用间接免疫荧光法和PCR技术重新评价抗CMV－抗体检测的临床意义

为了正确评价临床常用的检测抗巨细胞病毒（ＣＭＶ）抗体的临床意义，用间接免疫荧光法（ＩＦＡ）和聚合酶链反应技术（ＰＣＲ）检测了５０例急性黄疸型甲型肝炎患儿血白细胞中ＣＭＶ早期抗原（ＥＡ）和晚期抗原（ＬＡ）、血白细胞及尿沉渣ＣＭＶ－ＤＮＡ，阳性率分别为５８％、２２％、６８％、６２％。同期检测抗ＣＭＶ－ＩｇＧ、抗ＣＭＶ－ＩｇＭ，并观察ＩｇＧ急性期／恢复期是否升高４倍以上（ＩｇＧ↑↑），阳性率分别为８６％、３４％及４６％。仅检测１次ＩｇＧ，１４％的患者会漏诊；动态监测ＩｇＧ，４３．９％的活动性ＣＭＶ感染者会漏判；仅检测１次ＩｇＭ，５８．５％的活动性ＣＭＶ感染者会漏判。７９％的ＩｇＧ阳性患者为活动性感染（ＥＡ、ＬＡ、ＩｇＧ↑↑或ＩｇＭ任何１项阳性）；原发性及继发性活动性ＣＭＶ感染者的ＩｇＧ↑↑、ＥＡ、ＩｇＭ阳性率分别为１００％、４２．９％、２８．６％和４７．０％、７６．５％、４４．１％，表明原发感染诊断主要依靠ＩｇＧ↑↑，而继发活动感染则需多种指标。同期检测了２２名健康ＣＭＶ感染儿童的上述指标，与之无显著差异。     

特异性转移因子对活动性巨细胞病毒感染孕妇的治疗作用

①目的探讨人巨细胞病毒（ＨＣＭＶ）特异性转移因子（ＳＴＦ）对活动性巨细胞病毒感染孕妇的治疗作用。②方法以孕妇血清ＨＣＭＶ特异性ＩｇＭ阳性做为ＨＣＭＶ活动性感染的指标，应用普通转移因子（ＴＦ）、高效价免疫球蛋白（ＨＩＧ）及ＳＴＦ分别对５７例、１８例及４６例感染孕妇进行治疗，以血清ＨＣＭＶ－ＩｇＭ转阴作为有效的指标。③结果ＴＦ组的ＨＣＭＶ－ＩｇＭ转阴率为２８．１％（１６／５７）；ＨＩＧ组的转阴率为５５．５％（１０／１８）；ＳＴＦ组的转阴率为５６．５％（２６／４６）。ＳＴＦ的治疗作用优于ＴＦ（χ２＝９．７８０，Ｐ＜０．０１）；而ＳＴＦ与ＨＩＧ对感染孕妇的治疗作用无显著性差异（χ２＝０．０１，Ｐ＞０．０５）。④结论ＳＴＦ对活动性ＨＣＭＶ感染孕妇具有较好的治疗作用。     

特异性转移因子对活动性巨细胞病毒感染孕妇的治疗作用

目的 探讨人巨细胞病毒（ＨＣＭＶ０特异性转移因子（ＳＴＦ）对活动性巨细胞病毒感染孕妇的治疗作用。方法 以孕妇血清ＨＣＭＶ特异性ＩｇＭ阳性做为ＨＣＭＶ活动性感染的指标，普通转移因子（ＴＦ）、高效价免疫蛋白ＨＩＧ及ＳＴＦ分别对５７例、１８例及４６例感染孕妇进行治疗，以血清ＨＣＭＶ－ＩｇＭ转阴作为有效的指标。结果 ＴＦ组的ＨＣＭＶ－ＩｇＭ转阴率为２８．１％；ＨＩＧ组的转阴率为５５．５％；ＳＴＦ组的转阴率     

孕妇及新生儿感染人巨细胞病毒的检测和临床意义

孕妇感染人巨细胞病毒（ＨＣＭＶ）后，可造成流产、早产、死胎、胎儿宫内发育迟缓、婴儿先天畸形及新生儿病毒性肝炎等。采用ＥＬＩＳＡ法对５５例孕妇及其新生儿进行巨细胞病毒特异性ＩｇＭ和ＩｇＧ抗体检测。结果表明孕妇血清ＩｇＭ和ＩｇＧ抗体阳性率分别为２７．３％和９４．５％；新生儿脐血ＩｇＭ和ＩｇＧ阳性率分别为１０．９％和４５．５％；异常产儿６例中其母亲ＨＣＭＶ特异性ＩｇＭ抗体均为阳性，表明孕妇有ＨＣＭＶ近期感染对新生儿有一定影响。     

