孕妇乙肝免疫球蛋白被动免疫阻断HBV母婴垂直传播作用机理的研究

目的 :研究HBV阳性孕妇孕期应用乙肝免疫球蛋白 (HBIG)预防HBV宫内感染的作用机理。方法 :将 78例乙肝表面抗原 (HBsAg)阳性孕妇分为两组 :预防组 30例 ,于孕 2 8、32、36周肌肉注射HBIG 3次 ,每次 2 0 0IU ;对照组 4 8例 ,只随访查体不用药。检测母儿血清乙肝标志物 (HBVM)和细胞因子IFN γ ,IL 12 ,IL 6水平用双抗夹心酶联免疫吸附法 (DAS ELISA) ,测定HBVDNA含量用荧光定量PCR(FQ PCR)技术。结果 :78例HB sAg阳性孕妇分娩的新生儿宫内感染 10例 ,宫内感染率为 12 .82 % .HBIG预防组孕妇的胎儿HBV感染率显著低于对照组 (P <0 .0 5 ) ;预防组新生儿脐血清抗 HBs检出率显著高于对照组 (P <0 .0 0 1) ;预防组孕妇血清中IFN γ ,IL 12水平显著高于对照组 (P <0 .0 5 ) ,IL 6水平、HBVDNA含量则显著低于对照组 (P <0 .0 5 )。结论 :孕妇HBIG被动免疫可有效阻断HBV母婴垂直传播。     

逆转录聚合酶链反应技术用于产前诊断人巨细胞病毒宫内活动性感染的临床价值

目的　探讨逆转录聚合酶链反应 (RT PCR)技术用于产前诊断人巨细胞病毒 (HCMV)宫内活动性感染的临床价值。方法　应用酶联免疫吸附试验 (ELISA)、RT PCR及血浆套式聚合酶链反应 (N PCR)法 ,检测 2 0 9例孕妇外周血HCMVmRNA、HCMVIgM及HCMVDNA ,其中任意 2项阳性为HCMV活动性感染 (99例 ,观察组 )、任意 2项均阴性为非活动性感染 (110例 ,对照组 )。应用RT PCR和N PCR技术 ,检测两组孕妇从宫内采集的孕早期绒毛共 78例、孕中期脐血和羊水共 131例 ,并追踪胎儿生长发育及出生后情况。结果　 (1)观察组绒毛、脐血HCMV感染率分别为 5 3 3% (16 30 )、4 7 8% (33 6 9) ;对照组分别为 8 3% (4 4 8)、9 7% (6 6 2 ) ,两组比较 ,差异有极显著性 (P <0 0 0 1)。观察组绒毛、脐血HCMV活动性感染率分别为 30 0 % (9 30 )和 2 9 0 % (2 0 6 9) ;显著高于对照组的 4 2 %(2 4 8)和 4 8% (3 6 2 ) ,两组比较 ,差异有极显著性 (P <0 0 0 1)。 (2 )HCMVDNA阳性胎儿中 ,HCMVmRNA阳性胎儿的流 (早 )产率为 4 1 2 % (14 34) ,显著高于HCMVmRNA阴性胎儿的 8 0 % (2 2 5 )。HCMV活动性感染胎儿中 ,有 10例因胎儿生长发育异常而行终止妊娠术。结论　应用RT PCR技术产前诊断HCMV宫内活动性感染安全可靠 ,对于活动性感     

孕妇与胎儿巨细胞病毒感染的研究

目的 :探讨孕妇巨细胞病毒 (CMV)感染 ,早期诊断胎儿CMV感染。方法 :用酶联免疫法 (ELISA)和多聚酶链反应 (PCR)技术检测孕妇血中CMV特异性抗体及CMVDNA ,诊断孕妇CMV感染 ;检测羊水或脐血中CMVDNA诊断胎儿CMV感染。结果 :15 64例孕妇血清中CMV IgM阳性 2 9例 (占 1.8% ) ,CMV IgG阳性 130 6例 (83.5 % ) ,CMVDNA阳性 12 6例 (8.1% ) ;CMV IgM阳性者其CMVDNA均阳性。CMVDNA阳性的 12 6例孕妇其羊水或脐血中CMVDNA阳性 5 2例 (占 4 1.2 % ) ,CMV感染胎儿中有 5例胎儿畸形、2例死胎、3例IUGR ,出生时无明显症状的婴儿中 3例生后 1个月内患黄疸性肝炎 ,1例患新生儿肺炎。 1例发现室间隔缺损、2例出现单侧耳聋。结论 :孕妇CMV感染可造成胎儿严重危害 ,孕妇CMV感染后取羊水或脐血检测CMVDNA是诊断胎儿及新生儿CMV感染的最佳方法。     

