慢性肾盂肾炎患者人巨细胞病毒感染的分析

慢性肾盂肾炎是尿路感染的常见病 ,除细菌学病因外 ,有报道与人巨细胞病毒 (ＨＣＭＶ)感染有关[1 ,2 ] 。为进一步探讨慢性肾盂肾炎同ＨＣＭＶ感染的关系 ,我们同时检测慢性肾盂肾炎患者尿中ＨＣＭＶ抗原和ＨＣＭＶＤＮＡ及血清ＨＣＭＶ抗体(ＩｇＧ型 ) ,现报道如下。材料与方法一、对象患者组 :临床确诊的慢性肾盂肾炎患者 95例 ,其中男 31例、女 64例 ,年龄 2 9～ 44岁 ;正常对照组 :83例 ,其中男 2 8例、女 5 5例 ,年龄 2 0～ 47岁 ,经体检和询问均无慢性肾盂肾炎病史及临床症状。二、试剂和方法1 .标本的收集和预处理　尿液标本 :取两组…     

人类巨细胞病毒感染与肺癌关系的研究

本文检测了５３例肺癌，５７例肺非癌患者和１００例自然人群血清特异性巨细胞病毒的ＩｇＭ和ＩｇＡ抗体，并检测了其３２例肺癌患者支气管－肺泡灌洗液中的ＣＭＶ－ＤＮＡ序列，肺癌患者血清中ＣＭＶ抗体阳性共２４例（４５．３％），明显高于肺非癌患者和自然人群，３２例肺癌患者ＢＡＬＦ中的ＣＭＶ－ＤＮＡ序列检出８例（２５．０％），与ＣＭＶ－ＩｇＭ抗体阳性结果（２８．１％）相符，从肺癌患者血清和ＢＡＬＦ中分别检测到Ｃ     

早期快速诊断小儿柯萨奇B组病毒感染的研究

早期快速诊断小儿柯萨奇Ｂ组病毒（ＣＢＶ）感染。方法采用酶联免疫吸附（ＥＬＩＳＡ）方法，检测１１２例疑似ＣＢＶ感染的新生儿血清中ＣＢＶＩｇＭ抗体。同时将ＥＬＩＳＡ方法与病毒分离法进行比较。结果１１２例疑似ＣＢＶ感染新生儿血清中ＣＢＶＩｇＭ阳性６８例，阳性率６０７１％；ＥＬＩＳＡ方法与病毒分离法比较阳性符合率９２６５％，阴性符合率９７７３％。结论ＥＬＩＳＡ方法适于早期快速诊断小儿ＣＢＶ的感染     

丙型肝炎IgM抗体检测及其临床意义

为探讨丙型肝炎ＩｇＭ抗体的临床意义，建立了间接酶联免疫吸附法（ＥＬＩＳＡ），该方法重复性好、特异性强、操作简便。检测２２例急性丙型肝炎和５０例慢性丙型肝炎患者的血清，结果单独使用核心区抗原检测ＩｇＭ抗体的阳性率大于用核心区和非结构区３抗原共同检测。抗－丙型肝炎病毒（ＨＣＶ）ＩｇＭ在急性丙型肝炎患者中检出率为８６％，慢性丙型肝炎中检出率为６６％，但急性组中５０％患者ＩｇＭ出现（约于输血后７．０±３．７周即可在血清中测出）早于ＩｇＧ，而慢性组中无一例如此，两组间差异有非常显著意义（Ｐ＜０．００５）。急性组血清丙氨酸转氨酶显著升高患者中８０％可测出ＩｇＭ；慢性组中６７％可测出ＩｇＭ。ＨＣＶＩｇＭ对急性丙型肝炎的诊断是一个有价值的指标，同时与疾病的活动性有一定相关性。     