初产妇人工流产史与孕期人巨细胞病毒活动性感染的关系

目的探索既往有人工流产史的初产妇与孕期人巨细胞病毒（ＨＣＭＶ）活动性感染的关系。方法用ＥＬＩＳＡ法检测无自然流产及死胎史的初产妇ＨｃＭＶ特异性抗体（ＩｇＭ，ＩｇＡ），共检测１３１７例。其中有人工流产史的３７４例（人流组）和无人工流产史的９４３例〔无人流组）ＨＣＭＶ－ＩｇＭ，ＩｇＡ阳性结果行配对比较；人流组人流的次数与孕期ＨＣＭＶ－ＩｇＭ，ＩｇＡ阳性率的关系比较。结果人流组ＨＣＭＶ活动性感染率２７．５４％，无人流组１８．２４％，差异有显著性（Ｐ＜０．０１）。人流１次，其活动性感染率２４．３５％，２次或２次以上的为３５．８２％，差异有显著性（Ｐ＜０．０５）。结论初产妇孕期ＨＣＭＶ活动性感染与既往人流及反复人流史关系密切     

母乳中人巨细胞病毒的检测及哺乳期妇女感染状况的研究

用人巨细胞病毒（ＨＣＭＶ）单克隆抗体，对２７９例哺乳期妇女的乳汁进行了检测。结果表明：长春市城区母乳中ＨＣＭＶ－ＩｇＧ抗体的阳性率为７７．１％，ＨＣＭＶ－ＩｇＭ抗体的阳性率为１４％，其中包括１３例ＨＣＭＶ－ＩｇＧ、ＨＣＭＶ－ＩｇＭ抗体均阳性，阳性率为４．７％。城区哺乳期妇女ＨＣＭＶ潜伏感染率为７４．９％，原发感染率为１１．５％，再发感染率为４．７％，后两者比较差异非常显著（χ２＝１３９．８，Ｐ＜０．００１）。提示，长春城区母乳ＨＣＭＶ－ＩｇＭ阳性率比我国部分地区低，哺乳期的感染以原发感染为主。     

抗HCV-IgM抗体测定及临床初步应用

将市售ＨＣＶ－ＩｇＧ诊断试剂盒加以改良，建立了ＨＣＶ－ＩｇＭ抗体检测方法，与抗ＨＢｃ－ＩｇＭ，抗ＨＢｓ－ＩｇＭ，抗ＣＭＶ－ＩｇＭ，抗ＲＶ－ＩｇＭ，ＲＦ及抗核抗体（ＡＮＡ）阳性血清均不起反应。正常人群中抗ＨＣＶ－ＩｇＭ阳性率为１．５％（１／６８），抗ＨＣＶ－ＩｇＧ阳性的４３例血透患者检出抗ＨＣＶ－ＩｇＭ１４例（３２．６％），１２例输血后肝炎抗ＨＣＶ－ＩｇＭ全部阳性。以上结果表明，抗ＨＣＶ－ＩｇＭ可作为输血后ＨＣＶ感染的一个敏感指标     

51例病毒性心肌炎患儿血清柯萨奇 B 组病毒特异性 IgM 检测分析

用间接ＥＬＩＳＡ法检测５１例病毒性心肌炎患儿血清中柯萨奇Ｂ组病毒（ＣＶＢ）ＩｇＧ、ＩｇＭ，结果ＣＶＢ－ＩｇＭ≥１∶１００（＋）２１例，占４１．２％。３５例非心肌炎组患儿ＣＶＢ－ＩｇＭ≥１∶１００（＋）２例，占５．７％。经统计学处理，两者有非常显著差异（Ｐ＜０．０１）。说明ＣＶＢ是合肥地区病毒性心肌炎的主要病原；ＣＶＢ－ＩｇＭ检测，方法简便，准确可靠，具有早期诊断价值。ＩｇＭ阳性组病程均在两个月以内，９０．５％有明确的上呼吸道或肠道感染史，ＳＴ－Ｔ改变占２８．６％，与ＩｇＭ阴性组比较差异显著（Ｐ＜０．０５）；ＩｇＭ阳性组心脏扩大、心功能不全均较阴性组多，但经χ２检验没有差异（Ｐ＞０．０５）；ＩｇＭ阳性组与阴性组在心律紊乱、酶学异常上没有差异。     

