器官移植术后人类巨细胞病毒感染的诊断

外周血白细胞 (ＷＢＣ)中人类巨细胞病毒 (ＨＣＭＶ)抗原的出现称之为抗原血症 ,我们采用ＥｎｖｉｓｉｏｎＴＭ 免疫组化法检测外周血白细胞中ＨＣＭＶ抗原从而诊断ＨＣＭＶ感染 ,达到了早期、快速的目的 ,并指导临床合理治疗。研究对象为广州地区 12 2例器官移植术后受者 ,其中肾移植 86例 ,骨髓移植 2 0例 ,肝移植 12例 ,脐血造血干细胞移植 4例。移植术后 2周起 ,每周采血 1次 ,2ｍｌ 次。共采集抗凝外周全血 32 8份。实验用试剂ＨＣＭＶ即刻早期、早期单克隆抗体 (ＤＤＧ9+ＣＣＨ2株 ) ,免疫组化ＥｎｖｉｓｉｏｎＴＭ 试剂盒均购自丹麦…     

肝移植术后早期巨细胞病毒感染的先驱性治疗

目的 通过临床病例对照研究,寻求肝移植术后防治巨细胞病毒(CMV)感染的较好方法。方法 将63例原位肝移植患者分为预防性治疗组与先驱性治疗组,术后3个月内定期进行CMV-PP65定性和CMV-DNA定量检测,预防性治疗组均在术后2周时给于静脉更昔洛韦治疗,先驱性治疗组仅在检测阳性时给予更昔洛韦治疗。结果 预防性治疗组中17％(5／35)出现了CMV感染;先驱性治疗组中36％(10／28)出现了CMV感染。两组中全部病例均未发生CMV病。结论 肝移植术后早期采用先驱性治疗不增加巨细胞病毒病的发生率。     

HBV感染对肝移植术后巨细胞病毒感染的影响

目的 探讨肝移植患者移植术前后HBV感染对移植术后巨细胞病毒(CMV)感染及其肝功能的影响.方法 回顾性分析我院近两年来113例肝移植患者临床资料,排除术后1月内死亡、血型不符、失访和并发其他噬肝病毒感染患者,共有102例患者纳入.根据肝移植术前是否感染HBV将患者分为HBV感染组和对照组(非HBV感染);对于感染组患者,又根据移植术后HBV是否复发分为术后复发组和非复发组,采用免疫组化方法 检测CMV被膜抗原pp65水平,时间分辨法检测乙肝病毒抗原和抗体水平,常规方法 检测肝转氨酶水平.结果 HBV感染组(n=83)和对照组(n=19)中,术后pp65阳性率分别为84.3%(70/83)和57.9%(11/19)(P=0.024);HBV感染组中,HBV复发组和非复发组中pp65阳性率分别为90.9%(10/11)和83.3%(60/72)(P=0.843),各组间肝转氨酶水平差异无统计学意义(P0.05).结论 肝移植术前HBV感染可以增加术后CMV病毒血症风险,而肝移植后HBV复发并不增加CMV病毒血症风险;同时,与单一HBV或CMV感染而言,两种病毒联合感染并不明显引起肝转氨酶水平的升高.     

孕妇巨细胞病毒感染后的治疗分析

人巨细胞病毒 (ＨＣＭＶ)在人群中感染广泛 ,对人类的危害 ,尤其对胎儿的危害严重 ,孕期发生ＨＣＭＶ原发感染或复发感染均可经胎盘或产道感染胎儿 ,可导致早期流产、死胎、新生儿死亡及活产婴儿先天性畸形。部分病例出生时可无异常 ,但随着年龄的增长 ,逐渐表现出语言障碍、智     

流式细胞术在巨细胞病毒感染诊断中的应用

巨细胞病毒(cytomegalovirus, CMV)感染是器官移植受者、AIDS患者和其它严重免疫抑制患者的常见并发症之一，也是优生优育监测的重要指标〔1〕。CMV早期即刻抗原(immediate early antigen IEA)是GMV感染的最客观依据之一，其检测方法主要有离心培养法(centrifuged culture)〔2〕和免疫组织化学技术等。     