孕妇乙型肝炎病毒感染状态与胎儿宫内感染的关系

目的 :探讨孕妇乙型肝炎病毒 (HBV)感染状态与胎儿宫内感染发生率的关系。方法 :用酶联免疫吸附试验(ELISA)筛选出 79例HBV感染孕妇 ,并用荧光定量聚合酶链反应 (FQ PCR)技术检测孕妇血清及脐血中HBV DNA。结果 :4 7例大三阳孕妇血清HBV DNA检出率为 97 9% ,32例小三阳孕妇血清HBV DNA检出率为 9 4 % ,两组差异有非常显著性 (P <0 0 1)。大三阳孕妇脐血中HBV DNA检出率为 31 9% ,小三阳孕妇脐血中HBV DNA检出率为 0 % ,两组差异有非常显著性 (P <0 0 1)。结论 :胎儿宫内感染与孕妇HBV感染状态和HBV DNA检出率有关。     

用荧光探针定量PCR技术检测孕妇与胎儿乙肝病毒感染的研究

目的 :探讨孕妇血中乙肝病毒 (HBV)DNA数量与胎儿乙肝病毒感染的关系。方法 :对 71例孕妇乙肝病毒表面抗原 (HBsAg)阳性者 ,用荧光探针定量PCR(FQ -PCR)方法 ,检测血清、宫颈分泌物及乳汁中HBVDNA数量 ,HBVDNA阳性者肌注乙肝免疫球蛋白 ,分娩后新生儿取末梢静脉血进行HBVDNA定量测定。结果 :孕妇血中HBVDNA阳性者其宫内感染率为 32 .6 % ,宫内感染与孕妇血中HBVDNA数量有关 ,HBVDNA数量高易导致胎儿宫内感染 ,乙肝免疫球蛋白可阻断HBV宫内传播。结论 :FQ -PCR方法能快速、准确检测孕妇血中HBVDNA数量 ,孕妇血中HBVDNA数量高易导致胎儿宫内感染 ,孕期肌注乙肝免疫球蛋白可减少胎儿乙肝病毒感染     

胎儿感染乙型肝炎病毒的临床研究

目的　探讨临床诊断胎儿感染乙型肝炎病毒 (HBV)的方法 ,及其与各种临床因素的相互关系。方法　采用聚合酶链反应 (PCR)及斑点杂交法 ,检测 14 1例HBV携带者孕妇及其分娩的 14 4例新生儿静脉血HBVDNA、HBV标志物及肝功能。其中 4 0例新生儿同时留取脐带血及出生后 2 4～4 8h外周静脉血用于检测HBVDNA。 14 4例新生儿根据有无HBV感染分为胎儿感染组及对照组 ,比较两组新生儿的临床资料及其肝转氨酶水平。结果　 (1) 14 4例新生儿中有 33例发生宫内HBV感染(胎儿感染组 ) ,感染率为 2 2 9% ;无宫内HBV感染 111例 (对照组 )。 4 0例新生儿脐血与外周静脉血HBVDNA阳性率相比 ,脐血的假阳性率为 2 0 0 % ,其敏感性、阳性预测值分别为 10 0 0 %、80 0 %。追踪HBV携带者孕妇所分娩的新生儿出生 6～ 9个月后 ,HBVDNA持续阳性者 7例 (7/2 8) ,抗 HBs转阳率为 85 3% ;出生时HBsAg阳性者 5例 ,均于 1个月后转为阴性。 (2 )在HBeAg或HBVDNA阳性孕妇中 ,其胎儿感染率分别为 70 5 %、6 1 1% ,显著高于HBeAg或HBVDNA阴性者的 2 0 % (2 /10 0 )、0 0 %(P <0 0 1)。胎儿感染组与对照组孕妇 ,在年龄、孕周、分娩方式、出生体重、身长、出生 1分钟Apgar评分等比较 ,差异均无显著性 (P >0 10 )。 (3)胎儿感染组天     

