猪伪狂犬病毒Jiangxi-FZ株的分离鉴定及其重要毒力基因分子特征北大核心CSCD

目的研究伪狂犬病病毒(PRV)新流行毒株重要毒力基因的分子特征。方法本研究从江西省某规模化猪场疑似猪伪狂犬病发病仔猪的脑组织中分离到1株病毒,通过PCR鉴定、病毒分离培养、细胞免疫荧光及易感动物试验证实该病毒为PRV野毒株并命名为PRV Jiangxi-FZ株。并对其重要毒力基因TK、gB、gC、gD及gE的分子特征和遗传进化关系进行分析。结果 Jiangxi-FZ株与其他PRV参考毒株的TK、gB、gC、gD及gE基因在核苷酸和氨基酸水平均具有很高的保守性,尤其是与2012年分离的2株PRV变异株的同源性较高。但在高度保守的基础上,仍存在一些差异,且部分差异具有特征性。遗传进化树分析结果显示,PRV Jiangxi-FZ株与2012年国内不同省份分离的PRV变异株亲缘关系较近,而与Becker等欧美洲毒株亲缘关系相对较远。结论 Jiangxi-FZ株具有当前PRV流行毒株的代表性,属近年来流行的PRV变异毒株。     

虎源伪狂犬病毒的鉴定及gB基因分析

目的对不明原因死亡虎进行病毒感染检测,并对病毒部分保守基因序列进行分析。方法采集不明原因病死虎肺组织,研磨后提取总DNA,使用特异性引物对伪狂犬病毒gB基因、gD基因进行PCR扩增、测序并构建进化树;经负染法染色后用电子显微镜观察病毒形态结构;取研磨组织液接种家兔,观察病毒的致病力。结果 PCR扩增组织样品为gB基因、gD基因阳性,基因序列经blast比对伪狂犬病毒。将gB基因通过序列测定分析并与GenBank中近4年公布的9株伪狂犬病毒gB基因比对,同源性为99.5%~100%。进化树分析该病毒与2012年北京家猪分离株属同一进化分支。组织研磨液置电镜观察,见病毒呈圆形,直径100nm左右,有囊膜,为具有典型特征的疱疹病毒样病毒粒子。家兔接种组织研磨液后出现啃咬接种部位,抽搐,嘶叫等伪狂犬病症状。结论本研究在病死虎体内检测到伪狂犬病毒,表明濒危野生动物虎已受到伪狂犬病毒病的威胁。     

两株犬源狂犬病病毒街毒株全基因组序列测定与分析

目的对2株犬源狂犬病病毒街毒株全基因组测序,了解国内狂犬病病毒的流行和变异情况。方法采用乳鼠脑内接种方法分离2株天津地区狂犬病病毒街毒株,RT-PCR扩增病毒DNA覆盖全基因组12个相互重叠的基因片段,PCR产物平端克隆后进行全基因组核苷酸序列测定,然后对其分子变异和进化特点进行分析。进行两分离株的序列相似性、氨基酸同源性、主要功能位点的变异比较和种系发生分析。结果 TJD04和TJD05两株狂犬病病毒全基因组长均为11 924 nts,全基因组序列的组成和结构均符合弹状病毒科狂犬病病毒属的特征。病毒基因组由5个编码区组成,基因起始位点和终止位点高度保守,氨基酸编码长度未发生变异,5个编码蛋白序列较少发生变化,仅有个别主要功能位点发生变异,且大部分变异均属于无意义沉默突变。多序列相似性比较和种系发生分析显示,两株病毒均属于基因1型,N基因与全基因进化树保持一致,与中国犬源狂犬病病毒BD06同源性最高(99.6%),进化关系近,并具有较为独特的中国地域特点。结论两天津株狂犬病毒可能是自然界中固有的狂犬病病毒街毒株。     

