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目的鉴定L20B细胞[转人脊髓灰质炎(脊灰)病毒受体基因的小鼠肺细胞系]支持生长的非脊灰肠道病毒(Non-polio Enteroviruses,NPEV)的血清型,并分析病毒基因特征,探讨其对脊灰病毒(Poliovirus,PV)分离率的影响。方法收集宁夏回族自治区从健康人群和广西壮族自治区从急性弛缓性麻痹病例粪便标本中分离的5株L20B细胞支持生长的NPEVs,对病毒分离物采用逆转录-聚合酶链反应进行VP4编码区基因扩增和核苷酸序列测定和分析,根据序列比对结果确定血清型,再进行全长VP1编码区序列测定和分析,并和基因数据库下载的相应血清型NPEVs的VP1编码区核苷酸序列进行比对和分析,构建基因亲缘性关系树。结果 5株NPEVs均鉴定为柯萨奇A组病毒(Coxsackie Virus GroupA,CVA),其中1株为CVA4,另1株为CVA8,其余3株均为CVA10,均属A组人肠道病毒(Human Enteroviruses,HEV);通过比较全长VP1编码区核苷酸同源性和构建亲缘关系树发现,3个血清型的中国分离株都分别独立成簇,与其他国家分离的相应血清型的NPEVs及原型株病毒差异都较大,是本土流行病毒株。结论所研究的L20B细胞支持生长的5株NPEVs均属A组HEV,包括CVA4、CVA8、CVA10三个血清型。由于L20B细胞对PV的最初接种具有很高的敏感性,因此L20B细胞即使支持某些CVA生长,也不会影响PV分离率。但在近年来中国发生较大规模手足口病(Hand,Foot and Mouth Disease,HFMD)的背景下,由于这些A组HEV同时是HFMD的病原体,对这些病毒的鉴定对PV的鉴别诊断和HFMD的预防控制都有实际意义。
Objective To identify the serotypes of non-polio Enteroviruses (NPEV) that support growth of L20B cells [mouse lung cell line transfected with poliovirus (poliovirus) receptor gene] and to analyze the viral genes Characteristics, to explore its poliovirus (Poliovirus, PV) separation rate. Methods Five NPEVs isolated from stool specimens from healthy stool samples of Guangxi Zhuang Autonomous Region isolated from stool samples of acute flaccid paralysis were collected and the virus isolates were amplified by reverse transcription polymerase chain reaction The nucleotide sequence was determined and analyzed. The serotypes were determined according to the alignment results. The full-length VP1 coding region was sequenced and analyzed. The nucleotide sequence of the VP1 coding region of the corresponding serotype NPEVs was downloaded from the GenBank Alignment and analysis, construction of gene relationship tree. Results All five NPEVs were identified as Coxsackie Virus Group A (CVA), of which one was CVA4 and the other was CVA8. The remaining three were all CVA10, which belonged to group A human enterovirus Enteroviruses, HEV). Comparing the nucleotide homology of the full-length VP1 coding region with the constructed phylogenetic tree, it was found that all three isolates of Chinese serotypes were clustered separately and NPEVs of corresponding serotypes separated from other countries Prototype strains of viruses are larger, is a native strain of endemic. Conclusions All the 5 NPEVs supported by L20B cells are group A HEV, including three serotypes of CVA4, CVA8 and CVA10. Because L20B cells are highly sensitive to the initial inoculation of PV, L20B cells do not affect PV isolation even if they support some CVA growth. However, in the context of the large-scale Hand, Foot and Mouth Disease (HFMD) in China in recent years, since these group A HEVs are both pathogens of HFMD, the differential diagnosis of PV and the differential diagnosis of HFMD Prevention and control have practical significance.