276例有异常妊娠史的妇女HCMV检测及分析

通过对1939例有正常妊娠史和异常妊娠(自然流产、过期流产、习惯性流产、葡萄胎、胚胎停止发育、死胎胎儿畸形等)史的生育妇女孕前进行巨细胞病毒(CMV)检测,结果发现有异常妊娠史的妇女人巨细胞病毒(HCMV)感染率(12.68％)比正常妊娠史的妇女HCMV感染率(8.0％)明显增高。因此,对生育妇女提供孕前期优生检测、对检测出HCMV阳性的高危人群实行积极治疗、跟踪随访、优生指导等综合管理,能明显地降低宫内感染和胎、婴儿传播的机会。有效的预防和降低因巨细胞病毒感染引起的病理性妊娠和残疾儿的出生,提高育龄妇女的生殖健康水平和人口出生质量。     

脐带血中巨细胞病毒DNA的检测

脐带血中巨细胞病毒ＤＮＡ的检测张吉旺，彭晓，朱俊芳人类巨细胞病毒（ＨＣＭＶ）感染可导致死胎、早产、畸形。新生儿巨细胞包涵体病和间质性肺炎等多种疾病，母婴垂直传播和输血是ＨＣＭＶ感染的主要途径。为探讨ＣＭＶ的先天感染率，我们对５６份正常胎儿脐带血单个核...     

TORCH与优生：Ⅰ．应用聚合酶链反应法辅助诊断巨细胞病毒性婴儿肝炎综合征

已知巨细胞病毒是婴儿肝炎综合征的常见病原之一,但临床生存诊断困难,且没有确切感染率的报道。根本原因在于,过去没有一种迅速、准确的辅助诊断方法。我们利用基因数据库资料,设计了两对引物,并建立了聚合酶链反应法检测巨细胞病毒技术。该技术反应时间短,结果准确可靠,无交叉反应。我们检测了138例临床诊断为婴儿肝炎综合征的患儿,其中75例检出尿中带病毒,阳性率为66.96％。对照组正常婴儿34名,仅5名尿中查到病毒,阳率性为14.70％。具显著差异(P<0.01)。实验证明,聚合酶链反应法辅助诊断巨细胞病毒感染性婴儿肝炎综合征可靠、可行。数据表明,巨细胞病毒感染在婴儿肝炎综合征中占首要地位。     

肾移植及骨髓移植受者术后巨细胞病毒早早期抗原的检测

背景:肾移植及骨髓移植后,随着免疫抑制剂的应用,潜伏的巨细胞病毒都将被激活。因此早期准确诊断巨细胞病毒感染具有重要的意义,及时而有效地控制巨细胞病毒感染的关键是在血液中迅速、定量地检测到巨细胞病毒的复制。目的:检测肾移植及骨髓移植受者术后的巨细胞病毒早早期抗原,探讨其临床诊断价值。方法:采用免疫组织化学方法对1215例肾移植及骨髓移植受者外周血白细胞巨细胞病毒活动性感染的早早期抗原进行检测。结果与结论:巨细胞病毒早早期抗原指数阳性492例(占40.0%),早早期抗原指数阴性723例(占60.0%),巨细胞病毒抗体IgG明显升高318例(占26.0%),巨细胞病毒抗体IgM90例(占7.0%)。巨细胞病毒抗原血症检测技术对于巨细胞病毒活动性感染的诊断具有高度的灵敏性和特异性,同时对巨细胞病毒的发生还具有重要的预测意义,是一种经济可行的诊断方法。     

人巨细胞病毒在广西不同人群中的感染情况及其与肾脏疾病关系的初步研究

目的 调查广西壮族自治区孕妇、患病新生儿、肾病患者、肾移植患者及正常体检者尿中人巨细胞病毒阳性率,了解人类巨细胞病毒在广西不同人群中的分布情况,初步探讨人巨细胞病毒感染与肾脏疾病的关系.方法 用荧光定量PCR检测研究对象晨尿中人巨细胞病毒DNA.结果 孕妇、患病新生儿、肾病者、肾移植患者及正常体检者尿中人巨细胞病毒DNA阳性率依次为8.18%、3.45%、18.54%、25.42%、0.56%,其在各类肾病患者中的感染率均在10%左右,尤其在狼疮肾炎患者中的感染率更是达到27.78%.结论 人巨细胞病毒在黄疸和早产新生儿中的感染率均较高,需要加强母婴人巨细胞病毒感染的防控工作.人巨细胞病毒很可能在肾病患者发病时对患者肾脏造成损害,并且加重肾病患者的病情,成为肾衰的一个诱因.     