我国分离的10株狂犬病病毒P和M蛋白基因序列分析

摘 要：目的 探讨我国狂犬病病毒P、M蛋白基因序列、结构特点及变异情况等遗传学特征。 方法 用RT-PCR方法从分离的10株狂犬病病毒中获得目的基因片段,测定核苷酸序列后,利用生物信息学软件分析核苷酸、氨基酸序列及其相关的功能位点并构建P、M蛋白基因的系统发育树。结果 10株病毒P和M基因核苷酸序列同源性分别为85.9％～99.4％和89.5％～99.5％,推导出的氨基酸同源性分别为92.3％～100％和96.0％～99.5％。在核苷酸及氨基酸水平上,10株病毒与我国分离的街毒株HN10及CTN疫苗株的P、M基因同源性均明显高于国外其它疫苗株、标准攻击毒CVS株相应的同源性。系统发育分析表明,10株病毒与我国湖南街毒株、CTN疫苗株进化关系最近,而与研究中选取的其他毒株进化关系较远。氨基酸对位分析表明,与其它基因1型毒株相比,本研究中10株狂犬病病毒的P、M基因出现多处变异,但很少在功能区发生变异。结论 分离的10株病毒属基因l型狂犬病病毒,与我国分离的街毒株及CTN疫苗株关系较近。     

云南省通海县蚊虫2株乙型脑炎病毒的分离鉴定北大核心CSCD

目的了解云南省通海县蚊虫携带的乙型脑炎病毒(JEV)情况、分子特征及基因分型。方法2015年7月在云南省玉溪市通海县采用诱蚊灯在调查点通宵捕捉蚊虫,蚊虫研磨后分别接种BHK-21和C6/36细胞进行病毒分离,使用黄病毒属特异引物进行初步鉴定,使用乙型脑炎病毒E基因特异引物进行RT-PCR扩增及测序,使用Megalign、Genedoc、Mega7等软件进行序列分析。结果共捕获三带喙库蚊、中华按蚊、致倦库蚊和骚扰阿蚊2300只,其中三带喙库蚊为优势蚊种(78%)。分46批进行病毒分离,获得2株阳性分离物,接种BHK-21细胞96h,引起细胞病变。使用黄病毒属特异引物和乙型脑炎病毒E基因特异引物扩增均为阳性。序列分析结果显示,2株新分离乙型脑炎病毒与疫苗株SA14-14-2在E基因存在12个氨基酸差异位点,在8个与乙脑病毒毒力相关的位点上存在6个差异位点。E基因遗传进化分析显示,2株新分离病毒与基因Ⅰ型乙型脑炎病毒位于同一进化分支,核苷酸和氨基酸同源性分别为91.8%~99.1%和98.0%~100%。遗传进化分析显示,其与云南2016年蚊虫中分离的乙脑病毒株JEV/mosq/YN/2016亲缘关系较近,核苷酸和氨基酸同源性分别为99.1%和99.8%。结论云南省通海县分离的2株病毒鉴定为基因Ⅰ型乙型脑炎病毒,其与云南省近年来流行的毒株遗传进化关系较近。2毒株的E基因上与毒力相关的关键位点中有2个位点发生突变,结构域Ⅲ的关键抗原未发生改变,因此SA14-14-2减毒疫苗株仍可用于该地区人群的免疫接种,以预防乙型脑炎的发生和流行。     

云南省通海县蚊虫2株乙型脑炎病毒的分离鉴定

目的了解云南省通海县蚊虫携带的乙型脑炎病毒(JEV)情况、分子特征及基因分型。方法 2015年7月在云南省玉溪市通海县采用诱蚊灯在调查点通宵捕捉蚊虫,蚊虫研磨后分别接种BHK-21和C6/36细胞进行病毒分离,使用黄病毒属特异引物进行初步鉴定,使用乙型脑炎病毒E基因特异引物进行RT-PCR扩增及测序,使用Megalign、Genedoc、Mega7等软件进行序列分析。结果共捕获三带喙库蚊、中华按蚊、致倦库蚊和骚扰阿蚊2 300只,其中三带喙库蚊为优势蚊种(78%)。分46批进行病毒分离,获得2株阳性分离物,接种BHK-21细胞96 h,引起细胞病变。使用黄病毒属特异引物和乙型脑炎病毒E基因特异引物扩增均为阳性。序列分析结果显示,2株新分离乙型脑炎病毒与疫苗株SA14-14-2在E基因存在12个氨基酸差异位点,在8个与乙脑病毒毒力相关的位点上存在6个差异位点。E基因遗传进化分析显示,2株新分离病毒与基因Ⅰ型乙型脑炎病毒位于同一进化分支,核苷酸和氨基酸同源性分别为91.8%～99.1%和98.0%～100%。遗传进化分析显示,其与云南2016年蚊虫中分离的乙脑病毒株JEV/mosq/YN/2016亲缘关系较近,核苷酸和氨基酸同源性分别为99.1%和99.8%。结论云南省通海县分离的2株病毒鉴定为基因Ⅰ型乙型脑炎病毒,其与云南省近年来流行的毒株遗传进化关系较近。2毒株的E基因上与毒力相关的关键位点中有2个位点发生突变,结构域Ⅲ的关键抗原未发生改变,因此SA14-14-2减毒疫苗株仍可用于该地区人群的免疫接种,以预防乙型脑炎的发生和流行。     