单纯性疱疹病毒Ⅱ型母婴垂直传播初步研究

目的 :研究孕妇及胎儿单纯疱疹病毒 型 (HSV- )的感染及其母婴传播。方法 :采用聚合酶链反应法 (PCR)对 5 74例妊娠晚期孕妇的宫颈分泌物测 HSV- 型 DNA;用酶联免疫吸附法 (ELISA)对其中的 5 3 1例孕妇分娩时抽取母血及新生儿脐血测 HSV- 型Ig G、Ig M抗体。结果 :孕妇血 HSV- 型的 Ig G阳性检出率为 3 7.48%、Ig M阳性检出率为 1 0 .92 % ;胎儿脐血 HSV- 型的 Ig G阳性检出率为 2 9.3 8%、Ig M阳性检出率为2 .6 4 % ;孕妇宫颈分泌物 HSV- 型阳性检出率为 0。新生儿畸形 1例 ,为先天性耳聋。结论 :妊娠晚期孕妇存在 HSV- 的活动感染 ,及母婴宫内垂直传播     

弓形虫感染与妊娠结局的关系

目的　探讨弓形虫 (toxoplasma ,Tox)感染治疗与妊娠结局的关系。 方法　采用ELISA结合PCR法检出Tox感染并要求治疗的孕妇 5 9例及育龄妇女 91例 (治疗组 ) ,均给予口服乙酰螺旋霉素治疗 ,与同期检出Tox感染 ,但未行治疗的孕妇 6 0例和育龄妇女 79例 (对照组 )进行比较 ,比较其Tox IgM及DNA转阴率、宫内垂直传播率及异常妊娠结局发生率。 　结果　治疗组孕妇及育龄妇女外周血中Tox转阴率分别为 83 0 5 % (49/ 5 9)和 70 33% (6 4 / 91) ,对照组孕妇Tox自然转阴率为35 % (2 1/ 6 0 ) ,育龄妇女为 37 97% (30 / 79) ;宫内垂直传播率为 8 4 % (5 / 5 9) ,明显低于对照组的 4 0 %(2 0 / 5 0 ) (χ2 =11 4 970 ,P <0 .0 0 1) ,相对危险度为 0 2 97;随访育龄妇女经治疗Tox转阴后怀孕 2 9例 ,足月分娩 17例 ,脐血Tox IgM、DNA均为阴性 ,体检未发现新生儿异常。　 结论　孕前检测和治疗Tox感染能有效控制胎儿受染 ;孕期治疗能减少孕妇感染 ,降低宫内垂直传播率 ,减少胎儿受损 ,改善妊娠结局。     

乙型肝炎病毒宫内感染机理的研究

目的　探讨乙型肝炎病毒 (HBV)宫内感染的可能机理。方法　应用PCR技术检测 5 9例乙型肝炎病毒表面抗原 (HBsAg)阳性孕妇的羊水、阴道分泌物及其新生儿脐血清中HBVDNA(研究组 ) ,10例乙型肝炎病毒标志物 (HBVM )阴性的正常孕妇及其新生儿为对照组。采用免疫组化ABC染色法检测两组孕妇胎盘组织中的HBsAg及乙型肝炎核心抗原 (HBcAg)的阳性率。 结果　 ( 1)研究组孕妇的羊水、阴道分泌物、新生儿脐血清中HBVDNA阳性率分别为 4 7 5 % ( 2 8/ 5 9)、5 2 5 % ( 31/5 9)、4 5 8% ( 2 7/ 5 9) ;对照组孕妇的羊水、阴道分泌物、新生儿脐血清中均未检出HBVDNA。 ( 2 )研究组孕妇胎盘组织中HBsAg及HBcAg的阳性率 ,呈现出由蜕膜细胞 ( 76 3%及 5 9 3% )、滋养层细胞 ( 72 9%及 5 5 9% )、绒毛间质细胞 ( 6 2 7%及 5 0 8% )至绒毛毛细血管内皮细胞 ( 5 2 5 %及 4 4 1% )依次递减的趋势 ;但其中有 4例孕妇胎盘组织中HBsAg及HBcAg的分布与上述特点相反。研究组孕妇有 32例羊膜细胞中检出HBsAg及HBcAg。对照组孕妇胎盘组织中的HBsAg及HBcAg检出率为零。结论　孕妇血中HBV主要是通过感染胎盘导致胎儿感染 ;但也可能存在胎盘以外的感染途径。     

