应用聚合酶反应技术检测婴儿肝炎综合征巨细胞病毒和乙型肝炎病…

用聚合酶以应技术（ＰＣＲ）检测６０例婴儿肝炎综合征患儿尿人巨细胞病毒（ＨＣＭＶ０ＤＮＡ在清乙型肝炎病毒（ＨＢＶ）ＤＮＡ，同时做尿ＨＣＭＶ病毒分离。结果表明：ＨＣＭＶ ＤＮＡ阳性４７例，阳性率７８．３％，而ＨＣＭＶ分离阳性３０例，阳性率５０．０％。ＨＢＶ ＤＮＡ阳性１２例，阳性一率２０％。提示婴儿肝炎综合征与ＨＣＭＶ和ＨＢＶ感染有关。ＰＣＲ技术特异，敏感，快速及简便，可协助临床医生对婴儿肝炎综合征做     

60例人类微小病毒B19感染患儿的病原血清学检测及特征

目的了解人类微小病毒Ｂ１９（ｈｕｍａｎｐａｒｖｏｖｉｒｕｓ，Ｂ１９）在儿童中的感染情况。方法采用聚合酶链反应（ＰＣＲ）和酶联免疫吸附（ＥＬＩＳＡ）方法，对１９４例住院治疗（大部分来自血液病房）患儿和１００例健康查体儿童的血清标本进行了检测。抗原为作者采用基因工程方法制备的重组Ｂ１９病毒外壳蛋白ＶＰ１和ＶＰ２。结果在１９４份患儿血清标本中，５５份检测出Ｂ１９病毒ＤＮＡ，３０份Ｂ１９病毒特异性ＩｇＭ抗体检测为阳性，３７份Ｂ１９病毒特异性ＩｇＧ抗体检测结果为阳性，阳性率分别为２８．４％，１５．５％和１９．１％，共有６０例患儿存在Ｂ１９病毒的近期感染。在１００份健康查体儿童血清标本中，３份检出Ｂ１９病毒ＤＮＡ，２份Ｂ１９特异性ＩｇＭ抗体检测结果为阳性，１２份Ｂ１９特异性ＩｇＧ抗体检测结果为阳性，阳性率分别为３０％，２０％和１２０％。结论人类微小病毒Ｂ１９在我国儿童中有较高的感染率，能够导致人类多种疾病，应该引起足够的重视     

输血、血浆与新生儿巨细胞病毒感染的临床研究

１９９２年２～８月应用ＰＣＲ法共检测血源（供体）标本７１份，ＣＭＶ－ＤＮＡ阳性１９份占２８．２％，其中全血１３份，阳性４份占３０．７％；血浆５５份，阳性１４份占２５．５％；白蛋白３份，阳性１份。检测患儿（受体）血标本３０份，阳性１０份。研究对象分为４组，Ⅰ组：受体血ＣＭＶ－ＤＮＡ阴性，供体血阳性，１０例。Ⅱ组：受体血与供体血皆阳性，５例。Ⅲ组：受体血阳性，供体血阴性，５例。Ⅳ组：受体与供体血皆阴性（对照组），１０例。３０例于生后３岁追踪随访，４例死亡，余２６例皆无临床症状和体征。Ⅰ组尿ＣＭＶ－ＤＮＡ阳性率明显高于对照组。建议：新生儿尤其是早产儿、低出生体重儿应尽量避免输血和血浆，必须输注者，以输经ＰＣＲ法筛查的ＣＭＶ－ＤＮＡ阴性血源为宜。     

特发性血小板减少性紫癜与人细小病毒B19感染的关系

目的 研究人细小病毒Ｂ１９（ＨＰＶ Ｂ１９）感染与特发性血小板减少性紫癜（ＩＴＰ）的关系。方法 应用巢式聚合酶链反应技术检测８４例ＩＴＰ患儿存档骨髓涂片标本中ＨＰＶ Ｂ１９－ＤＮＡ,并以２４例巨幼细胞性贫血（ＭＡ）患儿为对照。结果 ８４例ＩＴＰ骨髓中ＨＰＶ Ｂ１９－ＤＮＡ阳性３６例,阳性率为４３％,２４例ＭＡ患儿骨髓中ＨＰＶ Ｂ１９－ＤＮＡ阳性３例,阳性率为１２％,（χ＾２＝６．１９９,Ｐ〈０．０     

