人巨细胞病毒PP65抗原检测在中枢神经系统感染性疾病中的应用

人巨细胞病毒（HCMV）可侵袭中枢神经系统，引起相应的感染性疾病及并发症甚至死亡。HCMV的低基质磷酸蛋白65（HCMV-PP65）抗原是HCMV感染表达的早期抗原，用单克隆抗体混合物C10／C11检测HCMV-PP65抗原，是监测HCMV在疾病中活动性感染的标准方法之一。本研究对25例中枢神经系统感染患者进行了HCMV-PP65抗原检测，现报告如下。     

母乳及婴儿尿液人巨细胞病毒DNA检测在婴儿人巨细胞病毒感染中的应用

目的:定量检测疑似人巨细胞病毒(HCMV)感染婴儿尿液及对应母亲乳汁中HCMV DNA,评估两者在诊断HCMV感染中的价值,并比较两者浓度之间的关系。方法:选取51例疑似HCMV感染的婴儿,收集其新鲜尿液及对应的母亲乳汁,分别用荧光定量聚合酶链式反应法(FQ-PCR)检测HCMV DNA。结果:51例疑似HCMV感染婴儿尿液中,有49例检测出HCMV DNA,阳性率为96.0%(49/51)。51份婴儿母亲乳汁中,有37份检测出HCMV DNA阳性,阳性率为72.5%(37/51)。对37例尿液与乳汁HCMV DNA均为阳性的患儿,分别对尿液与乳汁HCMV DNA浓度取对数,将两者进行相关性分析,结果显示两者之间存在一定的相关性,相关系数为0.332(P<0.05)。结论:婴儿尿液中HCMV DNA检测对于婴儿HCMV的感染具有重要的价值,同时对其母乳HCMV DNA检测,可以发现婴儿尿液和母乳HCMV DNA具有一定的相关性。     

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     

应用间接免疫荧光技术检测人巨细胞病毒-PP65抗原的临床意义

目的：探讨人巨细胞病毒（HCMV）-PP65抗原检测的临床意义。方法：利用间接免疫荧光技术对本院56例住院患者外周血HCMV-PP65抗原进行检测,其中器官移植组35例（肾移植患者13例,肝移植患者14例,骨髓移植患者8例）,其他疾病组21例。设本院健康体检者20例为正常对照组。结果：56例标本中有25例检出HCMV-PP65抗原阳性,阳性率为45%。25例抗原阳性患者中,有临床症状的患者外周血中的HCMV-PP65抗原阳性细胞数明显高于无临床症状的HCMV-PP65抗原阳性的患者,两者比较差异有显著性（P〈0.01）。正常对照组无一例阳性。结论：HCMV-PP65抗原检测具有简便、敏感的特点,其结果与患者的临床症状相关,可以作为监测HCMV活动性感染的指标,为临床提供诊治依据。     

用巨细胞病毒阴性供血者的血清识别巨细胞病毒临床分离抗原

背景防止CMV经输血传播最有效的方法是使用血清反应阴性供血者的血制品。供血者的CMV筛查通常是建立在检测取自CMV实验室提取的AD169抗原的检出。近来有迹象表明大约高达20％的CMV阴性供血者实际上经PCR分析为CMV-DNA阳性。研究设计及方法在本研究中，作者检测了CMV血清反应阴性血清，CMV血清反应阳性血清和CMV     