孕期弓形体感染对胎、婴儿的影响及治疗

目的：研究孕期弓形体感染胎、婴儿的结局，母婴传播途径及治疗。方法：应用ＥＬＩＳＡＩｇＭ方法筛查３８７８例孕妇弓形体近期感染情况。追踪８２例弓形体近期感染孕妇分娩的胎、婴儿预后、收集脐血、羊水、胎盘及尸解的器官组织了解母婴传播途径及病理改变。观察应用螺旋霉素系统治疗孕期弓形体感染者的胎、婴儿预后，并与未治疗组对照。结果：本组孕妇弓形体近期感染发生率为４．４１％（１７１／３８７８），其胎、婴儿出现临床症状为３０．４８％（２５／８２）；孕早期感染以畸胎、流产多见，孕晚期感染表现为早产、低体重儿、黄疸及弓形体感染性肺炎等。从胎盘、羊水、脐血及尸解器官组织中可查出弓形体ＤＮＡ。显示弓形体可通过胎盘感染胎儿，严重者造成器官损害。应用螺旋霉素治疗孕期弓形体感染有效。治疗组先天性弓形体感染发生率低于对照组（Ｐ＜０．０１）。结论：孕妇近期弓形体感染可通过胎盘感染胎儿，造成畸胎、流产、早产、低体重儿及急性弓形体感染症状。螺旋霉素治疗孕期弓形体感染有效。     

用聚合酶链式反应技术检测孕妇与胎儿巨细胞病毒感染的研究

应用聚合酶链式反应（ＰＣＲ）技术检测了孕妇、羊水及脐血中巨细胞病毒（ＣＭＶ）ＤＮＡ。结果表明，１８６例中正常孕妇９８例血清中ＣＭＶＤＮＡ阳性２例。阳性率为２％，异常妊娠（死胎、胎儿畸形及产前咨询）孕妇８８例血清中ＣＭＶＤＮＡ阳性１４例，阳性率为１５．９％，两者差异有显著性，（Ｐ＜０．０１）。提示孕妇ＣＭＶ感染与死胎、胎儿畸形及异常妊娠史有关。通过检测羊水和脐血中ＣＭＶＤＮＡ，发现９例胎儿ＣＭＶ感染，其中３例畸形，２例死胎，１例自然流产，３例足月分娩。     

孕期妇女及胎儿弓形虫感染的研究

弓形虫病是一种人畜共患的寄生虫病，孕妇感染弓形虫可致习惯性流产、死胎、胎儿畸形和出生缺陷，严重危害孕妇和小儿健康。本文采用多聚酶链反应的体外基因扩增技术，对７０例曾有流产史的早期孕妇作绒毛ＰＣＲ检查，结果有１例为阳性，阳性检测率为１．４３％；２０例有不良生育史（头小畸形、脑垂体后叶病、脑发育不全、红斑狼疮等）的中期妊振妇女（５个月以内）作羊水ＰＣＲ检查，有２例为阳性，阳性检测率为１０％；５０例妊娠５个月以上的妇女外用血及她们所生的新生儿脐血作ＰＣＲ检查，未发现阳性结果；对２例检查阳性孕妇作Ｂ型超声波检查，发现１例为无脑儿，１例为脑积水。据此，弓形虫感染直接影响着优生优育。     