2株欧亚与北美谱系重排H6N8亚型禽流感病毒的遗传进化及分子特征分析北大核心CSCD

目的解析监测中发现的欧亚与北美谱系重组H6N8亚型禽流感病毒的遗传进化及分子特征,为禽流感病毒跨洲际传播及基因重排研究提供支持。方法2019年秋季在图牧吉地区采集雁鸭类粪便样品,分离H6N8亚型禽流感病毒并进行全基因序列测定,分析其遗传进化及分子特征。结果共采集雁鸭类粪便及拭子样品5062份,分离到2株H6N8LPAIV(TMJ43、TMJ120)。遗传进化分析显示,分离株病毒NA与北美的阿拉斯加地区所分离的H3N8亚型AIV的NA节片聚集在同一进化分支,HA和其它6个内部基因节片均属于欧亚进化分支,且两株禽流感病毒分别有不同的重配发生。2株H6N8亚型禽流感病毒为欧亚谱系与北美谱系基因重排毒株,其中N8基因最近一次欧亚与北美基因节片重排发生在2016年。分子特征分析显示,HA蛋白的碱性裂解位点为PQIETR↓GLF,符合低致病性禽流感病毒的特征。在NP蛋白,发生了具有增强鸡禽流感毒力的A184K突变。在TMJ43,NS1蛋白发生了能增强哺乳动物毒力的A/P42S突变。结论在我国内蒙古地区分离的2株H6N8亚型禽流感病毒为欧亚谱系和北美谱系的重排毒株,两毒株均发生了增强家禽和哺乳动物感染能力的突变。表明北美毒株通过候鸟迁徙进入我国,与欧亚谱系毒株发生重配并在野鸟中频繁出现,可为禽流感病毒的跨洲际传播研究提供信息支持。     

源于亲本株PRVFa的3个基因缺失病毒株PRV TK^-、PRV gE^-／gI^-和PRV TK^-／gE^-／gI^-抵抗强毒攻击的比较研究

目的 4周龄仔猪24头随机分成4个组(A、B、C和对照组),前3组中各5头分别对应接种对应的缺失毒力基因TK(A)、缺失毒力基因gE/gI(B)和缺失毒力基因TK/gE/gI(C)3个基因缺失株,接种剂量均为105PFU,留1头不接种作为同居猪.免疫14d后以107PFU PRV Fa株滴鼻攻毒所有试验猪, 7d后屠宰猪只,采集大脑、小脑、心脏、肝脏、肺脏、脾脏、肾脏、扁桃体、下颌淋巴结和三叉神经节,用光镜和电镜技术观察PRV攻毒后仔猪的器官损伤程度.收集免疫接种后3d内和强毒攻击后5d内的所有猪只鼻腔拭纸,以Southern blots方法测定PRV 的组织分布情况.结果 表明,收集的10种组织中出现病变的组织比例分别为:对照组为9/10,免疫A组为 4/10、B组为3/10、C组为4/10.病理学和超微病例观察表明,对照组和A组中肺脏损伤严重,其他组轻微;对照组中大脑、小脑、三叉神经损伤严重,免疫组轻微;A、B和C疫苗株对潜伏感染的主要靶器官扁桃体损伤程度依次加重.Southern blots分析表明,所有接种缺失病毒的试验猪均可通过鼻腔分泌物散毒,但是排出的病毒不能成功感染同居猪;A、B和C疫苗株免疫后均不能有效阻止PRV Fa 攻击后的散毒,但在仔猪大脑和小脑中不能检出病毒.结论 接种3个不同基因缺失株A、B和C均能阻止强毒株Fa 感染后向大脑和小脑的入侵,并减少强毒对多个器官的损害作用,其中C株的抗强毒感染能力和抗强毒所致损伤能力更佳.     