套式聚合酶链反应加限制酶分析检测母婴巨细胞病毒感染

为评价套式聚合酶链反应（套式ＰＣＲ）加限制酶分析在孕妇巨细胞病毒感染及其母婴宫内传播检测中的应用，采用套式ＰＣＲ加限制酶分析，病毒分离、电镜观察和特异性抗体测定，对各孕期孕妇外周血，脐血及死胎组织进行人巨细胞病毒（ＨＣＭＶ）检测。结果：３６７名孕妇ＨＣＭＶ阳性检出率为５．５％，其中，套式ＰＣＲ检出率（４．９％）高于病毒分离（３．０％，Ｐ＜０．０５）。６份ＨＣＭＶＤＮＡ阳性母血中，３份配对脐血ＨＣＭＶＤＮＡ也阳性，母－脐传播率为３／６。３对被证实为母－婴宫内传播ＨＣＭＶ的标本中，２对套式ＰＣＲ，病毒分离及特异性ＩｇＭ、ＩｇＡ均阳性，１对套式ＰＣＲ、病毒分离、特异性ＩｇＡ阳性，ＩｇＭ阴性。提示：套式ＰＣＲ能提高诊断ＨＣＭＶ的特异性与敏感性，对孕妇及胎儿／新生儿ＨＣＭＶ感染的研究有重要意义。     

用聚合酶链反应研究先天性畸形与巨细胞病毒感染的关系

应用聚合酶链反应（ＰＣＲ）方法检测畸形和非畸形新生儿死亡病例尸检的石蜡包埋病理组织标本中的巨细胞病毒核糖核酸（ＣＭＶ－ＤＮＡ）片断，研究先天性畸形与宫内ＣＭＶ感染的关系以及胎儿各脏器对ＣＭＶ的易感性。结果显示：（１）畸形组中ＣＭＶ的阳性率高达４２．１９％（２７／６４），明显高于非畸形组的４．６９％（３／６４），差异有非常显著意义（Ｘ￣２＝２５．０７８，Ｐ＜０．００１），提示宫内ＣＭＶ感染与胎儿畸形密切相关；（２）神经系统畸形病例ＣＭＶ的阳性率与循环系统畸形组比较，差异有显著意义（Ｘ￣２＝６．６２６，Ｐ＜０．０５）。提示ＣＭＶ感染可能导致小头畸形、脑积水和脊柱裂等畸形；（３）脑和肝脏是最易受ＣＭＶ感染的脏器；（４）肝胆异常与ＣＭＶ感染有关。本研究说明，先天性ＣＭＶ感染易导致胎儿神经系统崎形及肝脏损害。     

弓形体,人巨细胞病毒和人微小病毒B19感染对新生儿畸形和生长发 …

探讨弓形体（ＴＯＸＯ）人巨细胞病毒（ＨＣＭＶ）及人微小病毒Ｂ１９（ＨＰ－ＶＢ１９）对新生儿畸形及生长发育的影响。采取聚合酶链反应技术对４２例畸形新生儿进行了ＴＯＸＯ－ＤＮＡ，ＨＣＭＶ－ＤＮＡ和ＨＰＶＢ１９－ＤＮＡ的检测，并对检测阳性与阴性畸形新生儿，用标准差记分法（ＳＤＳ）衡量生长发育。ＰＣＲ检测阳性率为１８／４２（４２．８８％），中枢及心血管系统畸形阳性率为１２／１８（６６．６７％），其他系统畸     