不育妇女巨细胞病毒感染的研究

目的：探讨妇女不育与巨细胞病毒（ＨＣＭＶ）感染的关系。方法：应用酶联免疫吸附试验（ＥＬＩＳＡ）方法,对９８例育龄妇女的血清进行了巨细胞病毒抗体ＩｇＭ、ＩｇＧ的检测,其中４８例为不育妇女,另５０例为正常对照组。结果：不育组ＨＣＭＶ－ＩｇＧ的阳性率为６２．５％,对照组为７０．０％,两者差异不明显（Ｐ〉０．０５）,但不育组ＨＣＭＶＩｇＭ阳性率为４８．５％,明显高于对照组的４．０％（Ｐ〈０．０５）。对不育     

疱疹病毒经胎盘传播的研究

应用ＰＣＲ技术和ＥＬＩＳＡ间接法对贵阳地区２８６对母婴配对血清进行巨细胞病毒（ＣＭＶ）和单纯疹病毒（ＨＳＶ）ＤＮＡ和特异性ＩｇＭ、ＩｇＭ抗体检测。结果表明贵阳地区产妇ＣＭＶ、ＨＳＶ特异性ＩｇＧ抗体阳性率分别为８５．６７％和８０．７７％;新生儿血清ＣＭＶ、ＨＳＶ特异性ＩｇＧ抗体阳性率为８２．８７％和７６．９２％。而产妇血清ＣＭＶ、ＨＳＶ１,２ＤＮＡ检出率分别为１２．２４％、９．７９％和６．２９％;新     

巨细胞病毒感染与儿童免疫性血小板减少性紫癜的关系探讨

目的探讨人巨细胞病毒(HCMV)感染与儿童免疫性血小板减少性紫癜(ITP)的关系。方法采集154例ITP患儿(ITP组)和50例健康儿童(对照组)的血清样本,用Real-time PCR检测HCMV DNA,ELISA方法检测HCMV IgM、IgG抗体;其中105例ITP患儿和50例健康对照儿童采集了尿液标本,用Real-time PCR检测HCMV DNA,并比较ITP组HCMV DNA阳性患儿与阴性患儿的血小板数量的差异。结果 ITP患儿血清HCMV DNA、HCMV IgM及IgG抗体和尿HCMV DNA阳性率均明显高于健康对照儿童,两组比较差异均有统计学意义(P<0.01);ITP组HCMV DNA阳性患儿的血小板数量(29.72±14.54)×109/L与阴性患儿(41.28±18.35)×109/L比较差异有统计学意义(P<0.05)。结论儿童感染HCMV可能是发生ITP的重要致病因素之一,这对指导临床有效治疗及预防ITP的发生有着积极的意义。     

活动性巨细胞病毒感染患儿免疫状态的变化及其意义

目的研究活动性人巨细胞病毒(HCMV)感染患儿的免疫功能状态。方法以68例活动性HCMV感染患儿为观察对象,44例潜伏型HCMV感染者及40例健康儿童为对照组;采用ELISA检测HCMV-IgM、IgG,用PP65抗原血症监测法测HCMV-Ag,用PCR方法检测尿HCMV-DNA,血清免疫球蛋白测定采用琼脂单扩散法,外周血T细胞亚群检测采用单克隆抗体标记的间接ABC免疫法。结果三组体液免疫功能测定差异无显著性(P>0．05);HCMV 感染组CD3、CD4、CD4/CD8比潜伏型CMV感染组及健康对照组明显降低(P均<0．01)。结论 HCMV感染患儿体液免疫功能基本在正常水平,不起主要作用;HCMV感染主要影响细胞免疫功能,细胞免疫功能低下是HCMV感染存在的结果也是前提条件。因而治疗上给予免疫调节剂是必要的。     

