本文選題：手足口病 + EV�。� 參考：《中國病原生物學(xué)雜志》2017年12期

【摘要】：目的了解淮南市手足口病病毒分離株VP1基因遺傳特征及進(jìn)化關(guān)系。方法用人橫紋肌肉瘤(rhabdomyosarcoma,RD)細(xì)胞對2014-2016年熒光PCR檢測為EV71型、CA16型及CA10型陽性標(biāo)本進(jìn)行病毒分離,經(jīng)2代接種出現(xiàn)細(xì)胞病變(CPE)后取培養(yǎng)物上清,提取病毒RNA,RT-PCR法擴(kuò)增不同基因型VP1基因并測序,采用Bioedit5.0.6、DNAstar7.1和Mega 6.06生物軟件對測序結(jié)果進(jìn)行比對及進(jìn)化分析。結(jié)果 11株EV71型分離株VP1編碼區(qū)的核苷酸片段長度為891nt,同源性為93.5%-100.0%,氨基酸同源性為98.3%-100.0%;7株CVA16型分離株VP1編碼區(qū)的核苷酸片段長度為891nt,同源性為95.1%-97.0%,氨基酸同源性為98.7%-100.0%;3株CVA10型分離株VP1編碼區(qū)的核苷酸片段長度為894nt,同源性為96.5%-100.0%,氨基酸同源性則為98.7%-100.0%。系統(tǒng)進(jìn)化分析EV71型淮南株與安徽阜陽、山東、河南、北京、云南分離毒株親緣性較近,處于進(jìn)化分支C4a,核苷酸同源性為93.5%-100.0%,氨基酸同源性為98.7%-100.0%;CVA16型淮南株為B1b亞型,與山東株、河南株、深圳株、南京株親緣關(guān)系較近,核苷酸序列同源性為95.1%-99.1%,氨基酸序列同源性98.7%-100.0%;CVA10淮南株處于D分支內(nèi),與云南、河南、河北株親緣關(guān)系較近,核苷酸序列同源性為96.5%-100.0%,氨基酸序列同源性98.3%-100.0%。結(jié)論淮南市2014-2016年流行的EV71為C4a亞型,CVA16為B1b亞型,CVA10處于D聚類分支,與國內(nèi)其他地區(qū)流行的手足口病毒亞型一致。
[Abstract]:Objective to investigate the genetic characteristics and evolutionary relationship of VP1 gene of HFMD virus isolate in Huainan City. Methods Human rhabdomyosarcomac Rd cells were isolated from positive specimens of EV71 type CA16 and CA10 detected by fluorescent PCR from 2014-2016. The supernatant of culture was obtained after inoculation with cytopathic acid (CPE) for 2 passages. The VP1 gene of different genotypes was amplified by RT-PCR and sequenced. Bioedit5.0.6 DNA star7.1 and Mega 6.06 were used to compare and analyze the sequencing results. Results the nucleotide fragment length of VP1 coding region in 11 EV71 isolates was 891nt.The homology was 93.5-100.0.The amino acid homology was 98.3-100.00.The nucleotide length of VP1 coding region of 7 CVA16 isolates was 891nt, the homology was 95.1 -97.0, and the amino acid homology was 98.7- 100.0.The amino acid homology was 98.7- 100.020. The nucleotide fragment length of VP1 coding region of CVA10 isolate was 894nt.The homology was 96.5- 100.0.The amino acid homology was 98.7- 100.0. Phylogenetic analysis showed that Huainan strain EV71 was closely related to the isolates isolated from Fuyang, Shandong, Henan, Beijing and Yunnan, and was in the evolutionary branch of C4a. The nucleotide homology was 93.5-100.0.The amino acid homology was 98.7-100.0B1b subtype of Huainan strain of CVA16, which was subtype of B1b with Shandong strain. The homology of nucleotide sequence of Henan strain, Shenzhen strain and Nanjing strain was 95.1 -99.1, the amino acid sequence homology was 98.7-100.00.The Huainan strain of CVA10 was in D branch, and had close relationship with Yunnan and Hebei strains. The homology of nucleotide sequence was 96.5- 100.0.The homology of amino acid sequence was 98.3- 100.0km. Conclusion the EV71 epidemic in Huainan City from 2014-2016 is C4a subtype, CVA16 is B1b subtype, CVA10 is in D cluster branch, which is consistent with the HFMD virus subtype prevalent in other parts of China.
【作者單位】： 淮南市疾病預(yù)防控制中心;安徽省疾病預(yù)防控制中心;
【基金】：淮南市市級科技計劃項目(No.1307)
【分類號】：R440;R725.1

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