新生儿室管膜下囊肿的病因研究

为了探讨新生儿室管膜下囊肿（ＳＥＣ）与先天性感染的关系，采用ＥＬＩＳＡ酶标法检测母婴双方囊肿组与对照组各７０例血巨细胞病毒（ＣＭＶ）、风疹病毒、弓形虫的抗体，并应用聚合酶链反应（ＰＣＲ）技术直接检测病原ＤＮＡ（风疹病毒除外），同时作尿ＣＭＶ的ＰＣＲ检测。结果：囊肿组新生儿血ＣＭＶ－ＩｇＭ抗体和ＣＭＶ－ＰＣＲ阳性率显著高于对照组（分别为１７．１％、５．７％和１２．９％、２．９％）；且尿ＣＭＶ－ＰＣＲ的阳性率达４０．０％（２８／７０），高于对照组的１７．１％（１２／７０）；分别进行两组自身对照，尿ＣＭＶ－ＰＣＲ的阳性率显著高于血ＣＭＶ－ＰＣＲ阳性率（Ｐ＜０．０５）。囊肿组母亲的尿ＣＭＶ－ＰＣＲ阳性率亦显著高于对照组（３０％、１０％、Ｐ＜０．０１）。提示新生儿ＳＥＣ是宫内感染损害中枢神经系统的表现之一，与先天性ＣＭＶ感染有关，尿ＣＭＶ－ＰＣＲ检查可作为宫内ＣＭＶ感染的首选实验室诊断方法。     

新生儿高胆红素血症与人巨细胞病毒感染的关系

为了解高胆红素血症（简称高胆）新生儿中人巨细胞病毒（ＨＣＭＶ）感染状况。方法：用聚合酶链反应（ＰＣＲ）技术对１２０例１２～２８天高胆新生儿的血、尿进行ＨＣＭＶ－ＤＮＡ序列测定。结果：因出生有窒息史、感染致高胆的新生儿中ＨＣＭＶ－ＤＮＡ阳性率分别为３．１％（１／３２）,８．１％（３／３７）。因高胆入院而找不到任何原因的数生儿占１２０例高胆新生儿的１９．２％,与高胆低出生体重儿的ＨＣＭＹ－ＤＮＡ阳性率     

巨细胞病毒肝炎患儿细胞免疫功能变化和干扰素的临床疗效

目的研究巨细胞病毒（ＨＣＭＶ）肝炎患儿细胞免疫功能是否受损；经治疗后临床症状恢复期中免疫功能的恢复情况；干扰素对巨细胞病毒肝炎的疗效。方法用ＤＮＡ探针斑点原位杂交，酶兔方法检查ＨＣＭＶ抗原，尿中分离ＨＣＭＶ３种方法诊断ＨＣＭＶ肝炎。用ＩＬ－２活性测定、Ｔ细胞亚群测定检测患儿细胞免疫功能，将患儿分为干扰素加维生素Ｃ、肌苷治疗组（γＩＦＮ）和仅用维生素Ｃ、肌苷治疗组（非γＩＦＮ），连续观察３个月临床症状恢复情况。结果ＨＣＭＶ肝炎患儿有明显的细胞免疫功能损伤，且临床症状缓解后部份免疫功能尚不能完全恢复。干扰素治疗组的临床症状恢复和第１月与第２月的痊愈率明显优于无干扰素治疗组。黄疸消退平均提前１４．９天（Ｐ＜０．０５）；肝肿大恢复正常提前２８．４天（Ｐ＜０．０１），ＳＧＰＴ恢复正常平均提前１４．９天（Ｐ＜０．０１）。两组的第１月与第２月痊愈率分别为４８．６％与２０．６％、８８．６％与４７．１％（Ｐ＜０．０５）。结论ＨＣＭＶ患儿细胞免疫功能均受损，且不能随症状好转而立即全部好转。干扰素能使ＨＣＭＶ患儿的临床症状加快消退。     

Congenital cytomegalovirus infection in preterm and full-term newborn infants from a population with a high seroprevalence rate