Prenatal diagnosis of congenital toxoplasmosis

Ninety-three pregnant women with Toxoplasma gondii seroconversion during pregnancy underwent prenatal diagnosis of fetal toxoplasmosis. The following tests were used: (1). amniocentesis for mouse inoculation (93 subjects), (2). amplification of T. gondii DNA by polymerase chain reaction (PCR) (79 subjects), and (3). cordocentesis for the detection of T. gondii-specific IgM antibodies (13 subjects). All patients had serial ultrasonographic scans to detect those fetuses with abnormalities that could be associated with congenital toxoplasmosis. Eighteen pregnancies (19.4%) had evidence of vertical transmission. A total of 11/18 (61.1%) had positive amniotic mouse inoculation test, while 10/12 (83.3%) had positive PCR results. The combination of both tests allowed the prenatal diagnosis in 17/18 infected fetuses (94.4%). All patients who underwent cordocentesis for the detection of T. gondii-specific IgM antibodies had negative results. However, in two of the above cases fetal toxoplasmosis was detected by amniotic fluid studies. In five of the infected fetuses there were abnormal ultrasonographic findings. All pregnancies with evidence of vertical transmission were terminated, whereas the remaining pregnancies proceeded normally to term. The present data showed that amniotic fluid studies, preferably PCR amplification of T. gondii DNA, are the best diagnostic tools for the detection of vertical transmission in pregnancies with seroconversion during pregnancy.     

检测人巨细胞病毒晚期mRNA在诊断子宫内活动性感染的价值

目的 探讨人巨细胞病毒（ＨＣＭＶ）晚期ｍＲＮＡ检测在宫内活动性感染中的应用价值。方法 采用逆转录－聚合酶链反应（ＲＴＰＣＲ）法对４２例ＨＣＭＶ－ＩｇＭ阳性孕妇外周血及部分胎儿附属物进行ＨＣＭＶ晚期ｍＲＮＡ检测。结果 ４２例ＨＣＭＶ－ＩｇＭ阳性孕妇中,检出晚期ｍＲＮＡ２３便,两者符合率为５４．７％。对其中１３例晚期ｍＲＮＡ阳性孕妇胎儿附属物进行检测,７例阳性,母婴传播率为５３．８％,而１２例晚期ｍＲ     

Incidence of toxoplasmosis in 5532 pregnant women in Crete, Greece: management of 185 cases at risk

OBJECTIVES: To study the incidence of toxoplasmosis in pregnant women in Crete and to test a designed protocol for handling those at risk of delivering congenitally infected infants. STUDY DESIGN: Pregnant women were screened serologically over a period of 5 years. Cases with suspected acute toxoplasmosis were treated, peripheral blood (PB), and amniotic fluid (AF) tested by polymerase chain reaction (PCR) and culture, and fetuses monitored by ultrasonography. The absence of congenital infection in infants was confirmed by serology and clinical evaluation. RESULTS: Of the 5532 pregnant women followed, 70.57% remained seronegative, 29.45% were seropositive, and there was direct evidence of seroconversion in six cases. Acute toxoplasmosis was suspected in 185 cases, maternal parasitemia was detected in five cases and positive amniotic fluid in one case. Congenital infection was excluded in all infants followed, based on the absence of ultrasound findings in utero, lack of clinical symptoms at birth, negative Western blotting (WB) at birth and 3 months later, and descending serology for a year. CONCLUSION: Overall, 29.45% of the pregnant women followed were seropositive, 3.3% with suspected acute toxoplasmosis, and in 0.02% cases there was evidence of maternofetal transmission. The protocol tested allowed differentiation between acute and latent toxoplasmosis, safe management of the cases at risk and assisted in avoidance of unwarranted pregnancy terminations.     

Diagnosis of fetal parvovirus B19 infection: value of virological assays in fetal specimens

Objective  The purpose of our work was to examine the most reliable laboratory diagnosis of fetal parvovirus B19 infection in hydropic fetuses by evaluating the most appropriate clinical sample and laboratory test.
Design  B19 DNA detection in fetal samples and serological signs of B19 infection in the respective mothers. Samples collected between January 2000 and July 2008.
Setting  Microbiology, University of Bologna, Bologna, Italy.
Samples One hundred thirty-five fetal samples (58 fetal cord blood and 77 amniotic fluid samples) and 109 serum samples collected from 109 pregnant women.
Methods  Validated and certified in situ hybridisation assay (ISH) and polymerase chain reaction–enzyme-linked immunosorbent assay (PCR-ELISA) were performed on fetal samples to detect B19 DNA. B19-specific antibodies were investigated in maternal serum samples by a commercial enzyme immunoassay.
Main outcome measures  Parvovirus B19 DNA detection in fetal specimens was analysed in relation to maternal serological signs of infection.
Results  Parvovirus B19 DNA was detected in 22.41% of fetal cord blood and 36.36% of amniotic fluid samples. A statistically significant difference was found between DNA detection by ISH (23.70%) and PCR-ELISA (14.81%) ( P = 0.004). Only 11.76% of fetuses with virological diagnosis of B19 infection were from women with serological signs of acute/recent B19 infection.
Conclusions  Diagnosis of fetal parvovirus B19 infection cannot always rely on maternal serological investigations but rather on the virological analysis of fetal samples. Both fetal cord blood and amniotic fluid samples are suitable for diagnosis, but the detection of B19 DNA in the cells of amniotic fluid samples by ISH proved to be the most reliable diagnostic system.     