2016-2018年广州市H6亚型禽流感病毒外环境分离株HA基因遗传特征分析

目的 对广州禽类市场外环境H6亚型禽流感病毒进行分离和测序,分析HA基因遗传进化特点。方法 通过接种鸡胚分离H6亚型禽流感病毒,采用RT-PCR方法扩增HA基因全长序列并测序,通过DNA Star7.1和MEGA 4.0软件,分析HA基因遗传特征。结果 2016-2018年广州禽类市场外环境分离到6株H6亚型禽流感病毒,HA基因核苷酸同源性在81.0%~99.1%之间。基因序列特征分析显示,所有分离株裂解位点序列相对保守,只有1个碱性氨基酸插入,呈现低致病性禽流感病毒分子特点。受体结合位点均为226Q和228G,为禽类呼吸道上皮受体结合位点。2016年分离株发现183-NNT糖基化位点的缺失。进化分析显示,广州早期分离株属于广东常见的ST2853-like分支,但2018年监测发现了ST339-like新流行分支病毒。结论 广州地区H6亚型禽流感病毒尚为禽类低致病性病毒,本研究毒株HA基因变异较大, 2018年毒株出现多遗传分支进化趋势,因此需要继续监测H6亚型禽流感病毒的流行和变异。     

重庆地区2004年12月～2005年4月犬狂犬病流行毒株的检测

目的 确诊重庆地区2004年12月至2005年4月发生的临床疑似犬狂犬病，分离病毒并分析病毒株的进化关系。方法 RT，PCR检测重庆地区的5例临床疑似狂犬脑组织样品，乳鼠脑内接种试验分离病毒，分析5个流行毒株间及其与占各基因型代表毒株的遗传衍化关系。结果 5例临床样品均含有狂犬病毒，分离获得了相应的5株流行毒，CQ／ws-1，aQ／wx-1，aQ／w1-1，CQ／fj-1和CQ／qj-1。N基因核酸序列和推导氨基酸序列的同源性分析表明所获毒株均归属于基因Ⅰ型狂犬病毒：在基因Ⅰ型的同源性比较中，分离毒株CQ／qj-1与中国狂犬病人用疫苗株3aG型最低，为83．1％，氨基酸水平同源性最低为90．2％（3aG与CQ／ws-1）。分离毒株间低低为97．5％。同源性分析结果还显示出，尽管CQ／WS-1，CQ／wx-1和CQ／fj-1分别来自紧密相邻的3个地区，可是CQ／fj-1却与其它的两个毒株的基因水平同源性最高仅为95．6％。结论 5例临床疑似病例均为犬狂犬病，新分离的动物狂犬病毒流行毒株均为基因Ⅰ型，而且毒株间存在遗传变异。     

The Genetic Characterization of a Novel Natural Recombinant Pseudorabies Virus in China

We sequenced the complete genome of the pseudorabies virus (PRV) FJ epidemic strain, and we studied the characteristics and the differences compared with the classical Chinese strain and that of other countries. Third-generation sequencing and second-generation sequencing technology were used to construct, sequence, and annotate an efficient, accurate PRV library. The complete FJ genome was 143,703 bp, the G+C content was 73.67%, and it encoded a total of 70 genes. The genetic evolution of the complete genome and some key gene sequences of the FJ strain and PRV reference strains were analyzed by the maximum likelihood (ML) method of MEGA 7.0 software. According to the ML tree based on the full-length genome sequences, PRV FJ strain was assigned to the branch of genotype II, and it showed a close evolutionary relationship with PRV epidemic variants isolated in China after 2011. The gB, gC, gD, gH, gL, gM, gN, TK, gI, and PK genes of the FJ strain were assigned to the same branch with other Chinese epidemic mutants; its gG gene was assigned to the same branch with the classic Chinese Fa and Ea strains; and its gE gene was assigned to a relatively independent branch. Potential recombination events were predicted by the RDP4 software, which showed that the predicted recombination sites were between 1694 and 1936 bp, 101,113 and 102,660 bp, and 107,964 and 111,481 bp in the non-coding region. This result broke the previously reported general rule that pseudorabies virus recombination events occur in the gene coding region. The major backbone strain of the recombination event was HLJ8 and the minor backbone strain was Ea. Our results allowed us to track and to grasp the recent molecular epidemiological changes of PRV. They also provide background materials for the development of new PRV vaccines, and they lay a foundation for further study of PRV.     