不同标本巨细胞病毒检测在诊断小儿人巨细胞病毒感染中的应用

目的：探讨尿液、淋巴细胞和母乳的 HCMV-DNA 检测以及血清 HCMV-IgM 检测在小儿人巨细胞病毒（HCMV）感染诊断中的意义。方法用实时荧光定量 PCR检测189例疑似HCMV感染患儿尿液、淋巴细胞和母乳的HCMV-DNA ,用化学发光法检测其血清HCMV-IgM ,并分年龄分性别对其阳性率进行分析比较。结果新生儿组母乳HCMV-DNA阳性检出率（60.3％）显著高于尿液（5.2％, P〈0.001）和淋巴细胞（12.1％,P〈0.001）,也显著高于血清 HCMV-IgM 阳性率（3.4％,P〈0.001）;婴儿组母乳（48.1％）与尿液（55.0％）HCMV-DNA 阳性率差异无统计学意义（P〉0.05）,但两者均显著高于淋巴细胞 HCMV-DNA 阳性率（22.1％, P〈0.001）和血清 HCMV-IgM 阳性率（24.4％, P〈0.001）;婴儿组的尿液 HCMV-DNA、血清 HCMV-IgM 阳性率均显著高于新生儿组（ P〈0.01）;两组各指标男女阳性率差异均无统计学意义（ P〉0.05）;母乳 HCMV-DNA 阳性率与尿液、淋巴细胞 HCMV-DNA 阳性率及血清 HCMV-IgM 阳性率均呈显著正相关（r ＝0.630,P ＝0.000;r ＝0.413,P ＝0.000;r ＝0.341,P ＝0.000）。结论联合尿液、淋巴细胞HCMV-DNA 和血清HCMV-IgM 检测有助于患儿HCMV 感染的辅助诊断;在无法联合检测的条件下,婴儿首选尿液 HCMV-DNA 检测可获得较高检出率;母乳 HCMV-DNA检测并正确喂养有助于预防HCMV感染。     

异基因造血干细胞移植后巨细胞病毒抗原血症检测指导巨细胞病毒感染的防治

人巨细胞病毒感染及巨细胞病毒疾病是异基因造血干细胞移植后的常见并发症,巨细胞病毒性间质性肺炎是移植后巨细胞病毒疾病的主要类型,如未能及时检出和救治则病死率极高,为探讨采用免疫组化法测定人巨细胞病毒的方法学,指导临床治疗,我们采用免疫组化法对45例（慢性粒细胞白血病23例,急性髓性白血病7例,急性淋巴细胞白血病6例,骨髓增生异常综合征4例,非何杰金氏淋巴瘤3例,再生障碍性贫血2例）进行造血干细胞移植后巨细胞病毒抗原血病的检测,并与巨细胞病毒血清学检测进行了对比,45例中38例患进行了异基因外周血干细胞移植。2例骨髓移植,5例外周血与骨髓共移植,移植前,对供/受均进行巨细胞病毒抗体检查;移植后,受采用本方法进行抗原检查,结果表明,有25例患发生巨细胞病毒相关性间质性肺炎移植后随访时间为6-28个月（平均时间为18个月）,根据巨细胞病毒检测结果进行预防性治疗的患预后较未进行预防性治疗的患有明显差异。预防组巨细胞病毒相关的死亡率为1/29,未预防组为12/16,提示采用免疫组化法对异基因造血干细胞移植患进行定期随访监测可更早期、快速的检出巨细胞病毒感染,对患预后有重要意义。     

人巨细胞病毒抗体筛查预防输血性人巨细胞病毒感染的Meta分析

目的明确输注人巨细胞病毒(CMV)血液抗体筛查阴性血在减少免疫低下人群输血性CMV感染的作用,为CMV抗体血液筛查的实施提供依据。方法计算机检索PubMed、MEDLINE、Ovid、ProQuest、EBSCO、Cochrane Library、EMbase、CNKI、VIP、CBM和WanFang数据库,检索比较输入CMV血液抗体筛查阴性血与输入未经筛查且为非去白细胞处理血对输血性CMV感染率差异的文献,同时辅以手检纳入文献的参考文献。按照纳入排除标准进行筛选、资料提取和质量评价后,使用Rev Man5.1软件进行Meta分析。结果共纳入7项研究,430例患者。Meta分析结果显示输入经CMV血液抗体筛查阴性血与输入未经筛查且为非去白细胞处理血后,受血者的输血性CMV感染率差异具有统计学意义(OR=0.07,95%CI:0.03~0.18,P0.01)。结论进行CMV抗体血液筛查能有效减少免疫低下受血者输血性CMV感染率,特别对器官移植和新生儿输血患者作用明显。     

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ＷＢＣ，在ＣＭＶ病者为２０．５±１７．８／５万ＷＢＣ。同时检测１０名健康人，结果均为阴性。结论该法具有快速、简便、成本低等优点，且能达到定量检测的目的，可用于临床对ＣＭＶ感染的早期快速诊断。     

