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1.
目的:探讨异基因造血干细胞移植(allo—HSCT)后巨细胞病毒感染的早期诊断及人类巨细胞病毒血症(Cy—tomegalovirus antigenemla)与巨细胞病毒病(CMVdisease)发生的关系。方法;CMVpp65单克隆抗体荧光免疫组织化学法检测CMV早期抗原,并进行动态观察。结果:47例I{SCT中,17例(36.17%)发生巨细胞病毒血症,7例(14.89%)发生CMV病。基础疾病为血液系统恶性疾病的患者,进行异基因造血干细胞移植后,巨细胞病毒抗原血症和巨细胞病毒病的发生率分别为54.55%和31.82%,而血液系统非恶性疾病的患者分别为20.00%和0.00%,两者之间差异有显著性(P〈0.05)。基础疾病、急性GVHD是巨细胞病毒抗原血症和巨细胞病毒病发生的危险因素。结论:CMVpp65病毒抗原检测在造血干细胞移植后CMV病的早期诊断、早期预防、早期治疗及延长移植后长期生存率具有重要意义。  相似文献   

2.
目的:探讨同种异基因造血干细胞移植(allo-HSCT)中巨细胞病毒(cytomegalovirus,CMV)激活情况及激活相关因素。方法:52例接受allo-HSCT的患者为研究对象,所有患者及其供者移植前均为CMV携带者,即为CMV血清型阳性。所有移植术后患者应用实时定量聚合酶链反应(PCR)方法对CMVDNA进行定量检测。结果:移植后100d内,52例患者中28例检测到CMVDNA复制。造血干细胞移植(HSCT)供者类型和移植物抗宿主反应(graft versus host disease,GVHD)的发生与CMV激活明显相关(P值分别为0.018和0.039)。在44例可评估患者中,allo-HSCT100d后仅7例检测到CMVDNA复制,其发生率与移植后发生GVHD且接受激素治疗(P=0.0009)、人类白细胞抗原(HLA)全相合无关供者移植或者HLA部分相合同胞供者移植有关(P=0.037)。应用实时定量PCR检测发现,CMV对更昔洛韦抢先治疗的应答存在2种动力学模式(快速型和缓慢型),其中表现为缓慢型的患者CMV最高负载量较快速型高10倍以上,其病毒衰减时间也要延长3~4周,多数发生在因出现GVHD而接受激素治疗以及接受HLA全相合无关供者和HLA部分相合同胞供者干细胞移植的患者。结论:移植前CMV血清型阳性患者接受allo-HSCT后100d内出现CMV激活是常见并发症。移植100d之后,对于未发生GVHD而未使用激素治疗者或以HLA全相合同胞为供体的患者,CMV激活概率低,无需进行常规频繁监测。  相似文献   

3.
本研究比较巨细胞病毒(CMV)定量PCR检测和CMV—pp65抗原检测在异基因造血干细胞移植CMV感染中的诊断价值。以84例异基因造血干细胞移植患者为研究对象,自预处理开始每周对患者EDTA抗凝外周血血标本进行CMV—pp65抗原及cMV定量PCR动态检测,直至出院,比较观察两者在诊断CMV感染中的作用。结果表明:84例移植患者的732份系列血标本中,26例移植后检出CMV定量PCR阳性,检出率30.95%,其中9例为cMV血症,13例为cMV病,检出中位时间为37.1(7—105)天;22例CMVpp65抗原检测阳性,检出率26.19%,检出中位时间为46.6(10—128)天;所有CMV—pp65抗原检测阳性患者CMV定量PCR均为阳性,4例CMV定量PCR阳性但CMV—pp65抗原检测阴性患者均未发展为CMV病。CMV病多出现于CMV定量PCR中等至高拷贝数或中等至高水平病毒血症(CMV—pp65抗原)病例中。治疗后CMV定量PCR转阴中位时间为17.5(11—28)天。CMV—pp65抗原转阴中位时间为10.0(7—21)天。结论:CMV定量PCR和CMV—pv65抗原检测均能作为异基因造血干细胞移植CMV感染早期诊断的有效手段,CMV定量PCR更敏感,CMV—pp65抗原检测更特异,两者同时使用能更有效地提高CMV感染诊断水平,监控其发生发展。  相似文献   