Use of herpes simplex virus and pseudorabies virus chimeric glycoprotein D molecules to identify regions critical for membrane fusion

Membrane fusion induced by herpes simplex virus (HSV) requires the action of four viral membrane glycoproteins (gB, gD, gH, and gL) and the binding of gD to one of its receptors, such as the herpesvirus entry mediator or nectin-1. The related animal herpesvirus, pseudorabies virus (PRV), encodes a homologous set of glycoproteins and its gD can also use nectin-1 as an entry receptor. We show here that PRV gD, when coexpressed with HSV gB, gH, and gL, cannot substitute for HSV gD in inducing fusion with target cells expressing nectin-1. Chimeric gD molecules composed of HSV and PRV sequences can substitute, provided the first 285 aa are from HSV gD. Because the first 261 aa were sufficient for receptor binding, this suggested that amino acids 262-285 contain a region required for cell fusion but not for receptor binding. Deletions from amino acids 250-299 failed to identify a specific subregion critical for cell fusion, except possibly for amino acids 250-255, which also influenced receptor binding. Instead, presence of a flexible stalk between the membrane and receptor-binding domain appears to be required, perhaps to enable conformational changes in gD on receptor binding and subsequent interactions of undefined regions of gD with the other glycoproteins required for membrane fusion.     

Epidemiological Investigation and Genetic Analysis of Pseudorabies Virus in Yunnan Province of China from 2017 to 2021

In recent years, the prevalence of pseudorabies virus (PRV) has caused huge economic losses to the Chinese pig industry. Meanwhile, PRV infection in humans also sounded the alarm about its cross-species transmission from pigs to humans. To study the regional PRV epidemic, serological and epidemiological investigations of PRV in pig populations from Yunnan Province during 2017–2021 were performed. The results showed that 31.37% (6324/20,158, 95% CI 30.73–32.01) of serum samples were positive for PRV glycoprotein E (gE)-specific antibodies via enzyme-linked immunosorbent assay (ELISA). The risk factors, including the breeding scale and development stage, were significantly associated with PRV seroprevalence among pigs in Yunnan Province. Of the 416 tissue samples collected from PRV-suspected pigs in Yunnan Province, 43 (10.33%, 95% CI 7.41–13.26) samples were positive for PRV-gE nucleic acid in which 15 novel PRV strains from these PRV-positive samples were isolated, whose gC and gE sequences were analyzed. Phylogenetic analysis showed that all 15 isolates obtained in this study belonged to the genotype II. Additionally, the gC gene of one isolate (YuN-YL-2017) was genetically closer to variant PRV strains compared with others, while the gE gene was in the same clade with other classical PRV strains, indicating that this isolate might be a recombinant strain generated from the classical and variant strains. The results revealed the severe PRV epidemic in Yunnan Province and indicated that PRV variants are the major genotypes threatening the pig industry development.     

Whole-Genome Sequence Analysis of Pseudorabies Virus Clinical Isolates from Pigs in China between 2012 and 2017 in China

Pseudorabies virus (PRV) is an economically significant swine infectious agent. A PRV outbreak took place in China in 2011 with novel virulent variants. Although the association of viral genomic variability with pathogenicity is not fully confirmed, the knowledge concerning PRV genomic diversity and evolution is still limited. Here, we sequenced 54 genomes of novel PRV variants isolated in China from 2012 to 2017. Phylogenetic analysis revealed that China strains and US/Europe strains were classified into two separate genotypes. PRV strains isolated from 2012 to 2017 in China are highly related to each other and genetically close to classic China strains such as Ea, Fa, and SC. RDP analysis revealed 23 recombination events within novel PRV variants, indicating that recombination contributes significantly to the viral evolution. The selection pressure analysis indicated that most ORFs were under evolutionary constraint, and 19 amino acid residue sites in 15 ORFs were identified under positive selection. Additionally, 37 unique mutations were identified in 19 ORFs, which distinguish the novel variants from classic strains. Overall, our study suggested that novel PRV variants might evolve from classical PRV strains through point mutation and recombination mechanisms.     