人巨细胞病毒在广西不同人群中的感染情况及其与肾脏疾病关系的初步研究

目的 调查广西壮族自治区孕妇、患病新生儿、肾病患者、肾移植患者及正常体检者尿中人巨细胞病毒阳性率,了解人类巨细胞病毒在广西不同人群中的分布情况,初步探讨人巨细胞病毒感染与肾脏疾病的关系.方法 用荧光定量PCR检测研究对象晨尿中人巨细胞病毒DNA.结果 孕妇、患病新生儿、肾病者、肾移植患者及正常体检者尿中人巨细胞病毒DNA阳性率依次为8.18%、3.45%、18.54%、25.42%、0.56%,其在各类肾病患者中的感染率均在10%左右,尤其在狼疮肾炎患者中的感染率更是达到27.78%.结论 人巨细胞病毒在黄疸和早产新生儿中的感染率均较高,需要加强母婴人巨细胞病毒感染的防控工作.人巨细胞病毒很可能在肾病患者发病时对患者肾脏造成损害,并且加重肾病患者的病情,成为肾衰的一个诱因.     

人巨细胞病毒在广西不同人群中的感染情况及其与肾脏疾病关系的初步研究

目的 调查广西壮族自治区孕妇、患病新生儿、肾病患者、肾移植患者及正常体检者尿中人巨细胞病毒阳性率,了解人类巨细胞病毒在广西不同人群中的分布情况,初步探讨人巨细胞病毒感染与肾脏疾病的关系.方法 用荧光定量PCR检测研究对象晨尿中人巨细胞病毒DNA.结果 孕妇、患病新生儿、肾病者、肾移植患者及正常体检者尿中人巨细胞病毒DNA阳性率依次为8.18%、3.45%、18.54%、25.42%、0.56%,其在各类肾病患者中的感染率均在10%左右,尤其在狼疮肾炎患者中的感染率更是达到27.78%.结论 人巨细胞病毒在黄疸和早产新生儿中的感染率均较高,需要加强母婴人巨细胞病毒感染的防控工作.人巨细胞病毒很可能在肾病患者发病时对患者肾脏造成损害,并且加重肾病患者的病情,成为肾衰的一个诱因.     

大庆地区5700例孕前及孕早期女性巨细胞和风疹病毒感染状况调查

目的分析大庆地区孕前及孕早期妇女风疹与巨细胞病毒的感染状况,为孕前门诊、遗传和优生咨询门诊指导优生,降低先天畸形儿出生率提供理论依据。方法对5700例孕前及孕早期女性开展风疹病毒lgG和IgM,巨病毒IgG和IgM抗体检测,分析孕前和孕早期女性巨细胞病毒与风疹病毒的感染率。结果巨细胞病毒不同年龄段间易感率差异无统计学意义(P0.05),而不同年龄段对象风疹的易感率差异有统计学意义(P0.01),年龄在35岁~39岁易感率最高,为13.66%。结论应加强对高危年龄段孕妇的筛查。对于筛查结果IgM为阳性的孕妇应采取羊水、脐血或绒毛组织进行胎儿宫内感染的产前诊断。     

巨细胞病毒、风疹病毒、弓形虫在已婚育龄妇女中的检测结果分析及调查

目的了解已婚育龄妇女巨细胞病毒、风疹、弓形虫感染情况.方法于2001年5月至2003年10月由我站对全县已婚育龄妇女进行查体,同时抽取血液,采用ELISA方法对巨细胞病毒、风疹病毒、弓型虫抗体进行测定.同时对筛查对象作有关情况调查.结果本县已婚妇女巨细胞病毒感染率6.4%、风疹病毒感染率2.7%.弓形虫感染率6.1%.与正常妊娠组比较,已婚未育组感染率有显著性差异(P＜0.05),异常妊娠组有极显著性差异(P＜0.01),且有动物接触史及有生食习惯者感染率明显高于无动物接触史及无生食习惯者(P＜0.05).结论巨细胞病毒、风疹、弓形虫感染是引起不良生育的主要原因之一,建议已婚育龄妇女孕前加强筛查工作,早诊断、早治疗,确保生1个健康聪明的下一代.     