4.
本研究探讨异基因造血干细胞移植(allo HSCT)后受者外周血白细胞巨细胞病毒(CMV)pp65抗原阳性细胞数的变化及临床意义。以104例HSCT受者为研究对象,采集移植前后及恢复期受者的抗凝血标本,用CMVpp65单克隆抗体荧光免疫组织化学法检测CMV早期抗原,并进行动态观察。结果表明:104例HSCT受者中29例移植后检出CMV pp65抗原阳性,检出率27.88%,其中16例为CMV血症,13例为CMV病。29例经抗病毒治疗后25例抗原转为阴性,有效率86.21%,死亡4例,病死率13.79%。CMV感染患者中,受非血缘或单倍体相合异基因HSCT的患者与受同胞全相合异基因HSCT的患者CMV感染率分别为39.29%和14.58%,两者相比较有显著性差异(P〈0.05)。0-Ⅰ度急性GVHD组CMV感染的发生率为19.44%,而Ⅱ-Ⅳ急性GVHD组为46.88%,两者间的差异有显著意义(P〈0.05)。CMV抗原阳性与阴性患者的急性GVHD发生率有显著性差异(P〈0.05)。结论:外周血白细胞CMV pp65病毒抗原检测可作为造血干细胞移植后CMV病的监测指标,用于指导移植后抗病毒的治疗和疗效评价。  相似文献   

5.
本研究探讨同种异基因造血干细胞移植(allo-HSCT)后早期有效检测巨细胞病毒(CMV)感染的方法。应用荧光定量PCR和ELISA试剂盒分别检测19名allo-HSCT受者,214份标本的血浆DNA负荷量和血清IgM抗体,同时应用流式细胞术检测188份标本白细胞pp65抗原。结果表明:pp65抗原、DNA定量和IgM抗体的阳性检出率分别为30.85%(58/188)、35.51%(76/214)和13.08%(28/214),连续阳性病例和临床诊断的符合率分别为7/8、7/8和3/8。DNA定量与pp65抗原阳性检出率的差别无统计学意义(P〉0.05),但两种检测方法有明显的相关性(P〈0.05)。IgM抗体阳性检出率明显低于DNA定量和pp65抗原,其差别均有统计学意义(P〈0.05),与另两种检测方法虽有关系,但不密切。结论:流式细胞术和荧光定量PCR检测allo-HSCT受者CMV早期感染可靠、简便快速,值得临床推广使用。  相似文献   

6.
背景:异基因造血干细胞移植后EB病毒再激活可导致致死性的移植后淋巴细胞增殖性疾病及相关疾病,目前国内尚未建立完整的EB病毒再激活及其相关性疾病的诊疗体系。目的:前瞻性研究异基因造血干细胞移植后患者EB病毒再激活的发生和相关危险因素。方法:纳入129例接受异基因造血干细胞移植的患者,采用实时定量PCR方法定期测定外周血中EB病毒载量,Kaplan-Meier模型分析其再激活的发生率,Logistic回归分析模型分析其再激活的相关危险因素。结果与结论:异基因造血干细胞移植后EB病毒再激活及其相关疾病发生率高,EB病毒再激活发生的危险因素有HLA配型不合、应用抗胸腺细胞球蛋白、Ⅲ~Ⅳ度急性移植物抗宿主病及年龄小于20岁。  相似文献   

