种动物病毒的效用，包括口蹄疫病毒（FMDV）和禽流感病毒（AIVs）。设计了引物组以同时检测 FMDV 的两个高度保守区域，包括 5'非翻译区（5'-UTR）和 3D 基因，并特异性扩增 H5 和 H9 亚型 AIV 的 NP 和血凝素（HA）基因。便携式 cPCR 系统能够在单重模式下从低至 1 到 10 个拷贝的病毒 cDNA 扩增，在双重模式下从 10 到 100 个拷贝的病毒 cDNA 扩增，时间在 21 分钟内。因此，我们的数据表明，本研究中开发的 cPCR 方案具有高度的敏感性，能够快速检测生物样本中的动物病毒。
