新疆人类8型疱疹病毒的致病机制研究及在献血者中流行病学调查
发布时间：2019-05-11 17:15
【摘要】：目的:在新疆地区献血员中开展大规模的流行病学横断面调查,掌握HHV-8在该人群中的感染率和危险因素,为评价本地血源质量、制定有针对性的防控措施提供参考数据。进行HHV-8分子病毒学研究,明确基因型和病毒载量特征并建立与KS临床表型的相关性,为指导临床治疗提供科学依据。初步建立本地KS独有的基因差异表达谱并进行功能分析。方法:研究对象分为三类:1)2006年8月至2007年5月收集进入自治区血液中心献血并符合纳入标准的标本4461例;2)1980～2007年本院收集的51例KS组织标本;3)KS病灶和正常皮肤“配对标本”共2对。应用本课题国内首建的HHV-8复合抗原ELISA法筛检献血员人群,统计感染率并应用多元Logistic回归模型预测感染相关危险因素。23例KS组织标本的HHV-8 K1基因经PCR扩增和双向测序后,应用MEGA 3.0和phylip 23.63软件对比GENBANK中同源性高的毒株序列,绘制系统发生树以确定其型别、来源等信息。构建HHV-8 ORF26和内参基因β-actin的重组T载体,应用优化的实时荧光定量PCR法检测HHV-8病毒载量。建立上述指标与KS临床表型的相关性。应用表达芯片检测2对癌与癌旁标本的差异表达基因,为探索新型的诊断或治疗靶点提供资料。结果:HHV-8复合抗原ELISA法其敏感性和特异性分别为81.8%和97.9%,与整病毒免疫荧光法检出率符合度为90.9%。本地献血者中HHV-8平均感染率可达20.4%。民族是最终进入方程的唯一危险因素(p,0.000;OR,1.8;95% CI,1.1-4.2)。HHV-8感染和高效价抗体在民族间的分布差异独立存在。本地HHV-8 K1基因型以A、C为主,且A5、C6和C7为国内首次分离。本地毒株呈现成簇聚集的特征,参考序列主要来自非洲、俄罗斯和中东。C型主要在病程较短的KS中占优势(p,0.046)。HHV-8 ORF26和β-actin起始模板浓度与Ct值之间呈良好的线性关系,相关系数r2=0.996,且重复性较好。不同个体的病毒拷贝数可相差107个数量级。AIDS-KS患者其HHV-8病毒载量显著低于其它两型KS(p,0.025),与抗体效价的分布一致。2对标本共同的表达差异基因为117条,主要与HIV感染、增生、皮肤肿瘤、黑色素瘤、炎症等影响HHV-8感染或KS致瘤过程的已知因素有关。结论:HHV-8复合抗原ELISA法是开展大规模流行病学研究的有力工具。新疆献血员中HHV-8感染率远高于其他省市同类人群,被再次印证属HHV-8流行区,应开展献血前抗HHV-8抗体的筛查。民族背景是导致HHV-8感染和高效价抗体产生的最关键因素。本地HHV-8基因型的类别和分布与少数民族的起源和进化密切相关,且不同型别的致病机制可能不同,具备后期开展遗传易感性研究的基础。成功建立高灵敏性且特异性好的HHV-8实时荧光定量PCR法,可用于确诊临床感染或可疑病例。本地AIDS-KS中病毒载量的高低不能用于预测肿瘤进展,应区别分析该指标在不同类型KS中的作用。表达谱分析显示内质网压力信号通路可能在KS致瘤中起重要作用。
[Abstract]:Objective: to carry out a large-scale epidemiological cross-section survey among blood donors in Xinjiang to master the infection rate and risk factors of HHV-8 in this population, so as to provide reference data for evaluating the quality of local blood sources and formulating targeted prevention and control measures. The molecular virological study of HHV-8 was carried out to clarify the genotypic and viral load characteristics and to establish the correlation with the clinical phenotype of KS, so as to provide scientific basis for guiding clinical treatment. The differential expression profile of genes unique to local KS was established and functional analysis was carried out. Methods: the subjects were divided into three categories: 1) from August 2006 to May 2007, 4461 blood samples were collected from the blood center of the autonomous region and met the inclusion criteria, 2) 51 KS tissue specimens were collected from 1980 to 2007, and 51 samples were collected from our hospital from 1980 to 2007. 3) there were 2 pairs of KS lesions and matched specimens of normal skin. The HHV-8 complex antigen ELISA method, which was first established in China, was used to screen the blood donors. The infection rate was counted and the risk factors related to infection were predicted by multivariate Logistic regression model. The HHV-8 K1 gene in 23 KS tissue specimens was amplified by PCR and sequenced bidirectionally. MEGA 3.0 and phylip 23.63 software were used to compare the sequences of strains with high homology in GENBANK, and the phylogenetic tree was drawn to determine the type and source of the strains. The recombinant T vector of HHV-8 ORF26 and internal reference gene 尾-actin was constructed and the