一例致病机制不明甲型血友病家系的植入前检测

目的对一致病机制不明的甲型血友病家系进行胚胎植入前遗传学检测,以降低该家系生育患儿的风险。方法应用测序、F8基因的1号和22内含子倒位检测及MLPA检测等手段检测已生育甲型血友病患儿的女性。应用植入前检测的方法选择染色体正常的女性胚胎植入并进行产前诊断及随访。结果在已生育甲型血友病患儿的女性外周血样本中检测到F8基因意义未明突变,其它检测未见异常。根据胚胎植入前检测结果,为该女性移植一枚染色体正常的女性胚胎,产前诊断及生后随访确认胎儿为非甲型血友病女性。结论本研究未能明确该家系致病位点,但由于甲型血友病为X连锁隐性遗传病,在对患者充分知情告知并同意后,可应用植入前检测的方法选择女性胚胎移植,以最大限度降低后代患病风险。     

活动性巨细胞病毒感染与反复自然流产的关系探讨

目的探讨活动性人巨细胞病毒（human cytomegalovirus,HCMV）感染与反复自然流产（recurrent spontaneous abortion,RSA）的关系.方法采集反复自然流产孕妇和正常产前体检孕妇外周血,分离外周血单个核细胞（PBMCs）和血浆,分别用免疫荧光法和实时定量PCR检测HCMV pp65抗原和HCMV DNA,并比较2种方法的一致性.结果 65例RSA患者HCMV pp65抗原有20例阳性,阳性率30.8%,50例正常体检孕妇 HCMV pp65抗原有4例阳性,阳性率8.0%,2组孕妇HCMV活动性感染率有显著性差异（χ^2=8.87,P＜0.01）.孕妇HCMV pp65抗原阳性率升高,孕妇流产几率增加（χ^2=7.53,P＜0.01）. 免疫荧光法和实时定量PCR有较好的一致性（92.3%）.结论反复自然流产孕妇 HCMV活动性感染率显著高于正常孕妇,HCMV pp65抗原检测也许可作为RSA早期诊断指标之一.     

应用PCR技术对孕妇HCMV感染的前瞻性研究

应用聚合酶链反应（ＰＣＲ）技术对１５６ 例孕妇尿中ＨＣＭＶ 进行检测,并追踪观察１１４ 例新生儿脐血中ＨＣＭＶ 垂直传播情况。结果表明：１１ 例孕妇尿ＨＣＭＶＤＮＡ 检测阳性,孕期感染率为７－０５％ ,有异常妊娠史孕妇感染率（１４－７１ ％ ）较正常孕妇（４－９２ ％） 明显升高。９ 例ＨＣＭＶ 阳性孕妇有４ 例观察新生儿脐血ＨＣＭＶＤＮＡ 阳性,占４４％ ,而ＨＣＭＶＤＮＡ阴性孕妇无１ 例新生儿脐血出现阳性结果。我们认为ＰＣＲ方法是检测ＨＣＭＶ宫内感染的可靠而灵敏的指标,对ＨＣＭＶＤＮＡ阳性的孕妇应进一步检查羊水,如羊水ＨＣＭＶＤＮＡ也阳性,应终止妊娠。     

Predictive value of maternal-IgG avidity for congenital human cytomegalovirus infection.

BACKGROUND: Human cytomegalovirus (HCMV) is now the most common cause of viral intrauterine infection. Fetal damage is mostly linked to maternal primary infection. It is therefore important to differentiate primary from recurrent or persistent HCMV infection in pregnant females. For this purpose, IgM tests are not reliable enough and the measurement of the IgG avidity appears to be presently the best method. OBJECTIVE: To evaluate the performance of the measurement of HCMV-IgG avidity by a 8 M urea denaturation assay in predicting congenital infection in the offspring. STUDY DESIGN: Seventy-eight women were included in this study on the basis of a HCMV IgM positive or equivocal result on a first serum during pregnancy, but without a documented seroconversion history. The IgG avidity was measured and correlated with the outcome of the pregnancy. RESULTS: In eight cases of HCMV in utero infection the maternal HCMV-IgG avidity index was below 50%. One case of HCMV in utero infection was observed despite a high avidity index during the second trimester of the pregnancy. High or intermediate HCMV-IgG avidity indexes during the first trimester of pregnancy were not associated with a congenital infection. CONCLUSIONS: Even in the presence of an IgM positive result, an HCMV IgG avidity index above 65% on a serum obtained during the first trimester of pregnancy could reasonably be considered as a good indicator of past HCMV infection. In these conditions invasive prenatal diagnosis is not necessary.     