Raji细胞免疫酶抑制法检测血清抗HLA－DR抗体

本研究利用Ｒａｊｉ细胞表面具有表达组织相容性抗原ＤＲ（ＨＬＡ一ＤＲ）特点，以及被检血清中抗ＨＬＡ一ＤＲ抗体与抗人ＨＬＡ一ＤＲ单克隆抗体（ＭｃＡｂ）竞争结合Ｒａｊｉ细胞表面ＨＬＡ一ＤＲ抗原的特征，建立检测血清抗ＨＬＡ一ＤＲ抗体的Ｒａｊｉ细胞免疫酶抑制法。对７２例系统性红斑狼疮（ＳＬＥ）患者和１１３例健康者血清检测结果表明：ＳＬＥ患者和健康各血清中抗ＨＬＡ一ＤＲ抗体阳性率分别为３９．０％和１．８％。此法特异性强、重复性好（阳性血清ＣＶ为４．７％，阴性血清ＣＶ为０％），抗体涌度范围１：５～１：４０。     

逆转录套式ＰＣＲ检测丙型肝炎病毒感染者ＨＣＶ ＲＮＡ

目的　探讨人血清和外周血单个核细胞（ＰＢＭＣ）中丙型肝炎病毒（ＨＣＶ）ＲＮＡ与血清ＩｇＧ和ＩｇＭ抗体联合检测对丙型肝炎的诊断价值。方法对以ＥＬＩＳＡ方法检测所得的４６例ＨＣＶ—ＩｇＧ阳性的标本用逆转录套式聚合酶锭反应（ＲＴ—ＮｅｓｔｅｄＰＣＲ）检测血清和ＰＢＭＣ中ＨＣＶＲＮＡ，同时用ＥＬＩＳＡ方法检测血清中ＨＣＶ—ＩｇＭ。结果血清中有１７例ＨＣＶ　ＲＮＡ阳性和６例ＨＣＶ—ＩｇＭ阳性，而ＰＢＭＣ中有１９例ＨＣＶＲＮＡ阳性。结论血清中ＨＣＶ—ＩｇＭ可作为丙型肝炎的初筛诊断，ＨＣＶ—ＩｇＭ可作为ＨＣＶ近期感染的指标，但二者均不能准确反映病毒在体内的存在情况，而套式ＰＣＲ方法检测ＨＣＶＲＮＡ可直接反映ＨＣＶ在体内的活动情况，其中ＰＢＭＣ中检测ＨＣＶＲＮＡ的阳性率高于血清，提示ＨＣＶ可能在ＰＢＭＣ中潜伏乃至复制。     

白细胞中巨细胞病毒抗原检测及在肾移植的临床应用

目的 建立一种新的快速诊断巨细胞病毒（ＣＭＶ）感染的方法并初步用于肾移植受者，方法 采用抗ＣＭＶ抗原的单克隆抗体，运用免疫组化链菌素－生物素标记法对外周血白细胞核中ＣＭＶ抗原（即刻早期抗原和早期抗原）进行染色，诊断ＣＭＶ感染。结果 共检测肾移植受者７２例，ＣＭＶ活动性感染率为４７％（３４／７２），ＣＭＶ病发病为１８％（１３／７２），ＣＭＶ抗原阳性细胞数在ＣＭＶ活动性感染者中平均为１１．５／５万ＷＢ     

血清丙型肝炎病毒核心抗原、ＮＳ３抗原的检测及临床意义

目的　为检测病人血清中丙型肝炎病毒（ＨＣＶ）核心抗原、ＮＳ３抗原井探讨其临床意义。方法　用基因工程表达的ＨＣＶ核心蛋白、ＮＳ３抗原制各其单克隆抗体，建立检测病人血清中ＨＣＶ核心抗原、ＮＳ３抗原的ＥＬＩＳＡ法。结果　６０例慢性丙型肝炎患；２５０例杭—ＨＣＶ—１８Ｇ阳性；３０例抗—ＨＣＶ—ＩｇＧ、ＩｇＭ均阳性；２００例透析病人；２０例白血病病人；１５０例普通病人中ＨＣＶ核心抗原、ＮＳ３抗原阳性检出率分别为：６．６％、８．４％、１６．７％、４％、５％、２％和１３．３％、１０．４％、２３．３％、３．０％、５．０％、２．６％。结论ＨＣＶ核心抗原、ＮＳ３抗原的检测，可作为现有检测方法的补充，能更好地反映患体内ＨＣＶ的活动情况。     