巨细胞病毒感染在抽动障碍中的临床意义初探

目的：探讨人巨细胞病毒（human cytomegalovirus,HCMV)感染在抽动障碍中的临床意义。方法：应用PCR基因扩增技术对66例抽动障碍患儿进行血液HCMV检测,并测定74例正常儿童作为对照,结果：抽动障碍患儿HCMV检出阳性率（26％）明显高于对照组（3％）,差异有显著性（P＜0．01）,抽动障碍三种类型间HCMV感染阳性率无显著性差异（P＞0．05）。结论：HCMV感染与抽动障碍发病有关。     

肝移植术后受者巨细胞病毒感染PBMC中干扰素γ和IL-4阳性细胞的变化

巨细胞病毒（CMV）感染是器官移植术后的一种常见并发症，CMV感染率在所有实质性器官移植病人中约为20％～60％，CMV发病的危险性在移植术后第2～3个月最高。TH1和TH2细胞因子平衡的改变与许多免疫相关疾病的转归和预后相关，也与CMV感染的急性反应、播散、活化及逃避宿主的免疫清除等致病性密切相关，因此本文通过检测干扰素γ（IFN-γ）和白细胞介素4（IL-4）水平，探讨其与CMV感染发病的关系。     

妊娠期巨细胞病毒感染

巨细胞病毒是引起新生儿先天性感染最常见的病原微生物之一,本文介绍了孕妇巨细胞病毒感染情况及其感染对妊娠结局、胎儿和婴儿的影响,并对妊娠期感染巨细胞病毒防治作了简要介绍。     

流式细胞仪检测巨细胞病毒感染

巨细胞病毒(CMV)是巨细胞包涵体病的病原体, 是疱疹病毒家族的DNA病毒, 具有典型的疱疹病毒结构.人群中CMV感染非常广泛, 60%-90%的成人已有CMV抗体.     

聚合酶链反应技术在诊断孕妇人巨细胞病毒感染中的应用

人巨细胞病毒 (humancytomegalovirus,HCMV)是引起宫内感染的常见病原体之一。孕妇感染HCMV后 ,可通过胎盘垂直传播 ,引起流产、早产、死胎、胎儿畸形或新生儿先天性感染。为了早期、快速地诊断HCMV感染 ,为临床治疗或终止妊娠提供依据 ,降低畸胎或缺陷患儿的出生 ,达到优生优育的目的 ,已经建立了许多用于诊断HCMV感染的技术 ,如血清学法、病毒培养法、病毒血症检测法、抗原血症检测法及聚合酶链反应法 (polymerasechainreaction ,PCR)等。其中 ,PCR是一种DNA体外引物定向酶促扩增技术 ,具有简便、敏感和特异的优点。目前 ,采用…     

494例次移植肝穿刺活检病理组织学分析

目的 通过对354例(494例次)移植肝穿刺活检组织进行病理分析，观察移植肝的组织学变化，探讨其出现肝功能不全的原因。方法 移植肝穿刺活检组织经10％中性福尔马林固定，快速石蜡连续切片，常规HE染色。部分病例做VG、Masson、PAS、网状纤维组织化学和免疫组织化学染色，抗体为HBsAg、HBcAg、HcVAg、CMV、CD8、CD4、CK19。对排斥反应病例，依照国际统一的BANFF标准进行急性排斥反应分级，应用排斥活动指数(RAI)进行排斥反应程度评分。结果 急性细胞性排斥反应最常见，180例(50．85％)，慢性排斥反应11例(3．11％)，再灌注缺血损伤20例(5．65％)，胆汁淤滞及急慢性小胆管炎64例(18．08％)，药物性肝损害18例(5．08％)，移植肝无功1例(0．28％)，CMV感染24例(6．78％)，乙肝病毒再感染及乙肝复发27例(7．63％)，丙型肝炎复发2例(0．56％)，原发性硬化性胆管炎复发1例(0．28％)，难以诊断6例(1．69％)。结论 移植肝穿刺活检对移植术后并发症的诊断及选择治疗方案具有重要价值。     

Virological and serological diagnosis of cytomegalovirus infection in bone marrow allograft recipients

