本文關鍵詞：異基因造血干細胞移植人皰疹病毒7型感染的研究　出處：《北京中醫(yī)藥大學》2011年碩士論文　論文類型：學位論文

  更多相關文章： 實時熒光定量PCR 人類皰疹病毒7型 異基因造血干細胞移植 感染 病毒載量

【摘要】：人皰疹病毒7型(HHV-7),屬于β-皰疹病毒家族(DNA病毒)。該病毒具有潛伏感染的特點,世界各地的流行病學調查顯示,不同地區(qū)人群血液和唾液中陽性率達17%～96%,提示HHV-7是一種普遍存在的病毒。HHV-7感染與幼兒急疹、中樞神經系統(tǒng)感染、器官移植、慢性EB病毒樣感染、疲勞綜合征和獲得性免疫缺陷綜合征患者的合并感染、扁平苔蘚相關。其原發(fā)感染與幼兒急疹有關,原發(fā)感染后病毒可在體內長期潛伏,器官移植后患者HHV-7可能出現再活化。本項研究旨在建立實時熒光定量PCR(RT-PCR)檢測人類皰疹病毒7型(HHV-7)方法并評價其敏感性和特異性,從而定量測量臨床標本,對異基因造血干細胞移植((allo-HSCT)感染情況進行研究。 1實時熒光定量PCR檢測人類皰疹病毒7型方法的建立 目的建立實時定量PCR檢測人類皰疹病毒7型的方法,用于臨床HHV-7的基因定量檢測。方法依據HHV-7基因序列設計引物和探針,擴增HHV-7特異的基因片段,克隆重組到pGM-T載體,基因序列測定,重組質粒作為標準品,建立標準曲線,進行靈敏度和特異性的檢測。結果PCR擴增獲得128bp片段,序列測定重組質粒含有特異的HHV-7基因序列,標準曲線的相關系數為0.9989,實時定量PCR擴增效率為95.86%,檢測靈敏度為20 copies/反應體系。結論成功建立了HHV-7基因實時定量PCR檢測的方法,具有高度敏感性和特異性,可用于臨床和科研標本HHV-7的基因定量檢測。 2異基因造血干細胞移植人皰疹病毒7型感染的研究 目的探索HHV-7在異基因造血干細胞移植患者的感染現狀。方法采集40例allo-HSCT移植前和移植后患者外周血標本384份以及40份供者標本。采用實時熒光定量PCR檢測供者以及移植前和移植后患者人類皰疹病毒7型。結果40位allo-HSCT供者DNA陽性率為32.5%(13/40),病毒載量中位數為271EqCop/106PBC(37～1095)；40位allo-HSCT受者,移植前DNA陽性率為37.5%(15/40),病毒載量中位數為285EqCop/106PBC(47～3764)；40位allo-HSCT受者,344份標本中,移植后DNA標本陽性率43.90%(151/344),病毒載量中位數為457EqCop/106PBC(41～5218),顯著高于供者病毒載量(p0.05)。HHV-7 DNA陽性率與性別、年齡、基礎疾病等無顯著相關性。受者移植后HHV-7感染與供者、受者移植前感染顯著相關(p0.05)。結論HHV-7感染在allo-HSCT后患者中很常見,其病毒載量高于供者。供者、受者移植前HHV-7陽性者,受者移植后HHV-7更容易發(fā)生感染,因而供者、受者移植前HHV-7陽性者,移植后患者需要檢測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.

【學位授予單位】：北京中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R373

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