7.
为了解人类疱疹病毒6型(human herpesvirus 6,HHV-6)在中国接受异基因造血干细胞移植(hematopoietic stem cell transplantation,HSCT)的人群中感染现状及其与巨细胞病毒(cytomegalovirus,CMV)感染的相关性,对72例接受HSCT的患者HHV-6DNA血症进行连续监测。收集HSCT患者移植前和移植后1-12周EDTA抗凝的外周血标本共680份,采用巢式聚合酶链反应检测外周血单个核细胞中HHV-6DNA,并利用Hind Ⅲ限制性内切酶对HHV-6进行基因分型,同时采用免疫荧光法检测CMV抗原血症。结果显示,HSCT后62.5%(45/72)的患者至少1次出现HHV-6DNA血症,首次检出的中位时间为14(7-63)天;除1例患者检出HHV-6A型以外,其余所有HHV-6阳性患者均为HHV-6B型感染。65.3%(47/72)的移植后患者至少1次发生CMV抗原血症,首次检出的中位时间为43(14-105)天。HHV-6与CMV共感染(CMV^+/HHV-6^+)的发生率为52.8%(38/72).HHV-6DNA血症的首次检出时间早于CMV抗原血症(P〈0.0001)。HHV-6DNA血症阳性患者CMV抗原血症检出率显著高于HHV-6血症阴性患者[84.4%(38/45)vs33.3%(9/27),P=0.0001]。HSCT后的疱疹病毒感染相关疾病中出血性膀胱炎(HC)发生率较高[23.6%(17/72)],其中88.2%(15/17)的HC发生于HHV-6血症阳性患者.82.3%(14/17)发生于CMV^+/HHV-6^+患者。结论:HSCT后HHV-6感染以及HHV-6与CMV共感染状态普遍存在,且发生于移植后早期的HHV-6感染与发生时间相对较晚的CMV感染之间可能存在相关性。  相似文献   

8.
Real-time polymerase chain reaction (PCR) and pp65 antigenemia assay for the detection of active cytomegalovirus infection in immunocompromised patients experiencing neutropenia after bone marrow or kidney transplantation have been compared with a special focus on evaluability and embedment in daily routine diagnostics. Investigating 334 specimens from 97 patients, real-time PCR was shown to be the superior assay with regard to the parameters focused on.  相似文献   

9.
Development of highly sensitive quantitative assays for cytomegalovirus (CMV) DNA detection is crucial for identification of immunodeficient patients at high risk of CMV disease. We designed 2 internally controlled competitive quantitative assays, enzyme-linked immunosorbent assay (ELISA)-based and real-time polymerase chain reaction (PCR) tests, using amplification of the same segment of the CMV genome. The aim of this study was to compare sensitivity, specificity, and laboratory performance characteristics of these assays. In both assays, a 159-bp segment of UL83 gene was amplified. External and internal controls were constructed by cloning the amplification product and heterogenous DNA segment flanked by target sequences for CMV-derived primers into bacterial plasmids, respectively. Real-time PCR was performed on LightCycler (Roche Diagnostics, Mannheim, Germany), and amplicons were detected using fluorescence resonance energy transfer probes. Alternatively, PCR products were labeled by digoxigenin, hybridized to immobilized probes, and detected by ELISA. The assays were tested on genomic DNA isolated from laboratory strains of CMV, QCMD control panel, and CMV DNA-positive peripheral blood DNA samples from hematopoietic stem cell transplant recipients, previously characterized by pp65 antigenemia and qualitative nested PCR. Real-time and ELISA-based PCR assays showed a linear course of 1-10(8) and 10-10(5) copies of CMV DNA per reaction, respectively. When compared with ELISA-based PCR, real-time PCR showed superiority in inter- and intra-assay reproducibility. Both assays were highly specific in detecting CMV DNA. No difference in amplification efficiency of internal or external standards and wild-type CMV DNA was found. The assays exhibited 83% concordance in CMV DNA detection from clinical samples, all discrepant samples having low CMV DNA copy numbers. There was a good correlation between viral DNA loads measured by the 2 assays. Statistically significant correlation was observed between the numbers of CMV DNA copies and pp65-positive leukocytes in the samples tested. Both variants of competitive PCR are adequately sensitive to be used for CMV DNA quantitation in clinical samples. LightCycler PCR, having superior performance characteristics and being less time-consuming, seems to be more suitable for routine diagnosis.  相似文献   