To detect cytomegalovirus (CMV) infections, a total of 1,074 cultures of urine, saliva, or blood were collected weekly from 43 consecutive patients undergoing allogeneic bone marrow transplantation. Twenty-three patients were seronegative before transplant and primary infection occurred in 2 (9%). Twenty patients were initially seropositive and recurrent infections occurred in 5 (25%). Three patients in the recurrent group had proven CMV pneumonitis; viraemia was detected in two recipients, while the third had CMV isolated only from bronchial lavage fluid. The serological response of the 43 patients was defined by testing 559 serial sera for specific IgG and IgM antibodies by radioimmunoassay. Passive acquisition of IgG antibodies from blood products was found in 78% of initially seronegative recipients. One patient with primary infection responded in a pattern typical of immunocompetent individuals with long-term production of specific IgG and transient production of specific IgM antibodies. The second patient also had a typical response, but this was delayed until several weeks after the start of virus excretion. In patients with recurrent infections, specific IgM production did not correlate with episodes of virus excretion. Three of five such patients failed to mount a specific IgM response, and these were the only patients in the study to develop CMV pneumonitis. We conclude that CMV infection in bone marrow recipients can only be diagnosed by detection of virus; therefore, the ability of these patients to mount humoral immune responses should not be relied upon for diagnostic purposes.     

High frequency of gastroduodenal cytomegalovirus infection in liver transplant patients

The prevalence and significance of cytomegalovirus (CMV) detected in biopsy specimens from the gastroduodenal mucosa of liver transplant patients, patients with chronic or acute liver failure and immunocompetent patients with dyspeptic symptoms were evaluated. 80 liver transplant patients with upper gastrointestinal symptoms, 132 patients with chronic and 25 with acute liver failure, and 33 immunocompetent, dyspeptic patients underwent oesophagogastroduodenoscopies, with biopsies from the duodenum and stomach. CMV was demonstrated by immunohistochemistry in frozen sections, using a monoclonal antibody against CMV-specific antigens (pp65 matrix protein), and in paraffin sections by a monoclonal antibody against delayed early protein (p52). 71% of the liver transplant patients, 45% of the patients with chronic liver disease, 20% with acute liver failure, and 45% of the immunocompetent, dyspeptic patients had CMV-positive findings in the gastroduodenal mucosa (liver transplant patients vs other groups, p<0.01). Histopathological findings in CMV-positive samples were focal inflammation, including increased inflammation of the lamina propria, infiltrating leukocytes intra-epithelially, regenerative changes in the epithelial cells and inclusion bodies. In conclusion, CMV-positive cells and inclusions are often found in the gastroduodenal mucosa of liver transplant patients, as well as in patients suffering from chronic liver disease or even in otherwise healthy patients with dyspeptic symptoms.     

Presence of occult cytomegalovirus infection in the brain after orthotopic liver transplantation

Cytomegalovirus (CMV) infection remains a highly prevalent systemic complication following orthotopic liver transplantation (LT), accounting for a significant increase in morbidity and affiliated costs. However, unlike other immunosuppressed groups of population, CMV infection of the central nervous system in LT is rarely diagnosed, either clinically or postmortem. Furthermore, in 20% of the LT patients who develop preterminal neurological complications, the etiology remains undetermined. With the hypothesis that at least some of these cases could be related to an occult CMV infection, we examined brain tissue from 83 unselected autopsies of LT patients by morphological, immunohistochemical (IHC), in situ hybridization (ISH), and nested polymerase chain reaction (nested PCR) techniques. Microglial nodules were observed in 17 brains of the LT group (20.4%) but in none of the 36 controls. Isolated positive cells by either IHC, ISH, or both techniques, were identified in 11 LT patients (13.2%) and in 2 controls (5.5%). CMV DNA amplification was obtained from paraffin-embedded tissues in 41 of 81 LT cases (50.6%), and in 5 controls (13.8%) (P=0.00017). Viral inclusion bodies, inflammatory infiltrates, or necrotizing changes were not identified in any case. Our findings indicate an increased susceptibility of the brain of LT patients to occult infection by CMV and suggest that a latent or low-grade infection of the central nervous system could operate as a reservoir of the CMV and play a role in some of the unexplained neurological symptoms that appear in the postoperative period.     

Transfusion practice in orthotopic liver transplantation

Liver transplant procedures require the most blood components, despite the fact that blood use in liver transplantation has declined dramatically over the last decade. Liver transplant recipients present unique challenges, not only in terms of blood supply, but also requirements for specialized blood components, serologic problems, and immunologic effects of transfusion on both the allograft and the recipient. The cause of intraoperative blood loss in liver transplantation is multifactorial, due to both technical factors and poor coagulation control. This procedure carries the risk of massive blood loss, which requires massive transfusions and is associated with postoperative infections, reduced graft survival, multi-organ dysfunction, and higher risk of mortality. Efforts to reduce intraoperative bleeding leading to limitation of blood transfusions are desirable to improve results and also to control costs.Method of literature search:

1. The name of topic is typed and searched in Google search.
2. The name of topic is typed and searched in PubMed search. Related articles were also searched.
3. Some standard books in Transfusion Medicine were also referred.