人巨细胞病毒糖蛋白gp52的原核表达及初步应用

目的探讨以表达人巨细胞病毒（ＨＣＭＶ）中抗原性较强的蛋白片断代替全病毒抗原建立ＨＣＭＶＩｇＭ检测试剂盒的可行性。方法以聚合酶链反应扩增ＨＣＭＶ聚合酶辅助糖蛋白（ｇｐ５２）的有效抗原基因序列，导入带有高效转录启动子ｐＴｒｃ和６个串联组氨酸的原核表达载体ｐＴｒｃＨｉｓ，在大肠杆菌中表达，经金属鏊和亲和层析（ＩＭＡＣ）法纯化后，用酶联免疫吸附法检测新生儿血清，并与全病毒抗原检测进行比较。结果重组ｇｐ５２占大肠杆菌总蛋白的４０％，ＩＭＡＣ纯化后纯度达９４．５％。在４３份疑似ＨＣＭＶ感染的新生儿血清中阳性检出率为５１．２％，而用全病毒抗原检测阳性率只有２５．９％。结论ＨＣＭＶｇｐ５２蛋白具有良好的抗原性，可用于建立新一代的检测试剂盒。     

A cohort study of the universal neonatal urine screening for congenital cytomegalovirus infection

ObjectivesThis prospective cohort study aimed to evaluate the efficacy of the universal neonatal urine screening, followed by diagnosis, workup and antiviral therapy for symptomatic congenital cytomegalovirus (CMV) infection to reduce neurological impairments and sequelae.MethodsNeonates born in three facilities underwent the universal urine screening of PCR analyses for CMV-DNA. Neonates with symptomatic congenital CMV infection (cCMV) received oral valganciclovir (VGCV) of 32 mg/kg/day for six weeks or six months, and were evaluated for neurological outcomes including developmental quotient (DQ) and hearing function at around 18 months of corrected age.ResultscCMV was diagnosed in 56 (0.48%) of 11,736 neonates, consisting of 23 neonates with symptomatic and 33 with asymptomatic cCMV. The incidence of cCMV in the general perinatal medical center (0.69%) was higher than that in the primary maternity hospital (0.23%, p<0.01%). Twenty of the 23 infants with symptomatic cCMV received VGCV therapy, and 19 underwent neurological assessment. Eight neonates (42%) had severe sequelae of DQ < 70, bilateral hearing dysfunction, and/or epilepsy. Four neonates (21%) had mild sequelae of DQ 70–79 or unilateral hearing dysfunction only, and seven (37%) showed normal development without any impairment.ConclusionsThis study on a large scale demonstrated that a series of universal neonatal urine screening, diagnosis, workup, and VGCV therapy for neonates with symptomatic cCMV may decrease neurological impairments, because 58% of the treated infants had normal development or mild sequelae. The universal urine screening likely identifies subclinical symptomatic cCMV. Mothers with fetuses of cCMV seem to be selectively transferred to perinatal medical centers before deliveries.     

早产儿先天性巨细胞病毒感染特点及危险因素分析

目的调查早产儿先天性巨细胞病毒(CMV)感染状况,分析其特点及危险因素。方法选择住院的早产儿,分别采用荧光定量PCR(FQ-PCR)法和ELISA法检测脐血血清CMV IgM与DNA。同时记录新生儿和母亲的人口学信息,采用二元多因素logistic回归分析早产儿先天性CMV感染相关影响因素。结果共纳入1315例早产儿,血清CMV IgM和(或)CMV DNA阳性者占1.98%(26/1315),CMV IgM阳性者占1.44%(19/1315),血清CMV DNA阳性者占1.14%(15/1315),CMV IgM与CMV DNA均为阳性者占0.61%(8/1315)。早产儿先天性CMV感染症状较为轻微。母亲年龄<25岁、初次妊娠、孕期胎膜早破是早产儿先天性CMV感染的危险因素(P均<0.05)。结论早产儿先天性CMV感染发生率较高,以无症状感染为主。提高年轻育龄妇女对CMV的知晓率、加强早产儿先天性CMV感染的管理是很有必要的。     

Clinical factor associated with congenital cytomegalovirus infection in pregnant women with non-primary infection