Comparative analyses of the 9 glycoprotein genes found in wild-type and vaccine strains of varicella-zoster virus

The complete DNA sequences of wild-type and vaccine strains of varicella-zoster virus have been published and listed in GenBank. In this comparative genomic analysis, the sequences of the 9 glycoprotein open reading frames (ORFs) were compared. They included gE (ORF68), gI (ORF 67), gC (ORF14), gH (ORF37), gL (ORF60), gB (ORF31), gK (ORF5), gM (ORF50), and gN (ORF8 or ORF9A). After realignment on the basis of newer data, the corrected gB sequence was lengthened to include 931 residues. The data showed that there were glycoprotein polymorphisms that differentiated North American/European strains from Japanese strains-for example, an additional ATG codon in the gL of all Oka strains. Also, there were a small number of coding single-nucleotide polymorphisms present only in glycoproteins of vaccine strains. Because these changes were highly conserved, the structure of the glycoprotein was unlikely to be altered.     

青海省2011-2012年乙型流感病毒HA1基因特性研究

目的 了解2011-2012年青海省乙型流感病毒HA1基因变异情况。方法 随机选择2011-2012年流感病原监测中分离到的乙型流感毒株,提取病毒RNA,通过RT-PCR法扩增病毒HA1基因,纯化产物进行核苷酸序列测定,采用DNA Star 5.0 MegAlign和MEGA4.0生物软件对测序结果进行分析处理,推导出氨基酸序列并进行基因特性分析。结果 12株Victoria系乙流感病毒HA1区域核苷酸序列与2009-2011年WHO推荐疫苗株B/Brisbane/60/2008同源性为88.6%~95.7%,氨基酸替换数在2~5个,所有分离株均在1个相同的氨基酸位点发生了相同的替换,部分分离株在320位减少了一个糖基化位点。5株Yamagata系乙型流感病毒HA1区域核苷酸序列与2011-2012年WHO推荐疫苗株B/Wisconsin/01/2010HA1区相比较同源性为97.7%~99.3%,氨基酸替换数在2-3个,所有分离株均在2个相同的氨基酸位点发生了相同的替换(N116K,D196N),在196位增加了一个糖基化位点。结论 青海省2011-2012年乙型流感病毒HA1基因特性正在逐渐发生变异,但尚未形成抗原漂移,今后在流感监测工作中要加强流感病原学的监测,及时发现新的乙型流感病毒变异株。     

云南省登革4型病毒全基因组序列特征研究

目的阐明云南省2015年5株登革4型病毒(DENV-4)流行株的全基因组序列特征及分子流行病学特点。方法采用C6/36细胞培养法分离病毒,用RT-PCR法扩增新分离DENV-4的全基因组序列,采用ClastalX1.83和MEGA6等生物信息学软件进行核苷酸和推导氨基酸序列同源性及系统进化分析。结果从2015年云南省瑞丽市登革热患者血清中分离到5株DENV-4(本地病例2株,来自缅甸腊戍和南坎市输入性病例3株)。经RT-PCR和序列测定,获得这5株DENV-4的全基因组序列(10 661nt),其开放读码框(103-10 264)编码3 386个氨基酸。全基因组或结构蛋白和非结构蛋白基因序列的系统进化和同源性分析表明,云南分离株间高度同源并聚集为一个进化支,并与泰国不同年代DENV-4基因I型(G-I)流行株具有较近的进化关系和较高的同源性,同属G-I。云南株和泰国株均与同基因型的DENV-4原型株(H241,1956年菲律宾)和中国广州1990年B5株亲缘性和同源性都较低。云南株与H241株在结构蛋白或非结构蛋白中分别存在21和45个氨基酸位点差异。结论首次获得云南省DENV-4分离株的全基因组序列并发现它们与近期东南亚DENV-4G-I流行株亲缘关系较近。首次证实云南省存在DENV-4本地流行,传播来源为相邻缅甸北部边境地区。云南分离株某些氨基酸位点的改变是否与其抗原性和毒力有关尚需进一步研究。     

Differential immunologic reactivity and processing of glycoproteins gA and gB of herpes simplex virus types 1 and 2 made in Vero and HEp-2 cells.

