肾移植患者微小病毒B19感染的实验研究

目的：研究肾移植术后微小病毒B19感染的情况。方法：采用间接ELISA的方法检测肾移植术后患者血清标本中抗微小病毒－B19（HPV－B19）IgM和IgG抗体的浓度,并与肾移植前及健康献血员进行比较。结果：肾移植后患者血清标本中HPV－B19IgM抗体的浓度和阳性率明显高于肾移植前和健康对照,而HPV－B19IgG抗体无明显改变。结论：肾移植术后,机体免疫力下降,HPV－B19感染的机率上升,可能引起以贫血为主的多系统多器官的功能障碍,影响肾移植的效果。     

某部急性上呼吸道感染暴发疫情的病原学检测

目的 通过实验室病原学筛查,结合流行病学史及临床表现,初步筛查某部急性上呼吸道感染暴发疫情的致病病原体.方法 采集51例患者血液和咽拭子标本,分别采用免疫荧光、酶联免疫吸附试验(ELISA)、聚合酶链反应(PCR)、病毒分离及微生物学细菌培养等实验室诊断方法筛查、确定致病病原体,并对15例恢复期患者(发病后15～28d)血清进行IgG滴度检测,比较急性期与恢复期抗体滴度的变化.结果 所有咽拭子的细菌培养及7例咽拭子的病毒分离试验结果均为阴性.对24例患者血清进行的免疫荧光法呼吸道常见20种病原体IgM抗体筛查结果显示:流感A型阳性率为20.8%(5/24),流感B型阳性率为12.5%(3/24),肺炎衣原体阳性率为20.8%(5/24),肺炎支原体阳件率为50.0%(12/24),其余病原体检测均为阴性.ELISA检测IgM抗体结果显示:总流感病毒IgM抗体阳性2例(3.92%),肺炎衣原体抗体阳性7例(13.7%),肺炎支原体抗体阳性20例(39.2%),呼吸道合胞病毒抗体阳性2例(3.92%),EB病毒抗体阳性1例(1.96%).免疫荧光法肺炎支原体IgG抗体滴度检测结果显示1:100稀释后阳性反应37例(72.5%),其中的31例(60.8%)在1:1000稀释后仍呈阳性反应,对照组仪2例(6.45%)IgG抗体1:100时呈阳性,在1:1000稀释后均为阴性.PCR实验未见特异扩增反应.恢复期与急性期血清抗体效价检测发现15例中有7例肺炎支原体抗体效价有4倍以上变化.结论 初步推断此次某部群体急性上呼吸道感染事件系肺炎支原体感染引起;该疫情的实验室快速诊断为今后疫情的早期诊断提供了思路.     

新兵人群庚型肝炎病毒感染流行病学调查

目的：了解新兵人群庚型肝炎病毒感染情况。方法：采用血清流行病学调查方法,利用酶联免疫吸附试验（ELISA）技术,随机抽取近5年入伍新兵444例血清进行庚肝病毒IgM和IgG抗体检测,统计分析其阳性率。结果：各年度新兵血清庚肝病毒IgM、IgG均未检出;对2006、2007年入伍新兵9169例筛查出的HBsAg阳性20例标本,进一步行庚肝病毒IgM、IgG检测均为阴性。结论：新兵人群中庚型肝炎病毒既往感染率较低。     

肺泡灌洗液联合肺组织PCR方法检测结核杆菌DNA对肺结核早期诊断的意义

目的探讨肺泡灌洗液(BALF)联合肺组织聚合酶链反应(PCR)方法检测结核杆菌DNA对肺结核早期诊断的意义。方法对104例结核杆菌痰阳性和确诊肺结核的痰菌阴性患者经纤维支气管镜采集BALF及活检肺组织进行石蜡包埋,对标本采用PCR方法检测结核杆菌DNA。结果 104例患者BALF中PCR检测结核杆菌DNA阳性92例,阳性率为88.46%;肺组织石蜡包埋检测结核杆菌DNA阳性98例,阳性率为94.23%。两组阳性率比较,差异无统计学意义(P>0.05)。结论 BALF联合肺组织PCR方法检测结核杆菌DNA有较高的敏感性及特异性,对肺结核病的早期诊断有重要临床意义。     