The aim of this nested case-control study was to evaluate clinical factors associated with the occurrence of congenital cytomegalovirus (CMV) infection in pregnant women with non-primary CMV infection. In a cohort study of CMV screening for 2193 pregnant women and their newborns, seven newborns with congenital CMV infection were identified among 1287 pregnant women with non-primary CMV infection that was defined as negative IgM and positive IgG with IgG avidity index >45%. In the 1287 women with non-primary CMV infection, clinical findings and complications were compared between pregnancies with and without congenital CMV infection. Clinical factors associated with the occurrence of congenital CMV infection were evaluated. The birth weight of newborns with congenital CMV infection was less than that of newborns without congenital infection (p < 0.05). Univariate logistic regression analyses demonstrated that threatened premature delivery (OR 10.6, 95%CI 2.0–55.0; p < 0.01) and multiple pregnancy (OR 7.1, 95%CI 1.4–37.4; p < 0.05) were associated with congenital infection. Multivariable logistic regression analyses demonstrated that threatened premature delivery (OR 8.4, 95%CI 1.5–48.1; p < 0.05) was a single risk factor for congenital CMV infection in pregnant women with non-primary CMV infection. This study revealed for the first time that threatened premature delivery was associated with the occurrence of congenital CMV infection in pregnant women with non-primary CMV infection, the pathophysiology of which may be closely associated with CMV reactivation during pregnancy.     

巨细胞病毒先天性感染与低出生体重儿关系的研究

目的：探讨巨细胞病毒（ＣＭＶ）的先天性感染对新生儿胎龄、体重的影响。方法：对１８６例早产适于胎龄儿、１４２例小于胎龄儿（ＳＧＡ）和９５例足月儿与其母配对,用酶联免疫吸附法和免疫组织化学法分别检测抗ＣＭＶ特异性ＩｇＭ抗体及巨细胞病毒抗原（ＣＭＶＡｇ）。结果：ＣＭＶ先天性感经ＳＣＡ组为３１．７％,明显高于早产适于胎龄儿的８．１％和足月儿对照组的５．２％,结论：小儿胎龄儿的产生可能与ＣＭＶ的先天性感染有     

Efficacy of prolonged valganciclovir therapy for congenital cytomegalovirus infection

We report prolonged valganciclovir (VGCV) treatment of a symptomatic cytomegalovirus infection case. Automated auditory brainstem evoked response performed at 5 days of age revealed severe hearing impairment. Cranial magnetic resonance (MR) imaging at 11 days of age showed abnormal findings. At 5 weeks of age, VGCV was started. The viral load in blood cells, plasma, and urine decreased during the 6-week treatment. Because of improvement of hearing level and no adverse effects, VGCV was restarted for an additional 6 weeks. Neither the patient’s hearing impairment nor results of cranial MR imaging have become worse in 6 months. It is crucial to gather information from as many cases as possible treated with VGCV to establish a standard protocol for VGCV treatment.     

Optimum treatment of congenital cytomegalovirus infection

Congenital cytomegalovirus infection affects 0.7% of live births and is the leading cause of congenital neurological handicaps of infectious origin. However, systematic screening of this infection has not been implemented in pregnancy or at birth in any country. This apparent paradox has been justified by the unavailability of an efficient vaccine and by the scarcity of data available on the treatment of congenital CMV. However, in the last decade interesting new data on the management of this congenital infection has emerged including new results on both neonatal and postnatal treatments. This review provides an update on the potential benefits of antiviral treatment and on passive immunisation both in the neonatal and the antenatal periods. These suggest a benefit to a proactive approach for neonatal and prenatal congenital infections.     

应用半巢式核酸荧光定量技术检测肾移植术后人巨细胞病毒感染

目的：建立一种新的用于肾移植术后人巨细胞病毒（ＨＣＭＶ）感染的快速定量诊断方法,方法：采用半巢式酸荧光寂量技术（ＡｍｐｌｉＳｅｎｓｏｒ－ＰＣＲ）检测已知含量的ＨＣＭＶ－Ａ在６９标准病毒株,无关病毒,无关细菌及临床血标本的ＨＣＭＶ－ＤＮＡ和质粒标准品,以确定该方法的敏感性,特异性和定量的准确性。结果：对已知含量的标准病毒株的定量检测结果与其已知含量基本一致;无关病毒包括单纯疱疹病毒１型（ＨＳＶ－１）     

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