load of HHV-8 virus was detected by optimized real-time fluorescence quantitative PCR. To establish the correlation between the above indexes and the clinical phenotype of KS. Two pairs of differentially expressed genes between cancer and paracancerous specimens were detected by expression chip, which provided data for exploring new diagnostic or therapeutic targets. Results: the sensitivity and specificity of HHV-8 complex antigen ELISA method were 81.8% and 97.9%, respectively, and the coincidence rate with the detection rate of whole virus immunofluorescence was 90.9%. The average infection rate of HHV-8 in local blood donors was 20.4%. Nationality is the only risk factor for entering the equation (p, 0.000 OR, 1.8% CI,1.1-4.2). There are independent differences in the distribution of HHV-8 infection and high valence antibodies among ethnic groups. The local HHV-8 K1 genotype was mainly A, C, and A5, C 6 and C 7 were isolated for the first time in China. The local strains showed the characteristics of cluster aggregation, and the reference sequences were mainly from Africa, Russia and the Middle East. C was mainly dominant in KS with a short course of disease (p, 0.046). There was a good linear relationship between the initial template concentration of HHV-8 ORF26 and 尾-actin and KT value, and the correlation coefficient R2 was 0.996, and the reproducibility was good. The viral copy number of different individuals varied by 107 orders of magnitude. The HHV-8 viral load of AIDS-KS patients was significantly lower than that of the other two types of KS, which was consistent with the distribution of antibody titer. 2 the common expression differentially expressed genes were 117, It is mainly related to HIV infection, proliferation, skin tumor, melanoma, inflammation and other known factors affecting the tumorigenic process of HHV-8 infection or KS. Conclusion: HHV-8 complex antigen Elisa is a powerful tool for large-scale epidemiological research. The infection rate of HHV-8 in Xinjiang blood donors is much higher than that in other provinces and cities. It has been confirmed that it belongs to HHV-8 epidemic area again. The screening of anti-HHV-8 antibody before blood donation should be carried out. Ethnic background is the most key factor leading to HHV-8 infection and high valence antibody production. The types and distribution of local HHV-8 genotypes are closely related to the origin and evolution of ethnic minorities, and the pathogenic mechanisms of different types may be different, which has the basis of genetic susceptibility research in the later stage. A highly sensitive and specific HHV-8 real-time fluorescence quantitative PCR method was successfully established, which can be used to diagnose clinical infection or suspicious cases. The level of viral load in local AIDS-KS could not be used to predict the progression of tumor. The role of the index in different types of KS should be analyzed differentially. Expression profile analysis showed that endoplasmic reticulum pressure signaling pathway may play an important role in the tumorigenicity of KS.