IgG抗体亲和力指数在儿童巨细胞病毒感染诊断中的意义

目的探讨IgG抗体亲和力指数(AI)在儿童巨细胞病毒(HCMV)感染诊断中的意义。方法分别收集120例临床疑似HCMV感染患儿的尿液和外周血标本,采用实时荧光定量聚合酶链反应(FQ-PCR)和微粒子酶免化学发光法检测尿液HCMV DNA载量和血清HCMV IgM/IgG抗体;采用尿素变性结合酶联免疫吸附试验(ELISA)测定HCMV IgG抗体亲和力指数(AI)。结果 120例疑似HCMV感染患儿的尿液HCMV DNA、血清HCMV IgM和HCMV IgG阳性检出率分别为70.83%(85/120)、60.83%(73/120)和76.67%(92/120),差异有统计学意义(χ2=7.252,P=0.027);25例HCMV IgM-/IgG+患儿中,以高亲和力抗体检出为主,占92%(23/25);67例HCMV IgM+/IgG+患儿中,以低、中等亲和力抗体为主,分别占26.87%(18/67)和53.73%(36/67);高、中、低IgG抗体AI患儿尿液HCMV DNA阳性检出率分别为36.11%(13/36)、86.84%(33/38)和100%(18/18),差异有统计学意义(χ2=32.262,P<0.01)。结论 IgG抗体亲和力指数测定能准确地反映HCMV感染的活动状况,有助于HCMV感染的诊断与治疗。     

A clinical study of cytomegalovirus infections during pregnancy

Summary Enzyme-linked Immunosorbent Assay (ELISA) was adopted to detect the human cytomegalovirus (HCMV) specific IgG, IgM in the blood samples of 216 pregnant women and the umbilical cord blood, as well as the HCMV antigen in the urine specimens of 91 newborn infants. Emphasis was put on the occurrence of cytomegalovirus infection during pregnancy and influence on both the mother and the infant. Our experimental results showed that HCMV IgG positive rate in the maternal sera as well as in cord sera was 93.98%; HCMV IgM positive rates were 8.8% in maternal sera and 3.7% in cord sera, respectively. In maternal sera IgG-positive group, 31.6% cord sera showed also HCMV IgG positive results. In 91 urine specimens of newborn infants, HCMV-Ag positive specimens accounted for 10.98% (10 cases). The rates of abortion, congenital malformation, stillbirth, premature fetal death, and intrauterine growth retardation (IUGR) were all apparently higher in the HCMV-IgM positive pregnant group than in the HCMV-IgM negative group. The infants of HCMV-IgM positive mothers also showed lower birth weight, height, biparietal diameter, suboccipitobregmatic diameter, occipitofronatal diameter, occipito-mental diameter than the infants of HCMV-IgM negative mothers. Meanwhile, the growth and development indices of the former showed a tendency to decrease. Placenta histological examination showed thac HCMV-IgM positive group had a higher incidence of pathological changes than HCMV-IgM negative group. Altogether, there were 8 cases of cogenital infections.     

IFN-γ/TL-4在儿童免疫性血小板减少性紫癜中的表达

目的：分析儿童急性特发性血小板减少性紫癜（ITP）血中细胞因子IFN-γ/IL-4的含量,从细胞因子水平探索急性ITP患儿Th1/Th2细胞分化趋势,可望揭示其部分发病机理。方法：应用实验研究策略,选择急性ITP患儿发作期组30例,缓解期组28例,于同群体中选择健康儿童20名为对照组,用ELISA法检测血浆IFN-γ和IL-4含量,进行分析比较。结果：①IFN-γ血浆含量发作期组（269.33±13.03）pg/ml明显高于缓解期组（178.84±7.22）pg/ml和对照组（149.26±10.66）pg/m,且缓解期组亦高于对照组,差异均有统计学意义（P〈0.01,95%可信区间内不包含1）;②IL-4发作期组（12.34±1.14）pg/ml明显低于缓解期组（21.38±1.36）pg/ml和对照组（26.41±1.53）pg/ml,且缓解期组亦低于对照组,差异均有统计学意义（P〈0.01,95%可信区间内不包含1）。结论：血中IFN-γ增高,IL-4降低,显示儿童急性ITP存在明显的Th1优势分化现象。     

乳头瘤病毒及巨细胞病毒在宫颈癌组织中的协同作用

应用ＤＮＡ－ＤＮＡ斑点杂交技术，对６４例宫颈标本（包括正常宫颈２６例，重度宫颈糜烂１７例，宫颈癌２１例）分别检测ＨＰＶ－１６ＤＮＡ及ＨＣＭＶＤＮＡ。结果发现正常宫颈、宫颈糜烂及宫颈癌组织中ＨＰＶ１６ＤＮＡ的检出率分别为２３．０８％、６４．７１％、７１．４３％。而ＨＣＭＶＤＮＡ的检出率则依次为３．８５％、２３．５４％、及４２．８６％，两者检出率呈直线相关（ｒ＝０．９２３）。因此，推测ＨＣＭＶ很可能与ＨＰＶ１６协同作用导致宫颈癌的发生。     