应用双抗原夹心法检测抗－HCV抗体

目的建立检测抗－ＨＣＶ抗体的双抗原夹心酶联免疫吸附测定（ＥＬＩＳＡ）方法。方法用基因工程手段，在大肠杆菌中表达ＨＣＶ抗原表位与大肠杆菌噬菌体ＭＳ２ＤＮＡ聚合酶（ＭＳ２－Ｐｏｌ）的融合蛋白，纯化后作为包被抗原，可溶性表达的ＨＣＶ抗原表位与霍乱毒素Ｂ亚基（ＣＴＢ）的融合蛋白经辣根过氧化物酶标记后作为酶结合物，建立双抗原夹心ＥＬＩＳＡ。结果ＭＳ２－Ｐｏｌ融合蛋白在大肠杆菌中得到高效表达并进行了纯化，所建立的双抗原夹心ＥＬＩＳＡ系统用于检测１４份ＨＣＶ阳性血清样品，抗－ＨＣＶ抗体检出率１００％；检测２１份ＨＣＶ阴性血清，没有假阳性结果。ＣＴＢ抗体不干扰反应。结论双抗原夹心法与采用二抗的ＥＬＩＳＡ系统具有相近的灵敏度和更好的特异性。可以避免由于二抗干扰带来的非特异性。     

荧光定量PCR诊断HCMV病毒感染的阈值建立

人巨细胞病毒（ＨＣＭＶ）在我国１０岁以下儿童的感染率（抗ＨＣＭＶ－－ＩｇＧ阳性率）超过９５％。当机体抵抗力低下时，ＨＣＭＶ可引起多种组织的感染甚至危及生命。多聚酶链反应可作为ＨＣＭＶ感染早期快速的检测手段，但由于该病毒潜伏感染的特点（多为肾脏），正常健康人的尿液内亦有少量的ＨＣＭＶ，一般定性ＰＣＲ可能存在一定的假阳性。因此我们采用荧光标记探针，以Ｔａｐｍａｎ技术对尿液中ＨＣＭＶ基因进行定量检测，观     

Heat-shock treatment in a rapid assay for cytomegalovirus detection in clinical samples

A mild heat-shock treatment of cells enhances cytomegalovirus (CMV) infection and shortens its eclipse period. This provided the basis for the development of a rapid assay to detect CMV in clinical samples.Urine specimens were inoculated in heat-shocked cells and then CMV-infected cells were visualized at various hours after infection with a monoclonal antibody directed against a CMV-induced immediate-early protein, using an immunoalkaline phosphatase staining. Out of 104 urine specimens examined, 13 proved positive and the assay we describe was able to yield positive results in a single day.     

柯萨奇病毒B RNA与中和抗体检测在病毒性心肌炎诊断中的意义

目的 探讨多重聚合酶链反应(PCR)及微量中和抗体法诊断病毒性心肌炎(MC)的价值。方法 用微量法和逆转录PCR法，检测100例诊断为急／慢性期MC患者和37名正常人中的柯萨奇B病毒核糖核酸(CVB—RNA)与中和抗体水平，并进行比较性研究。结果 正常人CVB—RNA及中和抗体均为阴性。在100例MC患者中，CVB—RNA阳性346例(34．6％)，其中急性期245例占70．8％，慢性期101例仅占29．2％。而中和抗体检测阳性及阴性各500例(均为50％)。在654例CVB—MA阴性患者中(65．4％)，中和抗体阳性者369例(56．4％)，其中慢性期255例占69．1％。急性期114例占30．9％。表明急性期CVB—RNA阳性居多，慢性期中和抗体阳性居多。结论 在MC的不同阶段，CVB—RNA和中和抗体的阳性率有所不同，同时进行2项检测可提高诊断率。     

Relationship between seropositivity of husbands and primary cytomegalovirus infection during pregnancy

