1型单纯疱疹病毒(Herpes simplex virus ，HSV1)是疱疹病毒科α病毒亚科的一员，能引起多种疾病，包括龈口炎，病毒性脑炎，角膜结膜炎等，能够感染多种细胞并在体内感染中建立潜伏感染。针对HSV感染目前还没有疫苗和完全清除体内病毒的药物被研发出来。此外，HSV由于其具有大型环状DNA和神经潜伏感染的特征，在研究神经系统疾病时可以作为载体而得到广泛的应用。

本研究中从临床病人口唇部疱液分离得到一株病毒，通过Vero细胞扩增纯化，通过PCR分析、电镜鉴定确定该病毒为1型HSV，命名为HSV1-ZW6，对其进行病毒增值特性分析，包括对其感染不同细胞病变形态学分析、一步生长曲线的绘制、小鼠神经毒力检测试验等，结果提示此分离株与实验室传代毒株HSV1-8F相比，其神经毒力略高于HSV1-8F。利用高通量测序技术对此株病毒基因组进行了测序，联合Sanger测序对其进行修补，最终获得了150290bp的病毒全基因组序列，并通过对全基因进行基因预测和注释获得其全基因组结构图，选取部分基因进行亲缘关系进化树绘制和简单重复序列分析，发现位于IRL/TRL区的一段32个A/T重复的特征序列。根据已知序列，利用CRISPR-Cas9技术快速获得了一株带有绿色荧光蛋白的重组病毒ZW6-EGFP。此外，本研究在建立了二倍体细胞KMB17细胞代次与端粒长度变化模型的基础上，在细胞水平检测了单纯疱疹病毒感染KMB17细胞后，对感染后端粒长度的影响，初步发现HSV1感染KMB17细胞后会对其产生影响，而感染癌细胞或永生化细胞系则未发现相似结果。

通过对HSV1的分离、鉴定及全基因组测序完成，本研究建立了一株新的HSV1参考株及其相应的报告病毒，利用该病毒研究病毒与细胞之间的相互作用进行了初步探索，更多的生物特性还有待进一步的深入研究。

Herpes simplex virus (HSV) type 1 is a member of the herpesviridae family，the virus frequently infects human beings,causing a range of diseases,including gingival stomatitis;encephalitis;corneal conjunctivitis. HSV1 can productively infect many different types of cells and establish latent infections in neuronal ganglia in vivo.There is no vaccine against HSV infection and complete removal of the virus drug is developed. In addition,HSV is characterized by large cyclic DNA and latent infection,the HSV-vectors are especially useful for delivery to the nervous system.

    In this study ,a strain of virus was isolated from blisters of patient use Vero,and the isolated virus is proved to be HSV-1 through Sequence analysis and Electron microscopy identification,named HSV1-ZW6 strain.For proliferative rule of HSV1-ZW6,including the infection of different cell line,one step growth curve drawing,mouse neurovirulence test etc.The results show that the neurovirulence of this isolated strain higher than the passage strain HSV1-8F.The genome of this virus are completed by high-throughput sequencing technologise and repairing with Sanger method,for gene prediction and annotation to obtain the structure graph of genome ,completing phylogenetic tree  based on parts of gene ,then found a special 32bp repeat sequence in IRL/TRL based on analysis of sinple sequence  repeats. According to the known sequence,a recombinant virus ZW6-EGFP was successfully obtained via CRISPR-Cas9 system. In addition, to studing KMB17 cells generation and changes in telomere length, a diploid cell model is established .Then,at the cellular level to detect the KMB17 cells infect with herpes simplex virus , preliminary findings HSV1 infect KMB17 can effect the telemore length, and cancer cells or immortalized cell lines is not found similar results.

Through the isolation,identification and complete genome sequencing of HSV1, this study establishes a strain of HSV1 reference strain and its report virus .We just have a preliminary exploration of HSV1-ZW6,there are many worthy of study above this virus. The study is only a preliminary exploration on the interaction between virus and cell, the reference  strains of biological characteristics  needs to be more studied further