儿童急性特发性血小板减少性紫癜转录因子的表达

目的：分析儿童急性特发性血小板减少性紫癜（ITP）血中Thl／Th2特异性转录因子T-bet／GATA3mRNA的表达,可望揭示其部分发病机理。方法：选择急性ITP患儿发作期组30例,对照组20例。引入RT-PCR技术,测定血中T-bet和GATA3mRNA的表达。结果：①T-betmRNA的表达发作期组明显高于缓解期组和对照组,且缓解期组亦高于对照组,差异均有统计学意义（P〈0．01）。②GATA3mR-NA的表达发作期组明显低于缓解期组和对照组,且缓解期组亦低于对照组,差异均有统计学意义（P〈0．01）。结论：血中T-betmRNA增高,而GATA3mRNA降低,显示儿童急性ITP存在明显的Thl优势分化现象,且可能与转录因子T-bet和GATA3的调控作用有关;T-bet／GATA3与儿童急性ITP的发病有一定的关系。     

DETECTION OF HUMAN CYTOMEGALOVIRUS INFECTION BY DNA-DNA HYBRIDIZATION

A rapid detection method has been established to identify human cytomegalovirus (HCMV) in specimens of urine and white blood cells with use of DNA-DNA hybridization technology. Since radioactive labels should be avoided if possible, the DNA-DNA hybridization procedure has also been modified to employ biotin labelled probe. Preliminary experiments for sensitivity and specificity showed that hybridization with the probe HCMV DNA EcoRI J fragment could detect 0.9 pg (~(32)P-probe) or 90 pg (biotin-probe) homologous DNA; 60 cells (~(32)P-probe) or 325 cells (biotin-probe) of HCMV infected fibroblasts, and 0.3125 TCID_(50) HCMV-AD_(169). The labelled probe failed to hybridize to DNA from ather herpesviruses, uninfected human lung fibroblasts and human placental DNA. Some specimens have been detected for HCMV by DNA hybridization with ~(32)P-probe. Of 76 urine specimens and 46 white blood cell specimens from 51 newborn infants with clinically suspected cogenital HCMV infection, 11 (12%) and 9 (19%) were positive for HCMV DNA, respectively. Of 11 white blood cell specimens from normal women of childbearing age and 47 white blood cell specimens from blood donors, 6 (55%) and 30 (63%) were positive for HCMV DNA, respectively.     

Early diagnosis and monitoring of active HCMV infection in children with syste miclupus erythematosus

Objective To investigate the prevalence and features of active human cytomegalovirus (HCMV ) infection in children with systemic lupus erythematosus (SLE) and evaluate the diagnostic value of the HCMV using antigenemia assay, serum polymerase chain re action (PCR) and serology test. Methods Twenty- one SLE children undergoing immunosuppressive therapy were enrolled in t his study. Immunofluorescence assay, PCR and serology tests were used to deter mine HCMV pp65 and p72 antigens in leukocytes, HCMV DNA in sera, and HCMV specif ic IgM and IgG antibodies, respectively. As a control group, twenty- one immuno competent children with skeletal malformation were involved in this study. Stat istical analysis was performed using Chi- square test or Fisher’s exact test (Sy stat, USA), P values less than 0.05 were considered significant. Results Active HCMV infection was diagnosed in 28.6% (6/21) of SLE patients, with none in the control group; the difference between the two groups was significant (P =0.027). Two out of 6 SLE patients developed active HCMV infection before im munosuppressive therapy and the remaining 4 patients developed SLE after immunos uppressive therapy. Among the 21 SLE children, HCMV pp65 antigenemia was detect ed in 5 patients, p72 antigenemia in 3 patients, serum HCMV DNA in 9 patients, s erum HCMV- specific IgM in 2 patients, and IgG in 19 patients. The sensitivity and specificity for diagnosis of active HCMV infection were 83.3% and 100%, res pectively for pp65 antigenemia; 50% and 100% for p72 antigenemia; 100% and 80% f or serum PCR; 33.3% and 100% for HCMV IgM serology; 50% and 100% for HCMV IgG s erology. Conclusions Compared with the control group, active HCMV infection is much more frequent in SLE children, and can occur before treatment with immunosuppressive agents, b ut most often occur after immunosuppressive therapy. In comparison with the other techniques used in this study, the pp65 antigenemia assay seems to be a b etter method for the early diagnosis and monitoring of active HCMV infection in children with SLE.