孕妇血清中人类巨细胞病毒DNA含量与胎儿宫内感染的关系

目的探讨孕妇血清中人类巨细胞病毒DNA(HCMV DNA)含量与胎儿宫内感染发生率的关系.方法用PCR方法结合荧光探针的体外扩增和检测技术,检测186例HCMV感染孕妇及胎儿或新生儿血清、胚胎、绒毛中HCMVDNA拷贝数.结果胎儿发生宫内感染及妊娠不良结局与其母素血清HCMV DNA含量有关,随着孕妇血清中HCMV DNA含量的增高,胎儿宫内感染及不良妊娠结局的危险性也呈显著增高趋势.结论孕妇血清HCMV DNA含量升高是胎儿感染HCMV并发生妊娠不良结局的重要因素之一.     

人巨细胞病毒感染对腮腺导管上皮细胞角蛋白K8和K18表达的影响

目的 研究人巨细胞病毒(HCMV)感染对腮腺导管上皮细胞角蛋白K8和K18表达的影响.方法 用免疫组化方法研究腮腺巨细胞包涵体病(PCID)石蜡包埋组织中巨细胞病毒立即早期抗原和角蛋白K8、K18的表达.结果 PCID腮腺组织导管上皮出现包涵体巨细胞;包涵体巨细胞HCMV抗原DDG9/CCH2阳性;包涵体巨细胞角蛋白K8表达呈阴性,而K18表达呈强阳性.结论 HCMV感染腮腺导管上皮细胞诱发角蛋白K8降解,K18表达上调是一种反应性改变;单层上皮角蛋白网具有维持上皮细胞机械力学完整性的功能.     

An antigen fragment encompassing the AD2 domains of glycoprotein B from two different strains is sufficient for differentiation of primary vs. recurrent human cytomegalovirus infection by ELISA.

Primary human cytomegalovirus (HCMV) infection during pregnancy is a frequent cause of fatal damage in populations with low prevalence of HCMV. Differentiation of primary vs. recurrent HCMV infection is an important issue in prenatal counseling. Antibodies specific for viral glycoproteins become detectable only with considerable delay with relation to HCMV infection or IgG seroconversion. Thus, lack of glycoprotein specific (gp-specific) antibodies can serve as a convenient indicator to identify those pregnant women that bear an elevated risk for HCMV transplacental transmission and fetal sequelae. In the opposite case, presence of gp-specific antibodies virtually excludes HCMV primary infection several weeks before sampling. However, no standardized screening assay for HCMV gp-specific antibodies had been available thus far. For this reason, an ELISA based on procaryotically expressed fragments of HCMV glycoprotein B (gB; gpUL55) was developed. Small fragments of gB from two different laboratory strains, encompassing the antigenic domain 2 (AD2) sufficed for sensitive and specific detection of gp-specific antibodies. The gB-ELISA titers correlated with titers of virus neutralizing antibodies in serum samples from primary or recurrent HCMV infections. Seroconversion kinetics of the gB-ELISA in samples from patients with primary HCMV infection closely paralleled the delay in seroconversion of gp-specific antibodies as determined by neutralization assay. Thus this assay provides a diagnostic tool that is easy to perform and can significantly add to available methods for the timely identification of primary HCMV infection during pregnancy. In addition, the gB-ELISA may be helpful in other clinical settings for the differentiation of primary HCMV infection from diseases caused by other pathogens.     

温州市区育龄妇女孕前巨细胞病毒感染现状调查

目的了解温州地区育龄妇女孕前人巨细胞病毒（HCMV）感染的状况。方法收集2008年10月至2010年6日参加温州市龙湾区免费孕前优生筛查的妇女血标本2869份,采用酶联免疫吸附试验（ELISA）检测血清HCMV IgG/IgM抗体;HCMV IgM抗体阳性标本,采用实时荧光定量聚合酶链反应（FQ-PCR）检测血HCMV DNA载量;HCMV IgG/IgM抗体双阳性标本,采用尿素变性结合ELISA技术检测IgG抗体亲和力指数（AI）。结果 2869份孕前妇女血清中HC-MV IgG抗体阳性检出率为97.77%（2805/2869）,HCMV IgM抗体阳性检出率为0.77%（22/2 869）,IgG/IgM抗体均阳性检出率占0.17%（5/2 869）;22份HCMV IgM阳性标本中,血HCMV DNA阳性检出率为68.18%（15/22）;5份HCMVIgG/IgM双阳性标本中,检出低亲和力IgG抗体1份,中等亲和力IgG抗体2份,高亲和力IgG抗体2份。结论温州市区育龄妇女孕前HCMV IgG抗体阳性率高;对HCMV IgM抗体阳性孕前妇女应进行多指标检测以判断HCMV感染的状态,为减少出生缺陷、做好优生优育服务提供依据。     