     

A serological investigation of human herpesvirus 6 infections in liver transplant recipients and the detection of cross-reacting antibodies to cytomegalovirus.

Sera from 50 orthotopic liver transplant recipients were examined for antibodies to human herpesvirus 6 (HHV-6) and cytomegalovirus (CMV), and the findings correlated with the clinical condition of the patients. Both primary and secondary HHV-6 infections were detected serologically following liver transplantation. Interpretation of serological assays is complicated by CMV and HHV-6 antibody cross reactions which were common. Sera from 5 patients became HHV-6 antibody negative following absorption with CMV infected cells. Thirty patients were initially seronegative for HHV-6 antibodies, 12 remained so following transplantation, 5 developed cross reacting antibodies, and 13 seroconverted. The seroconversions occurred at 4 to 8 weeks post-transplant in the same time period as CMV antibody rises. HHV-6 IgM was detected in only 4 of the 13. Of the 7 patients who had serological evidence of active HHV-6 infections but no evidence of CMV infection, 4 (56%) had fever, 1 (14%) hepatitis, 1 (14%) lung dysfunction, and 3 (42%) neurological disorders. In the 12 patients who remained HHV-6 antibody negative, there were fewer fevers and neurological disorders.     

Comparison of two methods used for monitoring low-copy cytomegalovirus infection in a patient with chronic myeloid leukemia after unrelated umbilical cord blood transplantation

Introduction:

Detection of human cytomegalovirus (CMV, HHV-5) DNA in clinical specimens is considered a cornerstone in the diagnosis of HHV-5 disease. The present study compared two quantitative methods used for diagnosing cytomegalovirus infection in a 21-year-old woman with chronic myeloid leukemia after an unrelated umbilical cord blood transplantation.

Materials and Methods:

Blood samples were tested for the presence of HHV-5 DNA using the LightCycler PCR, the quantitative Eclipse^® CMV DNA Detection Kit, and a qualitative in-house PCR assay using primers that amplify part of the HHV-5 MIE gene.

Results:

Results from samples containing a low cytomegalovirus load were more accurate with the LightCycler test than those obtained with the Eclipse^® test, which underestimated the viral load of samples containing low DNA copy numbers.

Conclusions:

These findings underline the value of novel PCR methods used in current therapeutic procedures and in monitoring antiviral therapy with nucleoside analogs. The high level of sensitivity, specificity, accuracy, and rapidity provided by the LightCycler instrument are favorable for the use of this system in the detection of HHV-5 DNA in clinical specimens.

     

Hepatic reinnervation following orthotopic liver transplantation in man.

We have studied changes in the pattern of intrinsic hepatic innervation in sequential liver biopsies from 16 patients who underwent orthotopic liver transplantation. Seventy-one needle biopsies were used, including specimens obtained at the time of transplantation (time zero) and up to 4 years post-transplantation; five transplant hepatectomy tissue blocks removed 3-32 months after transplantation were also assessed. Paraffin sections were immunostained with anti-PGP 9.5 and anti-S-100 to identify nerve fibres. All 'time zero' biopsies contained portal nerves and all but two showed staining of parenchymal fibres. After 1 week, no subsequent biopsies contained parenchymal fibres. The disappearance of portal fibres was less rapid and showed greater variability between patients, but they had all disappeared by 6 weeks and there was no positive staining between 6 and 60 weeks. Thereafter, a minority of biopsies showed innervation of a few small portal tracts. Samples from the porta hepatis, hepatectomy specimens, and needle biopsies containing large tracts showed persistence of major nerve trunks at all stages. Abnormally large nerve bundles were seen in some of these areas. The pattern of nerve staining showed no obvious relationship to the intensity of rejection changes. Our results suggest that there is a limited, delayed capacity for regeneration of portal, but not parenchymal, fibres in the transplanted human liver. The physiological significance of this long-term parenchymal denervation in transplanted livers remains to be determined.