HLA-G5在健康人和肝肾移植受者体内的表达分析

目的了解国内健康人和器官移植受者体内人类白细胞抗原-G5(HLA-G5)的表达情况,并探索其表达的时间规律。方法分别采集30例健康人(健康查体正常)、50例肝移植受者(肝移植术后3个月且肝功能稳定)、50例肾移植受者(肾移植术后3个月且肾功能稳定)的外周血各3ml,并采集33例肝肾移植受者术前及术后1周、4周、12周、1年的外周血各3ml。提取血清,采用酶联免疫法(ELISA)检测HLA-G5水平。结果30例健康人中,28例HLA-G5的A值低于0.5,根据标准曲线含量为0.0ng/ml,2例分别为8、9ng/ml。50例肝移植受者HLA-G5表达阳性者16例,阳性率为32%,其中4例含量高于30ng/ml。50例肾移植受者HLA-G5表达阳性者10例,阳性率为20%,有1例含量为25ng/ml。健康人、肝移植、肾移植3组血清中HLA-G5平均含量分别为0.56±0.20、8.34±1.50和3.26±0.25ng/ml。对33例肝肾移植受者进行动态检测显示,有1例患者术前和术后1周HLA-G5呈阳性,4例术后4周阳性,12例术后12周阳性,追踪至术后1年有11例阳性。结论HLA-G5在健康人体内低表达,与器官移植免疫耐受有一定相关性。在移植后移植物接受良好的情况下,HLA-G5有不同水平的表达,肝移植受者的HLA-G5表达量高于肾移植受者。33例肝肾移植受者的动态结果证实了HLA-G5的表达从mRNA转录到蛋白表达的时间为15~60d的观点,HLA-G5的表达在第60天到达高峰,并具有稳定性。     

聚合酶链反应检测人巨细胞病毒的方法及临床应用

在人巨细胞病毒（HCMV）高度保守区IEA基因内设计一对引物，建立了聚合酶链反应（PCR）检测HCMVDNA的方法，经实验证明有高度的特异性和敏感性，对其它疱疹病毒和正常人DNA无交叉反应，HCMVAD169株DNAEcoRⅠ酶切J片段的最小检出量为0.1fg,相当于6个基因拷贝，对PCR反应体系中的有关实验条件如Mg^2 浓度，引物浓度，TaqDNA多聚酶的浓度，循环程序等及尿标本的处理方法进行优化选择，用PCR检测20例肾移植受尿标本中HCMVDNA，结果11例（55%）阳性，表明PCR是一种快速，特异，敏感的检测方法，适用于肾移植等病人的HCMV感染监测。     

RF对ELISA法检测病毒性肝炎IgM类抗体的影响

目的:研究RF对ELISA法检测病毒性肝炎IgM类抗体的影响,为临床检测中出现假阳性结果的分析提供依据。方法:选择用EHSA法测得的病毒性肝炎IgM类抗体阳性标本,用速率散射法测其RF,中和RF处理后重测IgM抗体。结果:5组标本(抗HAV-IgM阳性组、抗HDV-IgM阳性组、抗HBc-IgM阳性组、抗HAV-IgM与抗HDV-IgM均阳性组、抗HBc-IgM与抗HDV-IgM均阳性组)由RF引起的假阳率分别为15.4％、20.0％、15.0％、25.0％、43.8％。高RF浓度组引起的假阳性率为78.8％,低RF浓度组引起的假阳性率为15.4％。结论:RF对5组标本的检测均有干扰作用,其干扰率无显著性差异。高浓度RF组干扰率比低浓度RF组大。     