BACKGROUND: Cytomegalovirus (CMV) is the most frequent cause of congenital infections in humans. Prematurity occurs in as many as 34% of infants with symptomatic congenital CMV infection. OBJECTIVE: To determine the clinical presentation and frequency of congenital CMV infection among preterm infants and full-term infants from a population with a high seroprevalence rate. DESIGN/METHODS: A total of 289 preterm infants (median gestational age, 34 weeks; median birth weight, 1,757 g) and 163 term infants (median gestational age, 39 weeks; median birth weight, 3,150 g) sequentially born were included in the study. Serum IgG antibodies to CMV were measured in all mothers. One urine sample was collected within the first 7 days of age from all newborns. Virus isolation in urine samples was performed by tissue culture, and viral DNA was detected by a multiplex PCR. CMV infection was diagnosed in infants with virus excretion detected by both methods on at least two occasions within the first 3 weeks of life. RESULTS: Maternal CMV seropositivity rate was 95.7%. Congenital CMV infection was detected in 6 of 289 (2.1%) (95% confidence interval, 0.84 to 4.68) preterm infants and in 3 of 163 term infants (1.8%) (95% confidence interval, 0.48 to 5.74) (P > 0.05). Four of 6 preterm infants with congenital CMV infection were symptomatic, but none of the term infants was symptomatic (P = 0.16). CONCLUSION: The frequency of congenital CMV infection in preterm newborn infants from mothers with a high seropositive rate was similar to that found in term infants. No significant difference was found between the proportion of symptomatic infants among preterm and term infants. Our finding of symptomatic congenital CMV infection underscores the need of further evaluation of correlates of congenital symptomatic infection in highly immune populations.     

先天性及非先天性巨细胞病毒感染的临床研究

目的研究先天性、非先天性巨细胞病毒感染对患儿的损伤及其治疗措施。方法根据国内采用的巨细胞病毒(CMV)感染诊断方案,血CMV-IgM、CMV-pp65检测之一阳性者确定为CMV感染,46例患儿均确诊为CMV感染。按症状开始时间,分为先天性感染组(先天组)和非先天性感染组(包括围生期感染和生后感染,非先天组)。比较两组临床特征,对婴儿肝炎综合征患儿予更昔洛韦治疗,比较治疗前后肝酶及胆红素变化,观察更昔洛韦不良反应,对两组进行随诊,随诊中比较其转归情况。结果先天组21例,非先天组25例。CMV-IgM阳性38/46例,CMV-pp65阳性19/43例,两者均阳性11例。先天组并先天性畸形者较多,两组相比有显著性差异(P<0.05)。对婴儿肝炎综合征患儿予更昔洛韦治疗,治疗后胆红素、肝酶下降明显;更昔洛韦不良反应为粒细胞减少。随诊中发现先天组3例患儿死亡,1例于10个月时出现听觉脑干诱发电位异常。先天性感染可并听力损伤,且可在生后进展。先天性感染及生后感染均可引起肝胆受损,但部分先天性感染患儿预后不良。结论先天性感染较非先天性感染对患儿危害更严重。     

HCMV HSVⅡ和HPVB-19感染对胎儿畸形的影响

目的：研究人巨细胞病毒(HCMV),单纯疱疹病毒Ⅱ型(HSVⅡ)及人微小病毒B19(HPV B19)感染与胎儿畸形的关系。方法：应用聚合酶链反应技术检测了畸形组16例和对照组25例围生期死亡病例的脑、肝、肺、肾、脾组织和妊母胎盘组织中的HCMV,HSVⅡ及HPV B19 DNA。结果：①畸形组HCMV,HSVⅡ,HPV B19阳性率分别为：50.0%(8/16),37.5%(6/16)和31.3%(5/16),而对照组阳性率分别为8.0%(2/25),8.0%(2/25),12.0%(3/25),两组之间CMV,HSVⅡ感染有显著性差异(P<0.05)。②HCMV易感器官为脑、肝、肺,畸形组中脑组织阳性率43.8%(7/16)与肾组织6.3%(1/16)有显著性差异,HSVⅡ易感器官主要为肝脏,HPV-B19主要为脾脏受累。结论：HCMV,HSVⅡ胎儿与畸形有密切关系,HPV-B19感染因例数较少与畸形无明显关系。     

Variable outcome of a congenital cytomegalovirus infection in a quadruplet after primary infection of the mother during pregnancy

A congenital cytomegalovirus (CMV) infection in a quadruplet has not been reported previously in the literature. We describe a congenital CMV infection in a quadruplet after primary CMV infection of the mother during pregnancy. One infant died antenatally, while another had cholestatic jaundice at birth and died of liver failure at three months of age. Of the two surviving infants, one showed no signs or symptoms of congenital CMV infection at 18 months of age, whereas the other had hearing loss and delayed development. CMV cultures of urine were positive at six weeks of age in all three infants born alive. The diagnosis of congenital CMV infection after primary CMV infection of the mother during pregnancy was made retrospectively by detection of CMV-immediate early antigen in three placentas as well as by examination of serum obtained from the mother during pregnancy.     

