Dimitriou T G, Pliaka V, Kyriakopoulou Z, Ruether I G A, Tsakogiannis D, Fountoucidou P, Gartzonika C, Levidiotou-Stefanou S, Markoulatos P
University of Thessaly, School of Health Sciences, Department of Biochemistry & Biotechnology, Microbiology-Virology Laboratory, Larissa, Greece.
University of Ioannina, Medical School, Department of Microbiology, Ioannina, Greece.
Mol Cell Probes. 2014 Aug;28(4):107-12. doi: 10.1016/j.mcp.2013.10.004. Epub 2013 Dec 4.
Poliomyelitis has been effectively controlled by the use of inactivated poliovirus vaccine (IPV) or trivalent live attenuated oral poliovirus vaccine (OPV). Since 1964, the use of OPV in mass vaccinations has resulted in drastic reductions of the number of poliomyelitis cases caused by wild-type polioviruses. However, the characterization of OPV derivatives with increased neurovirulence, constituted a real problem with respect to OPV safety. Mutations at attenuating sites of the genome and recombination events between Sabin strains of the trivalent OPV vaccine have been correlated with the loss of the attenuated phenotype of OPV strains and the acquisition of traits characteristic of wild polioviruses. In consequence, early detection and characterization of recombinant evolved derivatives of vaccine strains is highly important. In this report, ten PCR assays are described which allow for the identification of rare recombination events located in VP1, 2A, 2C, 3A, 3C and 3D genomic regions and predominant recombination events located in 2C and 3D genomic regions of OPV derivatives. These assays could be readily implemented in diagnostics laboratories lacking sequencing facilities as a first approach for the early detection and characterization of recombinant OPV derivatives.
脊髓灰质炎已通过使用灭活脊髓灰质炎病毒疫苗(IPV)或三价口服减毒活脊髓灰质炎病毒疫苗(OPV)得到有效控制。自1964年以来,在大规模疫苗接种中使用OPV已使野生型脊髓灰质炎病毒引起的脊髓灰质炎病例数大幅减少。然而,神经毒性增加的OPV衍生物的特性,构成了OPV安全性方面的一个实际问题。三价OPV疫苗的Sabin株基因组减毒位点的突变以及重组事件,与OPV株减毒表型的丧失和野生脊髓灰质炎病毒特征性状的获得有关。因此,早期检测和鉴定疫苗株的重组进化衍生物非常重要。在本报告中,描述了十种PCR检测方法,这些方法可用于鉴定位于OPV衍生物的VP1、2A、2C、3A、3C和3D基因组区域的罕见重组事件,以及位于2C和3D基因组区域的主要重组事件。在缺乏测序设施的诊断实验室中,这些检测方法可作为早期检测和鉴定重组OPV衍生物的首选方法轻松实施。