不明原因呼吸道感染疫情病原学分析

目的 对某部营区内的群发上呼吸道感染进行病因学研究,以明确诊断,控制并预防疫情扩散.方法 采集32例住院患者的血清和咽拭子标本,其中4例有急件期及恢复期双份血清.血清标本采用欧蒙间接免疫荧光法(IFA)和ELISA法进行20种呼吸道相关病原IgM抗体筛查,双份血清进行滴度比较;咽拭子标本进行病毒分离培养和细菌培养.依据血清学结果对咽拭子进行相关病原DNA的提取、扩增及测序分析.结果 血清ELISA检测结果显示32例患者中腺病毒IgM阳性12例,肺炎衣原体IgM阳性15例.TFA检测结果显示肺炎衣原体kM阳性17例,肺炎支原体、流感A病毒、副流感病毒IgM阳性各1例.4例双份血清中肺炎衣原体和腺病毒IgM各有2例滴度呈4倍变化.细菌培养及病毒分离培养均阴性.1例高热患者咽拭子标本3对腺病毒相关引物PCR呈阳性,测序显示为腺病毒11型.采用大环内酯类抗生素进行治疗,大部分病例有效.结论 结合临床特征及病原学分析结果,判断此次疫情的原因主要为腺病毒及肺炎衣原体的合并或交替感染.     

乙型肝炎病毒血清标志物与PreS1抗原的关系

目的：了解PreS1与HBVM和HBV DNA的关系及其临床应用价值。方法：对306例临床血清采用ELISA方法检测HBVM和PreS1,荧光定量PCR法检测HBV DNA。结果：中HBeAg阳性146例,其中前S1阳性127例阳性率88.1%,HBeAg阴性共160例,其中前s72例阳性率45%,HBeAg阴性血清160例前s多数存在于抗HBe阳性血清中,阳性率40.6%。结论：PreS1比HBeAg更能较好反映病毒存在和复制状态.HBVM、PreS1和HBV DNA联合检测,更有利于乙肝病情监测和疗效评估。     

乙肝病毒外膜大蛋白检测的临床意义

目的:探讨乙肝病毒外膜大蛋白(HBV-LP)检测的临床意义.方法:随机选取203例乙肝患者血清,采用酶联免疫吸附试验(ELISA)检测HBV-LP、Pre-S1蛋白及HBV M,实时定量PCR方法检测HBV DNA.结果:LP的检出阳性率与HBV DNA的检出阳性率相关性具有统计学意义,且HBV DNA 拷贝数的对数值与HBV-LP 表达具有相关关系(r＝0.902);LP检出阳性率与Pre-S1蛋白检出阳性率差异具有统计学意义;不同HBV M模式检测LP检出阳性率差异不具有统计学意义,但26例HBeAg阴性患者血清LP检测为阳性.结论:血清中HBV-LP的含量与HBV DNA的拷贝数具有较好的相关性;血清中HBV-LP的检测是对HBV M检测的补充.     

Cytomegalovirus infection in patients with kidney diseases

Cytomegalovirus (CMV) has a prominent position as the cause of severe cytomegalovirus infection (CMVI) in immunocompromised persons, such as: patients with primary or secondary immunodeficiency (risk group), blood/hemoproducts recipients, especially recipients of tissue and organ transplants, which have implications for difficult providing of CMV-seronegative (safe) blood. However, CMV-disease is manifested in the minority of immunocompetent persons. Sera of patients from the Center for Hemodialysis of the Clinic of Nephrology of the Military Medical Academy (MMA) were tested on the presence of CMVI markers. All patients belonged to the risk group considering the chronicity of the main disease, receiving of the large quantities of blood and as the possible potential recipients of kidney transplants. Testing was performed by commercial serologic-enzyme-immune tests (Abott--USA, Behring--Germany) for CMV antibodies of IgG and IgM classes by which was determined serologic state of patients. Test results: out of 106 tested sera samples, 100 (99.33%) were CMV-seropositive (CMV-IgG), 27 (25.47%) were positive on recent CMVI (CMV-IgM), 99 (93.39%) received the blood previously, the majority of patients were with the diagnosis of primary glomerulopathy--68 (64.15%) and tubulointerstitial nephropathy--32 (30.18). Such results indicate the potentional possibility of reactivation of latent CMV and development of CMVI, and in transplant recipients--its rejection. It can occur if appropriate measures of prevention from CMVI are not timely undertaken, which includes the providing of CMV-seronegative-safe blood/hemoproducts, as well as the appropriate application of combined medicamentous therapy by chemical agents and immunomodulators-specific hyperimmune anti-CMV immunoglobulins.     