Herpes simplex virus types 1 and 2 (HSV-1 and HSV-2, respectively) specify five major glycoproteins designated as gA, gB, gC, gD, and gE. Previous studies have shown that gA and gB differ in electrophoretic mobility but not in reactivity with antisera prepared to each of these glycoproteins. Moreover, gA and gB of HSV-1 crossreact in serologic tests with the corresponding glycoproteins of HSV-2. In this paper, we report on the reactivities of gA and gB of HSV-1 and HSV-2 with 24 independently derived monoclonal antibodies reactive with these antigens. Our results show the following: (i) Electrophoretic mobilities of HSV-1 and HSV-2 glycoproteins gA and gB made in HEp-2 cells are significantly less than those made in Vero cells. (ii) All monoclonal antibodies precipitated both gA and gB made in infected HEp-2 cells. These include 12 monoclonal antibodies that neutralized virus and 12 that did not. (iii) HSV-2 glycoproteins gA and gB made in HEp-2 cells contain type-specific domains. However, monoclonal antibodies produced by one clone directed to these domains did not react with glycoproteins made in Vero cells. (iv) Lysates of infected Vero cells contain three lower molecular weight polypeptides that also reacted with monoclonal antibodies directed to glycoproteins gA and gB. These polypeptides are virus specific inasmuch as those specified by HSV-1 differ in electrophoretic mobilities from those specified by HSV-2. these polypeptides are absent in lysates of HEp-2 cells.     

中国12株狂犬病街毒株G基因序列测定与分析

目的了解中国狂犬病毒的流行情况以及街毒株与中国人用、兽用狂犬病疫苗株在G基因核苷酸和氨基酸水平的差异,为有效控制狂犬病疫情提供初步科学依据。方法对12株街毒株G基因进行了全基因测序,与其它36株中国狂犬病街毒株,以及中国的疫苗株和其它国家毒株的G基因序列进行了综合分析。结果序列分析表明来源于中国的50株狂犬病毒均为基因Ⅰ型狂犬病毒;其中具有代表性的6株病毒与我国现在使用的各种疫苗株在G基因的核苷酸和氨基酸水平上均存在不同程度的差异,与我国人用疫苗株CTN同源性较高;进化分析表明,我国主要流行狂犬病毒与泰国、印度尼西亚、马来西亚等东南亚狂犬病毒株处于同一分支。中国毒株之间G基因核苷酸同源性分别≥82.3%;氨基酸同源性分别≥92.1%;中国街毒株与疫苗株相比较核苷酸的同源性为79.3%～94.2%,氨基酸的同源性为87.8%～97.9%。结论我国的狂犬病毒为基因Ⅰ型狂犬病毒,可以明确分为6个进化群。无论在核苷酸还是氨基酸水平上,中国多数街毒株与疫苗株CTN之间的同源性要高于与其它疫苗株。     

Chronic-dose acyclovir to suppress frequently recurring genital herpes simplex virus infection: effect on antibody response to herpes simplex virus type 2 proteins

To determine the effect of prolonged suppressive acyclovir therapy on the antibody response to herpes simplex virus type 2 (HSV-2) proteins, we studied sequential sera from 33 patients with frequently recurring (six or more recurrences per year) genital herpes. Twenty-two patients received 400 mg of oral acyclovir and 11 received placebo, twice daily for one year. Sera collected at enrollment, after six months and 12 months of therapy, and during the first recurrence after cessation of therapy were evaluated by western blot for levels of antibodies to HSV-2, gB, gG, gC/gE, VP16, and gD. Mean levels declined by 27%-39% after one year of acyclovir. The magnitude of the decrease in antibody levels was not correlated with disease severity either during or after therapy. Patients with high relative antibody levels to gB after therapy had more-severe first recurrences after therapy than did patients with antibody levels to gB less than or equal to the median. Antibody levels were not restored after the first untreated recurrence.