异基因造血干细胞移植人疱疹病毒7型感染的研究

发布时间：2018-01-01 14:02

本文关键词：异基因造血干细胞移植人疱疹病毒7型感染的研究　出处：《北京中医药大学》2011年硕士论文　论文类型：学位论文

【摘要】：人疱疹病毒7型(HHV-7),属于β-疱疹病毒家族(DNA病毒)。该病毒具有潜伏感染的特点,世界各地的流行病学调查显示,不同地区人群血液和唾液中阳性率达17%～96%,提示HHV-7是一种普遍存在的病毒。HHV-7感染与幼儿急疹、中枢神经系统感染、器官移植、慢性EB病毒样感染、疲劳综合征和获得性免疫缺陷综合征患者的合并感染、扁平苔藓相关。其原发感染与幼儿急疹有关,原发感染后病毒可在体内长期潜伏,器官移植后患者HHV-7可能出现再活化。本项研究旨在建立实时荧光定量PCR(RT-PCR)检测人类疱疹病毒7型(HHV-7)方法并评价其敏感性和特异性,从而定量测量临床标本,对异基因造血干细胞移植((allo-HSCT)感染情况进行研究。 1实时荧光定量PCR检测人类疱疹病毒7型方法的建立目的建立实时定量PCR检测人类疱疹病毒7型的方法,用于临床HHV-7的基因定量检测。方法依据HHV-7基因序列设计引物和探针,扩增HHV-7特异的基因片段,克隆重组到pGM-T载体,基因序列测定,重组质粒作为标准品,建立标准曲线,进行灵敏度和特异性的检测。结果PCR扩增获得128bp片段,序列测定重组质粒含有特异的HHV-7基因序列,标准曲线的相关系数为0.9989,实时定量PCR扩增效率为95.86%,检测灵敏度为20 copies/反应体系。结论成功建立了HHV-7基因实时定量PCR检测的方法,具有高度敏感性和特异性,可用于临床和科研标本HHV-7的基因定量检测。 2异基因造血干细胞移植人疱疹病毒7型感染的研究目的探索HHV-7在异基因造血干细胞移植患者的感染现状。方法采集40例allo-HSCT移植前和移植后患者外周血标本384份以及40份供者标本。采用实时荧光定量PCR检测供者以及移植前和移植后患者人类疱疹病毒7型。结果40位allo-HSCT供者DNA阳性率为32.5%(13/40),病毒载量中位数为271EqCop/106PBC(37～1095)；40位allo-HSCT受者,移植前DNA阳性率为37.5%(15/40),病毒载量中位数为285EqCop/106PBC(47～3764)；40位allo-HSCT受者,344份标本中,移植后DNA标本阳性率43.90%(151/344),病毒载量中位数为457EqCop/106PBC(41～5218),显著高于供者病毒载量(p0.05)。HHV-7 DNA阳性率与性别、年龄、基础疾病等无显著相关性。受者移植后HHV-7感染与供者、受者移植前感染显著相关(p0.05)。结论HHV-7感染在allo-HSCT后患者中很常见,其病毒载量高于供者。供者、受者移植前HHV-7阳性者,受者移植后HHV-7更容易发生感染,因而供者、受者移植前HHV-7阳性者,移植后患者需要检测HHV-7。
[Abstract]:Herpes simplex virus type 7 (HHV-7), belonging to the beta herpesvirus (DNA virus). The virus has the characteristics of latent infection, epidemiology shows around the world, the positive rate of blood and saliva of different regions ranged from 17% to 96%, suggesting that HHV-7 is a ubiquitous virus.HHV-7 infection and exanthema subitum the central nervous system, infection, organ transplantation, chronic EB virus infection, fatigue syndrome and acquired immunodeficiency syndrome complicated with combined infection in patients with lichen planus. The primary infection with exanthema subitum after primary infection, the virus can lurk in the body, organ transplant patients after HHV-7 activation. This may occur again the study aims to establish a real-time fluorescent quantitative PCR (RT-PCR) detection of human herpes virus type 7 (HHV-7) method and to evaluate its sensitivity and specificity, and quantitative measurement of clinical specimens of allogeneic hematopoietic stem cell transplantation ((all O-HSCT) the infection was studied.
The establishment of 1 human herpes virus type 7 method by real time fluorescence quantitative PCR
Objective to establish a method for detection of human herpes virus type 7 real-time quantitative PCR, HHV-7 gene for quantitative clinical detection. Methods according to the sequence of HHV-7 gene primers and probes were designed to amplified HHV-7 gene fragments specific, cloned and recombined into pGM-T vector, gene sequencing, the recombinant plasmid as a standard, to establish the standard curve, detection sensitivity and the specificity of the results. The PCR amplified 128bp fragment, HHV-7 gene sequence determination of recombinant plasmids containing specific, the correlation coefficient of the standard curve is 0.9989, real time quantitative PCR amplification efficiency was 95.86%, the detection sensitivity of 20 copies/ reaction system. Objective to establish the method of real-time quantitative PCR detection of HHV-7 gene, with high sensitivity and specific, can be used for quantitative gene research and clinical specimens of HHV-7 detection.
Study on human herpes virus type 7 infection by 2 allogeneic hematopoietic stem cell transplantation
Objective to investigate the infection status of cell transplantation in allogeneic hematopoietic stem HHV-7. Methods to collect 40 cases of allo-HSCT before transplantation and after transplantation in patients with peripheral blood specimens of 384 copies and 40 copies of donor specimens. Using real-time fluorescence quantitative PCR detection of donors and recipients before and after transplantation with human herpesvirus 7. Results 40 allo-HSCT for the positive rate of DNA was 32.5% (13/40), viral load was 271EqCop/106PBC (37 ~ 1095); 40 allo-HSCT recipients, the positive rate of DNA before transplantation was 37.5% (15/40), viral load was 285EqCop/106PBC (47 ~ 3764); 40 allo-HSCT recipients, 344 specimens, transplantation after the positive rate of DNA were 43.90% (151/344), viral load was 457EqCop/106PBC (41 ~ 5218), significantly higher than that of donor viral load (P0.05).HHV-7 DNA positive rate and the gender, age, basic diseases such as no correlation. HHV-7 infection after transplantation. With the donor, recipients before transplantation infection significantly correlated (P0.05). Conclusion HHV-7 infection is common in allo-HSCT patients, the viral load is higher than that of the donor. The donor and recipient before transplantation HHV-7 positive recipients after transplantation, HHV-7 is more prone to infection, so the donor and recipient before transplantation HHV-7 positive, post transplant patients need to detect HHV-7.

【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R373

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