Automated quantitative evaluation of diseased and nondiseased renal transplants with MR renography

PURPOSE: To present a method of automated parametric quantification of dynamic MR enhancement curves of renal transplants and evaluate the disease-discriminating properties of the resulting MR renography (MRR) data. MATERIALS AND METHODS: This study included 27 patients with nondiseased renal transplants and eight patients with diseased renal transplants. The examination was repeated in 10 patients and the reproducibility of the enhancement parameters was estimated by analysis of variance (ANOVA). The disease-discriminating properties of the transplant volumes and enhancement parameters were tested with t-tests and logistic regression analysis. RESULTS: The enhancement parameters were reproducible. The mean medullary nephronal washout rate (lambda1) and cortical arterial blood volume (mu0) were lower in diseased renal transplants. The combination of these parameters was a strong predictor of renal transplant disease (area under ROC curve 0.98; 95% confidence interval 0.96-1.0). CONCLUSION: Automated parametric quantification of cortical and medullary enhancement is feasible and allows the accurate detection of nonsurgical disease in renal transplants by MRR.     

The role of interventional radiology in the management of kidney transplant complications

PURPOSE: To evaluate the role and the effectiveness of interventional radiology in the treatment of renal transplant complications. MATERIALS AND METHODS: From 1996 to 2004 a total of 288 kidney transplants from cadavers were performed in our Institute. The kidney was always collocated in iliac fossa by creating a vascular anastomosis with the external iliac artery and vein; in all cases the ureter was implanted into the recipient bladder. During the follow-up, 34 complications were observed. Twenty-seven complications in 25 patients (20 males and 5 females; age 35-65 years) were treated by a radiologic procedure: 9 renal artery stenosis and 1 native external iliac artery stenosis (by PTA), 5 ureteral obstructions (by nephrostomy and ureteral stenting), 8 ureteral leaks (by nephrostomy, in 2 cases associated to ureteral stenting) and 4 limphoceles (by percutaneous ultrasound-guided catheter drainage). RESULTS: Primary technical success was obtained in 20/27 cases (74%). Success was obtained with a second interventional procedure in 3/27 cases, 2 limphoceles and 1 ureteral fistula (secondary technical success: 85.2%), with a clinical final success in 23/27 cases (85.2%). We observed a peri-procedural complication rate of 3.7% (1 renal artery post-PTA dissection during a restenosis treatment). Four cases (1 renal arterial post-PTA dissection, 1 ureteral obstruction, 1 ureteral leak and 1 limphocele) needed a surgical correction (14.8%). CONCLUSIONS: Interventional radiology is the first therapeutic approach to treat renal transplant complications. It shows good technical and clinical results and a low complication rate. Surgery had to be considered only if minimally invasive procedures are infeasible or ineffective.     

原发性高血压病患者尿微量蛋白测定与管型蛋白质分析的临床意义

用ELISA法检测微量尿白蛋白(Alb)、IgG、IgA、IgM,用酶标抗体染色作尿管型蛋白成份分析,对123例原发性高血压患者进行了观察。结果提示,高血压患者尿Al1b、IgG、IgA、IgM显著升高,与疾病严重程度、血压高低、病程长短有密切关系。治疗后可降至正常。患者管型中不含IgG、IgA、IgM与Alb。两法联合应用,有助于区别肾性与原发性高血压,并可对原发性高血压的肾功能改变进行早期预测与监护。     

人巨细胞病毒糖蛋白B抗原表位在大肠杆菌中的表达

目的:利用大肠杆菌表达人巨细胞病毒(HCMV)糖蛋白B抗原表位AD1和AD2,对表达产物进行纯化,通过ELISA方法检测表达产物与感染HCMV的人血清之间的特异结合反应,探讨其用于临床检测的可行性. 方法:以HCMV病毒基因组为模板,PCR扩增AD1和AD2基因,构建重组表达载体pET32-NusA-AD1和pET32-NusA-AD2,在大肠杆菌BL21(DE3)pLysS中利用IPTG诱导表达,通过Ni柱亲和纯化获得NusA-AD1和NusA-AD2, ELISA方法检测NusA-AD1和NusA-AD2与感染HCMV的人血清之间的特异性结合反应.结果:可溶性表达并纯化得到融合抗原NusA-AD1和NusA-AD2.在8份已确定为HCMV IgG阳性人血清标本中,融合抗原NusA-AD1能够与5份血清发生反应,融合抗原NusA-AD2能够与6份血清反应.结论:NusA-AD1和NusA-AD2能够部分检测HCMV感染的人血清.     