【学位授予单位】：新疆医科大学
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R181.3
本文编号：2474692
[Abstract]:Objective: to carry out a large-scale epidemiological cross-section survey among blood donors in Xinjiang to master the infection rate and risk factors of HHV-8 in this population, so as to provide reference data for evaluating the quality of local blood sources and formulating targeted prevention and control measures. The molecular virological study of HHV-8 was carried out to clarify the genotypic and viral load characteristics and to establish the correlation with the clinical phenotype of KS, so as to provide scientific basis for guiding clinical treatment. The differential expression profile of genes unique to local KS was established and functional analysis was carried out. Methods: the subjects were divided into three categories: 1) from August 2006 to May 2007, 4461 blood samples were collected from the blood center of the autonomous region and met the inclusion criteria, 2) 51 KS tissue specimens were collected from 1980 to 2007, and 51 samples were collected from our hospital from 1980 to 2007. 3) there were 2 pairs of KS lesions and matched specimens of normal skin. The HHV-8 complex antigen ELISA method, which was first established in China, was used to screen the blood donors. The infection rate was counted and the risk factors related to infection were predicted by multivariate Logistic regression model. The HHV-8 K1 gene in 23 KS tissue specimens was amplified by PCR and sequenced bidirectionally. MEGA 3.0 and phylip 23.63 software were used to compare the sequences of strains with high homology in GENBANK, and the phylogenetic tree was drawn to determine the type and source of the strains. The recombinant T vector of HHV-8 ORF26 and internal reference gene 尾-actin was constructed and the load of HHV-8 virus was detected by optimized real-time fluorescence quantitative PCR. To establish the correlation between the above indexes and the clinical phenotype of KS. Two pairs of differentially expressed genes between cancer and paracancerous specimens were detected by expression chip, which provided data for exploring new diagnostic or therapeutic targets. Results: the sensitivity and specificity of HHV-8 complex antigen ELISA method were 81.8% and 97.9%, respectively, and the coincidence rate with the detection rate of whole virus immunofluorescence was 90.9%. The average infection rate of HHV-8 in local blood donors was 20.4%. Nationality is the only risk factor for entering the equation (p, 0.000 OR, 1.8% CI,1.1-4.2). There are independent differences in the distribution of HHV-8 infection and high valence antibodies among ethnic groups. The local HHV-8 K1 genotype was mainly A, C, and A5, C 6 and C 7 were isolated for the first time in China. The local strains showed the characteristics of cluster aggregation, and the reference sequences were mainly from Africa, Russia and the Middle East. C was mainly dominant in KS with a short course of disease (p, 0.046). There was a good linear relationship between the initial template concentration of HHV-8 ORF26 and 尾-actin and KT value, and the correlation coefficient R2 was 0.996, and the reproducibility was good. The viral copy number of different individuals varied by 107 orders of magnitude. The HHV-8 viral load of AIDS-KS patients was significantly lower than that of the other two types of KS, which was consistent with the distribution of antibody titer. 2 the common expression differentially expressed genes were 117, It is mainly related to HIV infection, proliferation, skin tumor, melanoma, inflammation and other known factors affecting the tumorigenic process of HHV-8 infection or KS. Conclusion: HHV-8 complex antigen Elisa is a powerful tool for large-scale epidemiological research. The infection rate of HHV-8 in Xinjiang blood donors is much higher than that in other provinces and cities. It has been confirmed that it belongs to HHV-8 epidemic area again. The screening of anti-HHV-8 antibody before blood donation should be carried out. Ethnic background is the most key factor leading to HHV-8 infection and high valence antibody production. The types and distribution of local HHV-8 genotypes are closely related to the origin and evolution of ethnic minorities, and the pathogenic mechanisms of different types may be different, which has the basis of genetic susceptibility research in the later stage. A highly sensitive and specific HHV-8 real-time fluorescence quantitative PCR method was successfully established, which can be used to diagnose clinical infection or suspicious cases. The level of viral load in local AIDS-KS could not be used to predict the progression of tumor. The role of the index in different types of KS should be analyzed differentially. Expression profile analysis showed that endoplasmic reticulum pressure signaling pathway may play an important role in the tumorigenicity of KS.
【学位授予单位】：新疆医科大学
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R181.3
【引证文献】
相关期刊论文 前1条
1 冷弘;王媛媛;燕备战;方春霞;马晶;臧文巧;李敏;赵国强;;洛阳地区无偿献血人群中疱疹病毒8型的检测[J];广东医学;2012年20期
，本文编号：2474692
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