摘要
目的分析流行性腮腺炎病毒疫苗WM84株(以下简称WM84株)全基因序列,并与Jeryl Lynn(JL)株和腮腺炎病毒各基因型进行比对。方法 WM84株原始毒株于原代鸡胚成纤维细胞(chicken embryo fibroblast,CEF)上连续传代至7代,观察每代病毒的培养特性,测定第7代病毒全基因序列,用MEGA 4.1软件分析其核苷酸及氨基酸序列与JL株的差异,用MEGA 4.1软件的相邻连接方法(neighbor-joining,NJ)构建各基因型腮腺炎病毒的系统进化树。结果 WM84株原始病毒在CEF上连续传7代的各代病毒的培养特性基本稳定。WM84株与JL2及JL5株比较,核苷酸序列同源性分别为99.90%和97.17%。WM84株与JL2株比较,核衣壳蛋白(N)、磷酸化蛋白(P)、基质蛋白(M)、融合蛋白(F)、小疏水蛋白(SH)、血凝素-神经氨酸酶蛋白(HN)、大蛋白(L)核苷酸序列同源性分别为为99.94%、99.92%、99.91%、99.81%、98.84%、99.94%、100%;氨基酸序列的同源性分别为100%、100%、99.73%、99.44%、98.23%、100%、100%。WM84株与JL5株比较,N、P、M、F、SH、HN、L蛋白核苷酸序列同源性分别为97.59%、96.84%、97.93%、97.02%、94.01%、97.02%、97.63%;氨基酸序列的同源性分别为98.72%、94.27%、98.39%、97.36%、90.82%、97.57%、99.42%。WM84株与JL2、JL5株SH蛋白的第29位和第48位氨基酸位点上无差异。WM84、JL2和JL5株均为A基因型,WM84株与JL2株的遗传距离低于与JL5株的遗传距离。结论 WM84株CEF 7代病毒与JL2株的同源性较高,其致病性、中和活性相关位点均未发生改变,表明WM84株具有稳定的免疫原性,其制备的疫苗安全有效。
Objective To sequence and analyze the complete gene of mumps vaccine virus WM84 strain,and compare the result with those of Jeryl Lynn（JL） strain and munps virus of various genotypes.Methods WM84 strain was subcultured in primary chicken embryo fibroblasts（CEFs） to passage 7,and the cultural characteristics of each passage was observed.The complete gene of passage 7 was sequenced,while the difference of nucleotide and amino acid sequences from those of JL strain were analyzed by using MEGA 4.1 software,based on which a phylogenetic tree of mumps virus of various genotypes was constructed by the neighbor-joining（NJ） method in the software.Results The cultural characteristics and virus titer of WM84 strain of various passages were basically stable.The homologies of nucleotide sequences of WM84 strain to those of JL2 and JL5strains were 99.90% and 97.17% respectively.The homologies of nucleotide sequences of nucleocapsid（N）,phosphorylated（P）,matrix（M）,fusion（F）,small hydrophobic（SH）,hemagglutinin-neuraminidase（HN）,and large（L）proteins of WM84 strain were 99.94%,99.92%,99.91%,99.81%,98.84%,99.94% and 100%,while those of amino acid sequences were 100%,100%,99.73%,99.44%,98.23%,100% and 100%,to those of JL2 strain,respectively.However,the homologies of nucleotide sequences of N,P,M,F,SH,HN and L proteins of WM84 strain were 97.59%,96.84%,97.93%,97.02%,94.01%,97.02% and 97.63%,while those of amino acid sequences were 98.72%,94.27%,98.39%,97.36%,90.82%,97.57% and 99.42%,to those of JL5 strain,respectively.The SH protein at sites 29 and 48 of WM84 strain showed no significant difference with those of JL2 and JL5strains.WM84,JL2 and JL5strains were of genotype A.The genetic distance of WM84 strain to JL2 strain was closer than that to JL5 strain.Conclusion The passage 7 of WM84 strain showed high homology to JL2 strain,of which the sites associated with pathogenicity and neutralizing activity showed no change,indicating stable immunogenicity.The vaccine prepared with the strain was safe and effective.
出处
《中国生物制品学杂志》
CAS
CSCD
2015年第11期1113-1117,1126,共6页
Chinese Journal of Biologicals
基金
国家科技重大专项"重大新药创制"课题(2013ZX09402302)
作者简介
通讯作者：魏树源，E-mail：weishuyuan@sinopharm．com
