Department of Microbiology, Kaohsiung Veterans General Hospital, Kaohsiung81362, Taiwan.
Department of Pediatrics, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan.
Viruses. 2018 Dec 12;10(12):711. doi: 10.3390/v10120711.
Parechovirus A (Human parechovirus, HPeV) causes symptoms ranging from severe neonatal infection to mild gastrointestinal and respiratory disease. Use of molecular approaches with RT-PCR and genotyping has improved the detection rate of HPeV. Conventional methods, such as viral culture and immunofluorescence assay, together with molecular methods facilitate comprehensive viral diagnosis. To establish the HPeV immunofluorescence assay, an antibody against HPeV capsid protein VP0 was generated by using antigenic epitope prediction data. The specificity of the anti-HPeV VP0 antibody was demonstrated on immunofluorescence assay, showing that this antibody was specific for HPeV but not enteroviruses. A total of 74 HPeV isolates, 7 non⁻polio-enteroviruses and 12 HPeV negative cell culture supernatant were used for evaluating the efficiency of the anti-HPeV VP0 antibody. The sensitivity of HPeV detection by the anti-HPeV VP0 antibody was consistent with 5'untranslated region (UTR) RT-PCR analysis. This study established comprehensive methods for HPeV detection that include viral culture and observation of cytopathic effect, immunofluorescence assay, RT-PCR and genotyping. The methods were incorporated into our routine clinical practice for viral diagnosis. In conclusion, this study established a protocol for enterovirus and HPeV virus identification that combines conventional and molecular methods and would be beneficial for HPeV diagnosis.
肠道病毒 A（人肠道病毒，HPeV）可引起从严重新生儿感染到轻微胃肠道和呼吸道疾病等各种症状。使用 RT-PCR 和基因分型等分子方法提高了 HPeV 的检测率。常规方法，如病毒培养和免疫荧光检测，与分子方法一起有助于全面的病毒诊断。为建立 HPeV 免疫荧光检测法，使用抗原表位预测数据生成了针对 HPeV 衣壳蛋白 VP0 的抗体。在免疫荧光检测中证明了抗 HPeV VP0 抗体的特异性，表明该抗体特异性针对 HPeV 而不针对肠道病毒。共使用 74 株 HPeV 分离株、7 株非脊髓灰质炎肠道病毒和 12 株 HPeV 阴性细胞培养上清液评估抗 HPeV VP0 抗体的效率。抗 HPeV VP0 抗体检测 HPeV 的敏感性与 5'非翻译区（UTR）RT-PCR 分析一致。本研究建立了包括病毒培养和观察细胞病变效应、免疫荧光检测、RT-PCR 和基因分型在内的全面 HPeV 检测方法。这些方法已纳入我们的常规临床病毒诊断实践。总之，本研究建立了一种结合常规和分子方法的肠道病毒和 HPeV 病毒鉴定方案，有助于 HPeV 诊断。