正常孕妇,新生儿及异常胎儿血清中人微小病毒B19感染的研究

目的：了解人微小病毒Ｂ１９在母婴中的感染状况以及该病毒对胎儿的危害。方法：应用聚合酶链反应技术（ＰＣＲ）检测３５０例正常孕妇及新生儿血清、９例异常胎儿组织血清人微小病毒Ｂ１９ＤＮＡ。结果：正常孕妇血清Ｂ１９ＤＮＡ阳性率为１．１４％，新生儿脐血阳性率为０．２８％；９例异常胎儿中有６例孕妇血清及脐血Ｂ１９ＤＮＡ阳性（６／９）。应用原位杂交的方法在ＰＣＲ阳性的异常胎儿脑和脾组织中也检出了Ｂ１９ＤＮＡ阳性颗粒。结论：应用ＰＣＲ法检查孕妇及新生儿Ｂ１９病毒具有高度敏感、简便、特异的特点，而原位杂交方法可对病毒感染进行定位，以了解感染部位的病理变化。     

Usefulness of quantitative polymerase chain reaction in amniotic fluid as early prognostic marker of fetal infection with Toxoplasma gondii

OBJECTIVE: Our purpose was to evaluate Toxoplasma gondii concentration in amniotic fluid (AF) samples as a prognostic marker of congenital toxoplasmosis. STUDY DESIGN: A retrospective study was carried out in 88 consecutive AF samples from 86 pregnant women, which were found positive by prospective polymerase chain reaction (PCR) testing. Parasite AF concentrations were estimated by real-time quantitative PCR and analyzed in relation to the clinical outcome of infected fetuses during pregnancy and at birth, taking into account the gestational age at maternal infection. RESULTS: A significant negative linear regression was observed between gestational age at maternal infection and T gondii DNA loads in AF. After adjusting for time at maternal seroconversion by multivariate analysis, higher parasite concentrations were significantly associated with a severe outcome of congenital infection (odds ratio [OR]=15.38/log (parasites/mL AF) [95% CI=2.45-97.7]). CONCLUSION: PCR quantification of T gondii in AF can be highly contributive for early prognosis of congenital toxoplasmosis. Maternal infections acquired before 20 weeks with a parasite load greater than 100/mL of AF have the highest risk of severe fetal outcome.     

CMV Infection and Pregnancy

Cytomegalovirus (CMV) occurs in 0.2?% to 2.2?% of all live births and is the most common cause of intrauterine infection and the leading infectious cause of sensorineural hearing loss and mental retardation. This article reviews literature that relate to the pathogenesis, diagnosis, and treatment of this disease for pregnant women and their fetus. Primary maternal CMV infection during pregnancy has a much higher rate of mother-to-fetus transmission and causes symptoms at birth and long-term disability than nonprimary infection. In addition, some research has shown that children with congenital CMV infection following first-trimester maternal infection are more likely to have severe sequelae. The prenatal diagnosis of fetal CMV infection includes serological testing (IgM detection and IgG avidity assay), amniocentesis, and ultrasound examination. The combination of the presence of CMV IgM antibodies and low CMV IgG avidity, along with maternal or fetal symptoms is used for the diagnosis of a primary maternal infection. Amniocentesis should be complemented until approximately 20-21?weeks of gestation to increase the sensitivity. Because ultrasound abnormalities are only found in less than 25?% of infected fetuses, ultrasound is as a relatively poor predictor of symptomatic congenital infection. CMV hyperimmunoglobulin also may be considered when the pregnant women are confirmed as primary CMV infection with low IgG avidity and amniotic fluid is found to contain CMV or CMV DNA. There is no consensus on the benefit of prenatal administration of ganciclovir into the umbilical vein.