The role of the sexual transmission of human cytomegalovirus (CMV) as a cause of congenital infection was investigated. Serum samples were collected from 756 pregnant women at 10 to 12 weeks of gestation and at 32 to 36 weeks of gestation. Serum samples were also obtained from the husbands of women who seroconverted and women who were seronegative during pregnancy. Commercially available enzyme immunoassay kits were used to detect serum IgG, IgM, and IgA antibodies against CMV. CMV from neonatal urinary specimens was isolated according to a standard tissue culture technique, using MRC-5 cells. At 10 to 12 weeks of gestation, 634 of the 756 pregnant women (83.9%) had IgG antibody to CMV. At 32 to 36 weeks of gestation, 642 of the 756 women (84.9%) had IgG antibody to CMV. A meaningful rise of serum IgG-antibody titer (seroconversion) occurred in 8 women (1.1%). CMV was isolated from the urine of an infant born to a seroconverted woman within a week after birth. The prevalence of IgG antibody to CMV was significantly higher in the husbands of women who seroconverted during pregnancy than in the husbands of the women who were seronegative during pregnancy (P < 0.01). Understanding the epidemiology of CMV is a key element in the development of strategies for the prevention of infection. The transmission of CMV by sexual contact may be important in the pathogenesis of congenital infection. Entirely new approaches to the prevention and treatment of congenital CMV infection are necessary, including antiviral interventions and the development of a vaccine strategy. Received: December 14, 1999 / Accepted: April 10, 2000     

尿巨细胞病毒-DNA载量与血清巨细胞病毒-IgM检测在新生儿巨细胞感染中的价值

目的 以实时荧光定量PCR法（FQ-PCR）检测尿液中巨细胞病毒载量,以化学发光法检测血清中CMV-IgM,探讨两种方法 联合应用在新生儿巨细胞病毒（CMV）感染中的临床价值.方法 144例疑似CMV感染新生儿的血清用化学发光检测CMV-IgM,尿液用FQ-PCR检测CMV-DNA载量,比较抗病毒治疗前、后两种检测结果 的变化.结果 两种方法 单独检测的阳性率分别为55.56%和66.67%,两者比较,差异无统计学意义（χ2=3.20,P＞0.05）,两种方法 联合检测的阳性符合率为76.39%（110/144）,明显高于两方法 单独检测阳性率,差异均有统计学意义（χ2分别=4.47、5.21,P均＜0.05）.抗病毒治疗后血CMV-IgM和尿CMV-DNA两种检测结果 均较治疗前明显下降,差异均有统计学意义（t分别=7.45、9.31,P均＜0.05）.结论 尿液CMV-DNA病毒载量与化学发光法血清CMV-IgM联合检测,可以明显提高新生儿巨细胞病毒感染检测的阳性率.     

婴儿肝组织巨细胞病毒抗原和病理检测

目的 :探讨婴儿巨细胞病毒 (CMV)感染患儿肝组织 CMV的感染及其病理改变。方法 :用免疫组化方法检测肝组织内 CMV前早期抗原和早期抗原 ,并在光学显微镜下观察肝组织病理改变。结果 :肝组织内检出 CMV- IEA2 0例和 CMV- EA均阳性 7例 ,阳性细胞多较集中分布 ,主要感染细胞为肝细胞、血管内皮细胞、胆管上皮细胞和炎性浸润细胞 ;肝组织内均有不同程度肝细胞肿胀变性或空泡形成 ,以及炎性细胞浸润 ,部分病例可见肝细胞内淤胆、汇管区纤维组织增生及肝细胞坏死 ;胆道畸形 10例肝细胞内淤胆。结论 :根据清抗 CMV- Ig M阳性和临床表现诊断为 CMV肝炎与肝组织 CMV抗原检测结果基本相符 ,CMV抗原阳性肝细胞病理改变与 CMV抗原阴性者无明显差异。     

Association of quantitative cytomegalovirus antigenemia with symptomatic infection in solid organ transplant patients

