济南地区无偿献血者中血源性传播病毒感染的流行病学调查及人疱疹病毒8型原核表达载体的构建

发布时间：2018-05-24 03:47

  本文选题：献血者 + HIV感染　； 参考：《山东大学》2008年硕士论文

【摘要】： 第一部分济南地区无偿献血者中血源性传播病毒感染的流行病学调查 目的:检测济南地区无偿献血者血浆中乙型肝炎病毒表面抗原(HBsAg)、抗丙型肝炎病毒(HCV)抗体、抗人类免疫缺陷病毒(HIV)抗体以及抗人疱疹病毒8型(HHV-8)IgG抗体,分析我国济南地区献血人群中相关病毒的感染情况和流行病学特点;为济南地区无偿献血人群的选择和制定招募安全血源的策略提供理论依据。 方法: 1.血清学检测 1.1利用ELISA方法对2006年1月～2006年12月无偿献血者的血浆标本(43904份)进行HBsAg、抗-HCV、抗-HIV的检测;抗-HIV检测呈阳性者送山东省疾病预防控制中心进行Western-Blot确认。 1.2利用ELISA方法对其中520份献血者血浆进行抗-HHV-8 IgG抗体的检测,包被抗原为HHV-8结构蛋白ORF K8.1合成多肽。 2.统计学处理 2006年1月～2006年12月的献血者基本资料和各项试验检测的结果应用SPSS10.0统计软件包进行统计学分析。统计学方法采用x~2检验。 结果: 1.43904份献血者血浆中HBsAg检出率为2.04%(898/43904),抗-HCV检出率为0.78%(344/43904),抗-HIV检出率为0.0045%(2/43904)。520份献血者血浆中抗-HHV-8检出率为5.962%(31/520)。 2.统计学分析结果显示,HBsAg、抗-HCV检出率在不同年龄组间的差异无统计学意义(P＞0.05),在不同职业间的差异有统计学意义(P＜0.05);男性献血者中HBsAg的检出率高于女性,差异有统计学意义(P＜0.05);不同性别、年龄献血者血浆中抗-HHV-8的检出率差异无统计学意义(P＞0.05)。 结论:济南市无偿献血者中存在一定比率的HCV感染者及相对较高的HBV、HHV-8感染者,同时尚存在个别HIV感染者。在以后的血液筛查工作中有必要进一步提高检测方法的准确度以及加强招募前的筛查工作,以不断提高输血安全。 第二部分人疱疹病毒8型ORF73C基因原核表达载体的构建 目的: 构建人疱疹病毒8型ORF73C基因原核表达载体,为下一步的基因表达及重组蛋白诊断试剂盒的制备奠定基础。 方法: 根据HHV-8 ORF73C基因序列和相关文献设计PCR引物,在引物的5'端分别引入限制酶Bam HI和HindⅢ酶切位点,以含有HHV-8 ORF73基因片段的重组质粒pcDNAHis.LANA1为模板,利用PCR方法扩增HHV-8 ORF73C基因,克隆至pGEM T-easy载体,并进一步克隆至原核表达载体pET30(a+)中,构建原核表达质粒pETORF73C。 结果: 1.琼脂糖凝胶电泳证实PCR扩增产物为目的基因(HHV-8 ORF73C)。 2.DNA测序证实HHV-8 ORF73C原核表达载体pETORF73C构建成功。 结论本研究成功构建了含有HHV-8 ORF73C抗原表位的原核表达质粒pETORF73C,为以后的重组蛋白制备奠定了基础。
[Abstract]:Part I Epidemiological investigation of blood-borne sexually transmitted virus infection among unpaid blood donors in Jinan Objective: to detect hepatitis B virus surface antigen (HBsAg), anti-hepatitis C virus (HCV) antibody, anti-human immunodeficiency virus (HIV) antibody and anti-human herpesvirus 8 (HHV-8) IgG in plasma of blood donors in Jinan area. To analyze the infection and epidemiological characteristics of related viruses in blood donors in Jinan, and to provide a theoretical basis for the selection of free blood donors in Jinan and the formulation of strategies for recruiting safe blood sources. Methods: 1. Serological detection 1.1 A total of 43904 plasma samples from unpaid blood donors from January 2006 to December 2006 were tested for HBsAg, anti-HCV and anti-HIV by ELISA method, and those who were positive for anti-HIV were sent to Shandong Center for Disease Prevention and Control for Western-Blot confirmation. 1.2 the anti-HHV-8 IgG antibody was detected in 520 blood donor plasma by ELISA method. The antigen was HHV-8 structural protein ORF K8.1 synthetic polypeptide. 2. Statistical processing From January 2006 to December 2006, the basic data of blood donors and the results of various tests were statistically analyzed by SPSS10.0 statistical software package. The statistical method was x2 test. Results: The detection rate of HBsAg in plasma of 1.43904 blood donors was 2.04. The positive rate of anti-HCV was 0.78. The detection rate of anti-HCV was 344 / 43904. The detection rate of anti-HIV was 0.0045%. The detection rate of anti-HHV-8 was 5.962%. 2. The results of statistical analysis showed that there was no significant difference in detection rate of HBsAg and anti-HCV among different age groups (P > 0.05), but there was significant difference among different occupations (P < 0.05), and the detection rate of HBsAg in male blood donors was higher than that in women. The difference was statistically significant (P < 0.05), but there was no significant difference in the detection rate of anti-HHV-8 in blood donors of different sex and age (P > 0.05). Conclusion: there are a certain percentage of HCV infection and a relatively high rate of HBV HHV-8 infection among unpaid blood donors in Jinan City, and there are still individual HIV infected individuals. In order to improve the safety of blood transfusion, it is necessary to further improve the accuracy of detection methods and the screening work before recruitment in the future blood screening work. Construction of prokaryotic expression vector of human herpesvirus type 8 ORF73C gene Objective: The prokaryotic expression vector of human herpesvirus type 8 ORF73C gene was constructed, which laid a foundation for the next step of gene expression and preparation of recombinant protein diagnostic kit. Methods: According to the sequence of HHV-8 ORF73C gene and related literatures, PCR primers were designed. Restriction enzyme Bam HI and Hind 鈪，

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