10.
Human cytomegalovirus (CMV) is a well-known cause of morbidity and mortality in transplantation patients. Monitoring of CMV reactivation from latency is critical for these patients. The key to efficient and effective management of CMV infection is a test capable of rapidly monitoring and quantifying the presence of CMV in the blood. This is essential for the identification of subjects at high risk of developing CMV disease, for example, patients receiving steroid or immunosuppressive compounds for accelerated graft-versus-host disease, transplant rejection and also for the application and monitoring of pre-emptive antiviral therapeutic strategies. The assays presently available and frequently used in this setting include conventional and shell vial culture, the CMV antigenemia assay, PCR for CMV DNA, hybrid capture assay for CMV DNA and detection of CMV RNA by nucleic acid sequence-based amplification. The low sensitivity and low reproducibility of conventional cell culture and shell vial assays limit their role in the management of CMV infection to one of disease diagnosis. Diagnostic assays, such as the pp65 antigenemia and other molecular assays, have improved the ability to diagnose CMV disease quickly and accurately. These methods fulfill the requirements for a good diagnostic assay: they have high sensitivity, most can quantify viral load and they are rapid and reproducible. Their characteristics allow these assays to be used to predict the development of CMV disease and monitor response to therapy.  相似文献   

11.
背景:异基因造血干细胞移植后EB病毒再激活可导致致死性的移植后淋巴细胞增殖性疾病及相关疾病,目前国内尚未建立完整的EB病毒再激活及其相关性疾病的诊疗体系。目的:前瞻性研究异基因造血干细胞移植后患者EB病毒再激活的发生和相关危险因素。方法:纳入129例接受异基因造血干细胞移植的患者,采用实时定量PCR方法定期测定外周血中EB病毒载量,Kaplan-Meier模型分析其再激活的发生率,Logistic回归分析模型分析其再激活的相关危险因素。结果与结论:异基因造血干细胞移植后EB病毒再激活及其相关疾病发生率高,EB病毒再激活发生的危险因素有HLA配型不合、应用抗胸腺细胞球蛋白、Ⅲ~Ⅳ度急性移植物抗宿主病及年龄小于20岁。  相似文献   

12.
异基因造血干细胞移植后发生巨细胞病毒感染的临床分析   总被引:1,自引:0,他引:1  
本研究总结巨细胞病毒(CMV)感染的发生及治疗。对北京军区总医院血液科2010年1月至2012年1月140例异基因造血干细胞移植(allo-HSCT)患者的临床资料进行了回顾性分析。结果表明,140例allo-HSCT中48例患者发生移植后CMV感染,发生率为34.3%,首次检出CMV-DNA阳性中位时间为移植后45(33-68)d,CMV定量范围为1.25×103-5.5×106,其中2例为CMV相关性间质性肺炎,5例为CMV相关性出血性膀胱炎。发生移植物抗宿主病(GVHD)患者共65例,其中合并CMV感染32例,占49.2%。应用更昔洛韦、膦甲酸钠抗CMV治疗45(33-68)d后CMV-DNA转阴,有效率为100%。共有12例患者治疗过程中出现一过性的白细胞和血小板减少。结论:allo-HSCT后较易发生CMV感染,发生GVHD的患者CMV感染发生率也较高,更昔洛韦、膦甲酸钠对allo-HSCT后CMV感染的治疗效果可靠,且不良反应少。  相似文献   

13.
目的:分析异基因造血干细胞移植(allo-HSCT)后并发出血性膀胱炎(HC)的危险因素。方法:回顾性分析2010年1月-2018年12月于西安交通大学第一附属医院行allo-HSCT的153例患者的临床资料。观察HC的发生率、发生中位时间及治疗转归。利用多因素分析观察患者性别、年龄、诊断、移植前疾病状态、移植类型、预处理方案中是否含有ATG、预处理方案中是否含有CTX、干细胞来源、中性粒细胞植入时间、血小板植入时间、CMV感染、EBV感染、BKV感染、急性移植物抗宿主病(a GVHD)是否为发生HC的高危因素。结果:153例allo-HSCT患者中,25例发生HC,发生率为16.34%,发生的中位时间为31 d,治疗后全部好转,无1例遗留膀胱刺激症状及膀胱挛缩。经单因素和多因素Logistic回归分析,结果显示,移植类型、预处理中含ATG、CMV血症以及a GVHD 4项因素(R值分别为1.036、3.234、3.298、2.817)是HC的独立危险因素。结论:HC患者尿BKV检测均为阳性,主要发生在移植后13-56 d。HLA配型半相合、含ATG的预处理方案、CMV血症及a GVHD是allo-HSCT后发生HC的独立危险因素。  相似文献   