A prospective virologic follow-up of solid organ transplant patients was designed to determine the usefulness of antigenemia and viremia as virologic markers for the diagnosis of cytomegalovirus (CMV) infections, and also for monitoring CMV disease and therapy control. A total of 629 blood samples from 127 patients (60 liver, 47 kidney, and 20 heart transplant recipients) were studied by tube and shell vial cultures, and by antigenemia assay. This later was carried out by an indirect immunofluorescent assay method on formalin-fixed cytospin slides containing 2 × 10⁵ leukocytes, using a monoclonal antibody directed against the CMV pp65 antigen. CMV was detected by at least one of the three methods in 238 specimens (37.8%) from a total of 63 patients. The antigenemia assay was positive in 215 (90.3% of positive samples). A total of 94 samples were detected only by this marker, which occurred either in samples with low positive counts (70.2% with antigenemia counts <10 positive cells/10⁵ leukocytes) or in specimens from treated patients. There were 30 episodes of CMV disease in 23 patients. Antigenemia was positive in all these episodes, 27 of them with counts >20 positive cells/10⁵ leukocytes. With this cut-off, positive and negative predictive values for symptomatic CMV infection were 100% and 97.2%, respectively. The antigenemia assay is a rapid, sensitive, specific, and early marker of CMV infection in transplantees. Cultures became negative with antiviral therapy while remaining antigenemia detectable. There was an association between highest quantitative antigenemia test results and clinical symptoms in our patients. In its quantitative version, the assay is useful to detect symptomatic infection and appears to be a helpful tool in managing patients at risk and in guiding antiviral therapy.     

Urinary HIV-1 antibody patterns by western blot assay.

Diagnosis of human immunodeficiency virus infection (HIV-1) is normally carried out by serum testing for HIV-1 antibody. Recently, antibody testing in other body fluids such as saliva and urine have been attempted. In this study, we examined HIV-1 antibody patterns in urine by Western blot assay as compared to that found in serum. Out of 44 sero-positive samples by Western blot assay we found 43 to be HIV-1 antibody positive in the urine, whereas all 40 sero-negative samples were negative in urine. Thus the sensitivity of urine testing was 97.7% with 100% specificity when compared to serum testing by the Western blot assay. In the analysis of the antibody pattern in urine, we found 6.8% of p17, 68% of p24, and 47.7% of p39 in the core proteins; 72.7% of p31, 61.4% of p51, and 68.2% of p66 in the polymerase; and 63.6% of gp41, 75% of gp120, and 97.7% of gp160 in the envelope proteins. The data obtained supports the selection of the HIV-1 antigen subtype-E to develop a home test kit using urine. Urine testing for HIV-1 antibody is convenient, non-invasive, safe, and easily performed at home. However, if the urine is positive, the confirmation test on serum is needed.     

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

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。     

Reduction of postperfusion cytomegalovirus-infections following the use of leukocyte depleted blood

After extracorporeal perfusion some patients develop evidence of cytomegalovirus (CMV) infection. It is proposed that antigenically stimulated donor and recipient leukocytes may divide and activate latent CMV associated with leukocytes. To test this hypothesis, alternate patients were perfused with leukocyte-depleted or whole blood (controls). Four of six controls who were CMV antibody negative preoperatively seroconverted after perfusion. Virus was recovered from the blood of three and from the urine of two of these patients. One of four controls who were seropositive preoperatively developed a significant titer rise. Only one of eight patients perfused with leukocyte poor blond who were seronegative prior to surgery developed a viremia and became antibody positive; another manifested a viruria but remained antibody negative. These findings are consistent with the proposed hypothesis and suggest a means for reducing transfusion- associated CMV infections.     

The detection of cytomegalovirus DNA by the polymerase chain reaction in different biological materials in patients with organ allografts

Ninety-eight patients with organ allografts were examined using the polymerase chain reaction (PCR). Cytomegalovirus (CMV) DNA was not found in the blood of any of 67 blood donors. At least 85% patients with CMV DNA in the plasma developed CMV disease. CMV DNA was detected in leukocytes of 22 (40.7%) out of 54 patients with negative results of analysis for CMV DNA in the plasma; 16 (29.6%) of these 22 presented with clinical symptoms of CMV infection by the moment of investigation. Results of PCR analysis of blood, urine, and saliva specimens of 30 patients examined did not always coincide. DNA isolated from the urine can contain components inhibiting DNA polymerase. Blood leukocytes or whole blood are the most suitable for the diagnosis of active and symptomatic CMV-infection.