Role of prenatal diagnosis and counseling in the management of 735 pregnancies complicated by primary human cytomegalovirus infection: A 20-year experience

BackgroundThe burden of congenital human cytomegalovirus (HCMV) infection is well recognized. However, screening for maternal infection remains controversial in view of diagnostic challenges, counseling difficulties, and absence of medical treatment.ObjectiveTo assess the role of prenatal diagnosis and counseling in the management of pregnancy complicated by primary HCMV infection.Study designRetrospective study aimed at investigating diagnostic features, options, and pregnancy outcome in 735 women with primary HCMV infection over a period of 20 years (1990–2009).ResultsOverall, 25.6% women were found to be seronegative before the actual pregnancy. However, none were informed about HCMV infection and potential prevention strategies. Diagnosis of primary HCMV infection was achieved by seroconversion in 44.4% cases and by different combinations of virus-specific IgM, low IgG avidity, and DNAemia in 43.9% cases. Non-specific symptoms and/or haematological/biochemical alterations were recalled by 73.5% women. The onset of infection could be established, and counseling adjusted accordingly in >90% cases. The overall rate of vertical transmission was 37.1%, ranging from 5.6% for preconceptional infections to 64.1% for third trimester infections. Amniocentesis was chosen by 43.1% women, whereas pregnancy termination was requested by 15.6%.ConclusionsReference virology centers and ad hoc trained and experienced physicians are required for accurate diagnosis of primary infection in pregnancy and ensuing counseling. Prenatal diagnosis has a central role in the management of pregnancies complicated by primary HCMV infection. HCMV-seronegative women should receive adequate information.     

Regulation of Abortion by γλ T Cells

PROBLEM: T cells are present at the feto-maternal interface, but their function during pregnancy has not been fully elucidated. T cells bearing γλ T-cell receptor (TCR) may be particularly important, as some subsets can react to trophoblast cells by producing cytokines, such as interleukin-2 (IL-2). METHOD: We depleted T cells bearing the γλ receptor by injecting monoclonal antibodies (mAB) into females of the abortion-prone animal model CBA x DBA/2. We investigated the percentage and number of γλ T-cell receptor positive (TCR)⁺ cells in decidua and spleen during pregnancy in control and γλ-depleted female mice. Pregnant females were also exposed to ultrasonic sound stress to boost the abortion rate. RESULTS: Stress failed to increase the abortion rate in the γλ TCR-depleted mice. FACScan analysis show that the ratio of cells bearing the γλ TCR dramatically decreased after injection of mAB to the γλ TCR in spleen and decidua, these cells recovered six days after depletion, showing a change in cytokine pattern. Levels of TNF-α in decidual γλ T cells decreased; similar effects of decreasing Th₁ cytokines could be observed in splenic γλ T cells. We further identified increased levels of intracellular TNF-α in the Vλ4 subset in the decidua, compared to spleen. CONCLUSIONS: Trophoblast recognition by the Vλ4 T-cell subset in the decidua may cause the release of abortogenic cytokines such as TNF-α. Depletion of such γλ TCR T cells during early pregnancy may promote successful pregnancy outcome in normal pregnancy and prevent stress-induced abortions.     

Real-time PCR quantification of human cytomegalovirus DNA in amniotic fluid samples from mothers with primary infection

A real-time PCR assay was developed to quantify human cytomegalovirus (HCMV) DNA in amniotic fluid (AF) samples collected from 30 pregnant women with primary HCMV infection as detected either from HCMV-immunoglobulin G (IgG) seroconversion or by the presence of HCMV-specific IgG and IgM associated with a low IgG avidity. Clinical information available for each case included ultrasonographic examination and fetal or newborn outcome. HCMV infection of fetuses or newborns was confirmed for the 30 studied cases. AF samples were subdivided into three groups. In group A (n = 13), fetuses presented major ultrasound abnormalities, and pregnancy was terminated. In group B (n = 13), fetuses had normal ultrasound findings, the pregnancy went to term, and the newborns were asymptomatic at birth. In group C (n = 4), fetuses had no or minor ultrasonographic signs, and pregnancy was terminated. The HCMV DNA load values in AF samples were significantly higher in group A (median, 2.8 x 10(5) genome equivalents [GE]/ml) than in group B (median, 8 x 10(3) GE/ml) (P = 0.014). Our findings suggest that HCMV load level in AF samples correlates with fetal clinical outcome but might also be dependent on other factors, such as the gestational age at the time of AF sampling and the time elapsed since maternal infection.