14.
目的 比较血pp6 5抗原和IgM抗体检测对移植患者巨细胞病毒 (CMV)活动性感染的诊断价值。方法 收集 5 8位移植患者的血标本 (共 2 0 8份 ) ,分离血浆和多形核白细胞 ,血浆用于CMVIgM抗体检测 (ELISA) ,白细胞用于pp6 5抗原血症检测(荧光免疫组化 )。同时随机取部分血浆标本进行荧光定量PCR分析 ,检测CMVDNA。结果 pp6 5抗原阳性细胞数与CMVDNA拷贝数呈正相关 (r=0 .87) ,也与病人的临床症状密切相关 ,可用于监测移植患者CMV的活动性感染。IgM抗体与pp6 5抗原阳性的一致率为 2 1.2 % ,对CMV感染的敏感性为 2 2 .3% ,特异性为 95 .6 %。结论 pp6 5抗原阳性细胞数能反映CMV的数量 ,可监测CMV活动性感染 ,检测方法特异、灵敏、操作简单 ,适宜临床实验室应用。血清IgM抗体检测因为敏感性较低 ,不适用于移植患者CMV活动性感染的监测。  相似文献   

15.
为了研究ABO血型不合异基因造血干细胞移植(allo—HSCT)后并发纯红细胞再生障碍(pure red cell aplasia,PRCA)的发病情况及危险因素,对本医院以往血型不合异基因造血干细胞移植进行回顾性分析。探讨移植后患者PRCA的发病危险因素。研究结果表明,72例ABO血型不合allo—HSCT患者中,4例发生PRCA,其中A供O3例,A供B1例。PRCA的发生不影响急性移植物抗宿主病(GVHD)或巨细胞病毒(CMV)感染的发生。PRCA患者红系恢复的时间显著长于未PRCA发生患者。结论:PRCA是ABO血型不合移植的主要并发症。A供O可能是ABO血型不合allo—HSCT后并发PRCA的危险因素。  相似文献   

16.
We studied the recovery of CMV-specific CD4+ and CD8+ T-cell immunity in 52 recipients of allogeneic stem cell transplantation (SCT). The proportions of IFN-gamma-producing CD4+ and CD8+ T cells upon in vitro activation using peptide pools representing the CMV pp65 and IE-1 proteins were assessed at multiple time points post SCT, and correlated with the occurrence of CMV reactivation. In a retrospective analysis, recurrent CMV reactivations occurred in 9 patients and were associated with low pp65-specific CD4+ T-cell and low IE-1-specific CD8(+) T-cell reactivities, whereas patients without detectable CMV reactivation (n = 30) or a single reactivation (n = 13) showed a better recovery of these immune responses. CD4+ T-cell responses to IE-1 were infrequent in most patients, whereas CD8+ T-cell responses to pp65 occurred frequently, but did not correlate with protection against (recurrent) reactivation. Prospectively, CMV-specific T-cell responses could be studied prior to 14 reactivation episodes in 8 patients. CD4+ T-cell responses to IE-1 and pp65 were positive in only 1 and 2 episodes, respectively. CD8+ T-cell responses against IE-1 were positive in 4, but against pp65 in 12 episodes, again showing that CD8+ T-cell reactivity against pp65 did not prevent CMV reactivation. Thus, monitoring of particular CMV-specific CD4+ and CD8+ T-cell responses after allogeneic SCT may identify patients at risk for recurrent CMV reactivations.  相似文献   