Urine polymerase chain reaction as a screening tool for the detection of congenital cytomegalovirus infection

OBJECTIVES: To define the incidence of congenital cytomegalovirus (CMV) infection in a defined population in Israel as diagnosed by urine polymerase chain reaction (PCR), and to assess the utility of this method for screening for congenital CMV infection. DESIGN: A convenient sample of urine specimens from asymptomatic newborns were subjected to CMV PCR. Positive results were validated by urine tube culture and by determination of serum CMV IgM antibodies. Maternal CMV IgG was determined in a representative sample of mothers. Newborns with positive urine specimens underwent full clinical evaluation. Epidemiological characteristics of the mothers were extracted from the medical records. SETTINGS: Two medical centres in Israel with different population characteristics. PATIENTS: A total of 2000 newborns (1000 in each medical centre). MAIN OUTCOME MEASURE: Presence of CMV DNA in the urine. RESULTS: Despite significant epidemiological differences between the populations in the two hospitals, the CMV seroprevalence was similar, 80.5% and 85%. Fourteen of the 2000 newborns screened (0.7%) were PCR positive. Urine culture was positive in nine of 10 specimens; IgM was positive in only two of 13 newborns with positive PCR. Eleven newborns underwent full or partial evaluation, and only one (9%) was symptomatic. CONCLUSIONS: The incidence of congenital CMV infection in the study population was 0.7%; over 90% were asymptomatic. Urinary CMV PCR is a reliable, rapid, and convenient method, and thus may serve as a screening tool for the detection of congenital CMV infection.     

定量检测婴儿尿液巨细胞病毒DNA的临床意义

目的 探讨荧光定量聚合酶链反应(FQ-PCR)检测尿液巨细胞病毒(CMV)在儿科CMV感染诊断治疗中的意义.方法 应用FQ-PCR方法检测婴儿尿液CMV DNA量.结果 (1)280例疑似CMV感染儿有126例尿液中CMVDNA呈阳性,阳性率是45.0%,为症状性CMV感染儿;同年龄组的70例健康儿童尿液中CMV DNA阳性5例,阳性率是7.1%,为无症状性CMV感染儿.症状性感染儿尿液中CMVDNA拷贝数对数值(4.78±0.85)与无症状性感染儿(4.45±1.02)之间差异无显著性(P＞0.05).(2)90例症状性CMV感染儿应用更昔洛韦诱导治疗结束时尿液CMV DNA拷贝数对数值(3.74±0.58)较治疗前(4.64±0.90)明显减少(P＜0.01),临床症状改善.(3)45例症状性CMV感染儿在治疗后3个月CMVDNA拷贝数对数值(4.41±0.74)较治疗结束时(3.67±0.58)升高(P＜0.01).临床症状明显改善.结论 FQPCR方法检测婴儿尿液中CMVDNA量可以诊断CMV感染,但是不能区分CMV感染是症状性感染还是无症状性感染;应避免盲目反复应用更昔洛韦治疗CMV感染.     

Congenital cytomegalovirus infection: association between virus burden in infancy and hearing loss

OBJECTIVE: To determine the relationship between the virus burden in infancy and hearing loss in congenital CMV infection. STUDY DESIGN: A cohort of 76 infants with congenital cytomegalovirus (CMV) infection identified by means of newborn virologic screening was monitored for outcome. The amount of infectious CMV was analyzed in urine specimens obtained during early infancy. Peripheral blood (PB) samples obtained during early infancy were available from 75 children and CMV DNA was quantitated with a real-time quantitative polymerase chain reaction. RESULTS: Infants with clinical abnormalities at birth (symptomatic congenital CMV infection) had higher amounts of CMV in urine (P = .005) and CMV DNA in PB (P = .001) than infants with no symptoms. Eight children with and 4 children without symptoms had hearing loss. Among children without symptoms, those with hearing loss had a significantly greater amount of CMV in urine (P = .03) and PB virus burden (P = .02) during infancy than those with normal hearing. Infants with < 5 x 10(3) pfu/mL of urine CMV and infants with < 1 x 10(4) copies/mL of viral DNA in PB were at a lower risk for hearing loss. CONCLUSION: In children with asymptomatic congenital CMV infection, hearing loss was associated with increased amounts of urine CMV and PB CMV DNA during early infancy.     