SARS 25例临床诊断患者血浆特异性抗体和病毒的动态检测

目的 动态检测SARS患者血浆特异性IgM和IgG以及SARS冠状病毒(SARS-CoV)的变化规律。方法 用酶联吸附免疫法对25例145份SARS临床诊断患者血浆特异性IgM和IgG进行定性检测,用RT-PCR对其中114份血浆进行SARS-CoV定性检测。结果 IgM和IgG抗体的阳性样本检出率分别为49．0％(71／145)和54．5％(79／145),阳性患者检出率均为84．0％(21／25),两种抗体大多在发病第2～4周产生,前5周检出率相近,均呈上升趋势,以后IgM抗体检出率开始下降,IgG抗体检出率则继续上升。血浆SAPS-CoV阳性样本检出率为15．8％(18／114),阳性患者检出率为40．0％(10／25),多为发病后4周内采集的样本,抗体检测阴性或初次阳性。结论 血浆特异性抗体以及SARS-CoV检测可作为SARS的确诊依据;抗体产生后大多数病人血浆病毒很快转阴,但有个别病人在抗体产生2～3周后仍能在血浆中检测到病毒序列。     

Strahlentherapie im Rahmen der allogenen Nierentransplantation: eine Indikation zur lokalen Bestrahlung?

PURPOSE: In the past decades the indications for local graft irradiation (LGI) in acute renal transplant rejection have been limited and considered unfavorably. Despite major advantages in maintenance immunosuppression and management of acute allograft rejection a minority of patients remains with drug resistant transplant rejection. This subgroup of patients may benefit from LGI. PATIENTS AND METHODS: Between 1979 and 1990, eight patients with biopsy-proven acute renal allograft rejection and failure of all other immunosuppressive measures (corticosteroids, ATG, ALG or OKT3) were treated with LGI. Retrospective analysis was conducted for this control group. Radiotherapy was performed with Co-60 up to a median total dose of 6.0 Gy (single doses: 1.5-2.0 Gy). Six of eight patients were dialysis dependent prior to irradiation. In addition a literature review was performed including most important textbooks, electronic databases (Medline, Embase, Science Citations Index), and the internet. RESULTS: Two of eight patients experienced a clinical reversal of rejection and an improvement of renal function: serum creatinine decreased significantly. One patient remained free of dialysis with a functioning graft, the other had a recurrent rejection 2 months later and became dialysis dependent. The literature review showed, that adjuvant LGI has no advantage over conventional immunosuppression. However, in case of a drug refractory allograft rejection LGI restores long-term stable organ function in 13-60% of cases. CONCLUSION: The value of LGI of organ transplants, like renal allografts, is still not clearly defined. As a rescue measure in drug refractory allograft rejection special patients may clinically benefit, when a transplant nephrectomy can be avoided. Further prospective clinical trials are needed for a better assessment of LGI in organ transplantation.     

肾移植后巨细胞病毒感染的诊断与治疗

研究肾移植术后巨细胞病毒(CMV)感染的诊断及预防性使用磷甲酸钠(PFA)治疗的疗效与毒副作用。采用ELISA CMV特异性抗体IgG、IgM方法，针对肾移植术后74例CMV活动期感染(34例有明显临床症状)和25例静止期患者使用PFA，另选l0例静止期病例不予处理作为对照。结果显示，无症状的活动期病例PFA平均使用12．5天后血清学结果转阴；有临床症状的34例中，应用15天后32例显效，另2例治疗1个月后血清学结果转阴，随访1年无复发；静止期治疗组用药后观察1年，发现2例血清IgM转阳，但无临床症状；对照组l0例中有3例分别于随访l、2、3个月时发生CMV活动性感染。研究表明，PFA可快速治疗CMV感染，疗效确切而显著，随访期内复发少见；静止期患者使用PFA与否，随访期内血清学的检查结果差异显著；PFA对移植肾功能无明显损害，不干扰血钙及环孢素A的代谢，是一安全而有效的治疗CMV感染的药物。