17.
目的用实时荧光定量聚合酶链反应(FQ-PCR)和pp65抗原检测儿童人巨细胞病毒(HCMV)感染,并对其诊断HCMV活动性感染进行比较评估。方法分析924例HCMV血清学检测阳性的儿童的全血HCMV-DNA荧光定量PCR及HCMV-pp65抗原检测结果,与临床诊断的符合程度进行比较。结果全血HCMV-DNA荧光定量PCR和pp65抗原检测的一致率为81.17%,其标准误为0.39,Kappa值为0.60;PCR检测HCMV感染的灵敏度为98.4%,特异度为97.3%,Youden指数为0.957,PVP和PVN分别为0.961和0.989;pp65抗原检测用于检测HC-MV感染的灵敏度为59.2%,特异度为100%,Youden指数为0.592,PVP和PVN分别为1.00和0.782。结论全血HCMV-DNA荧光定量PCR和pp65抗原检测的结果有较好的一致性,诊断HCMV感染PCR检测的灵敏度较pp65抗原检测的灵敏度要高,而后者的特异度较前者高。二种方法都可用于CMV活动性感染的诊断。  相似文献   

18.
BACKGROUND: Cytomegalovirus (CMV) disease constitutes a serious complication after allogeneic stem cell transplantation. For the clearance of CMV, CD8+ T cells are pivotal. STUDY DESIGN AND METHODS: Here, the novel streptamer technology was used at good manufacturing practice (GMP) level for adoptive transfer of CMV‐specific T cells into acute leukemia patients with recurrent high CMV antigenemia after allogeneic stem cell transplantation. RESULTS: After a single transfusion, the frequency of CMV‐specific CD8+CD45RA+CCR7– effector T cells increased dramatically from 0.0% to a maximum of 27.1% of all T cells. These T cells were clearly donor derived and did not stem from intrinsic reconstitution, as demonstrated by analysis of 1) donor chimerism through single‐tandem repeats, 2) T‐cell receptor excision circles, and 3) Vβ‐chain typing by polymerase chain reaction. Clinically, the specific T‐cell transfer resulted in a persistent clearance of the CMV antigenemia, which allowed the patients to discontinue toxic antiviral drug therapy without further high‐level reactivation of CMV, demonstrating the power of the streptamer technology. CONCLUSION: Taken together, the streptamer technology offers the advantage of selecting virus‐specific CD8+ T cells at GMP level for adoptive T‐cell transfer, thus inducing long‐lasting specific CD8+ T‐cell responses without increasing the risk for graft‐versus‐host disease.  相似文献   

19.
目的建立实时荧光PCR方法以检测广谱人乳头瘤病毒。方法该实时荧光PCR方法利用了degenerate PCR引物与非特异性的SYBR Green染料替代通常使用的PCR特异性引物与探针,并与市售高危13型、低危6型试剂盒以及degener-ate PCR方法进行比较。结果对于用高危13型试剂盒及低危6型试剂盒检测的100例黏膜型人乳头瘤病毒阳性标本,用SYBR Green实时荧光PCR方法检出的阳性数为69例,敏感性为69%,相反,用SYBR Green实时荧光PCR方法检测出的40例皮肤型人乳头瘤病毒阳性标本,用高危13型试剂盒以及低危6型试剂盒检测均为阴性;对于用SYBR Green实时荧光PCR方法及degenerate PCR方法检测的90例黏膜型、60例皮肤型标本,其检测结果显示两种方法有高度一致性(Kappa=0.907)。结论与市售试剂盒相比,SYBR Green实时荧光PCR方法可以覆盖广谱的HPV型别(皮肤型与黏膜型),并且比degenerate PCR方法高效、快速和简便,不失为一个可用于临床与科研的良好检测手段。  相似文献   

20.
异基因造血干细胞移植(allo—HSCT)是治疗骨髓增生异常综合征(MDS)的有效手段,MDS发病率随年龄增加而增加,尽管近年来传统清髓性移植治疗MDS取得可喜的成绩,但对于年龄大于60岁的MDS患者,allo—HSCT预后差,低强度预处理异基因造血干细胞移植(RIC allo—HSCT)使老年MDS患者进行allo—HSCT成为可能,利用供者淋巴细胞的移植物抗肿瘤作用显著降低移植相关器官毒性及非复发死亡率.本文就RIC—HSCT治疗MDS的问题诸如RIC allo—HSCT治疗MDS可行性,MDS病例选择,RIC allo—HSCT的时机干细胞来源,RIC预处理方案.疗效及预后评价.GVHD与移植物抗MDS效应等的研究进展做一综述及展望。  相似文献   

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