Hirschsprung's Disease: A Viral Etiology?

This study investigates the hypothesis that Hirschspiung's disease (HD), congenital rectal agangliosis, may be etiologically linked to antenatal cytomegalovirus (CMV) infection. Bowel specimens from 72 HD patients, 144 control I infants (Hirschsprung-like symptoms, normal histology), and 36 control 11 infants (deaths from nongastrointestinal causes) were analyzed for CMV genomes by polymerase chain reaction. Positive results were obtained in 6 HD patients (8.8%) and none of the controls. Our findings suggest that antenatal CMV infection, a potentially preventable condition, may be one of the etiological factors in HD.     

北京地区新生儿先天巨细胞病毒感染状况研究

目的评估北京地区新生儿先天性巨细胞病毒(CMV)感染状态及其对新生儿的危害。方法选择2004年11月至2008年3月在北京居住、怀孕12周之内的孕妇,分别于孕早期和孕中期进行血清CMV IgG定量和CMV IgM定性检测;活动感染者接受CMV IgG亲和力、CMV pp65抗原血症和白细胞CMV巢式聚合酶链反应(nPCR)检测;白细胞CMV DNA阳性孕妇接受羊水CMV nPCR检测。所有入选孕妇分娩的新生儿出生后均检测脐血CMV nPCR,阳性者于生后2周内复查尿CMV nPCR。结果本研究共收入孕妇1752例,新生儿1756例;孕期血清学检查CMV活动感染51例,占2.91%(95%CI 2.12%～3.70%),其中原发活动感染2例,占全部孕妇的0.11%(95%CI 0.10%～0.41%);CMV nPCR阳性的孕期活动性感染占0.34%;活动感染孕妇羊水CMV nPCR检测全部阴性;新生儿先天性CMV感染发生率0.23%(95%CI 0.06%～0.58%)。所有先天性CMV感染新生儿出生时均为无症状性感染。结论北京地区CMV感染垂直传播率低,先天CMV感染发生率0.23%,未见到症状性先天性CMV感染患儿。     

Prevalence of rubella virus and cytomegalovirus infections in suspected cases of congenital infections

Although rubella virus and cytomegalovirus (CMV) are important causes of congenital infections, information on their prevalence in our country is scarce. We studied a total of 249 infants suspected of having congenital infections from January 1988 to September 1989. Serum samples of these infants were tested for rubella and cytomegalovirus specific IgM antibodies by mucapture ELISA. Thirty (12%) infants were positive for rubella IgM antibody, and 50 (20%) had CMV specific IgM antibody. In the group presenting with hepatosplenomegaly (n=56) rubella and CMV specific IgM antibodies were detected in 1 (1.7%) and 25 (44.6%) infants respectively. In the group presenting with congenital malformations (n=90), 23 (25.5%) were positive for rubella, and only 9 (10%) had CMV IgM antibodies. Of the infants presenting with mental retardation (n=39), only CMV infection was detected in 3 (7.7%) infants, whereas amongst the group showing intrauterine growth retardation (n=16), 5 (31.25%) had CMV specific IgM antibodies and 2 (12.5%) had rubella specific IgM antibodies. In the miscellaneous group (n=48), 4 (8.3%) and 8 (16.6%) infants had rubella and CMV IgM antibodies respectively. CMV infection was prevalent in a significantly higher number of children with hepatosplenomegaly than rubella while in infants with congential malformations a significantly higher number had rubella infection. It is concluded that rubella and CMV infections are commonly seen in children with intrauterine infections in our population.