应用PCR法检测新生儿肝炎综合征患者HCMV－DNA

应用ＰＣＲ法检测新生儿肝炎综合征患者ＨＣＭＶ－ＤＮＡ长春市儿童医院应用ＰＣＲ检测３０例临床诊断为新生儿肝炎综合征（ＮＨＳ）患者和１４例正常新生儿尿中人巨细胞病毒ＤＮＡ（ＨＣＭＶ－ＤＮＡ）。结果阳性率分别为６６．７％和１４．２９％，两者差异显著。ＨＣＭ...     

应用聚合酶链反应检测新生儿黄疸患儿巨细胞病毒感染

本文对排除了引起新生儿黄疸常见病因的３９例患儿，用聚合酶链反应检测患儿外周血人巨细胞病毒ＤＮＡ。其中ＨＣＭＶ ＤＮＡ阳性９例，阴性３０例，对两组黄疸患儿进行比较，ＨＣＭＶ ＤＮＡ阳性组其血清总胆红素水平及黄疸消退时间与阴性组比较明显显著差异，两组血小板计数无差异。     

巨细胞病毒宫内感染的诊断及对胎儿的影响

采用地高辛探针斑点杂交技术和聚合酶链反应（ＰＣＲ）技术，对５１例人巨细胞病毒（ＨＣＭＶ）血清学抗体ＩｇＭ阳性孕妇的羊水、脐血和产后两周内的新生儿尿液，进行ＨＣＭＶＤＮＡ检测。同时与酶联免疫吸附法（ＥＬＩＳＡ）检测脐血、羊水中ＨＣＭＶ－ＩｇＭ的结果相比较。结果，地高辛探针斑点杂交检测ＨＣＭＶ－ＤＮＡ的阳性率分别为：羊水２１．５７％，脐血３３．３３％，新生儿尿液２７．４５％。ＰＣＲ检测ＨＣＭＶ－ＤＮＡ的阳性率分别为：羊水２９．４１％，脐血４３．１４％，新生儿尿液３Ｓ，２９％。羊水ＨＣＭＶ－ＩｇＭ阳性率为１１．７６％，脐血ＨＣＭＶ－ＩｇＭ阳性率为２３．５３％。脐血ＨＣＭＶ－ＤＮＡ阳性的新生儿平均出生体重明显低于脐血ＨＣＭＶ－ＤＮＡ阴性的新生儿，而脐血ＨＤＷ－ＤＮＡ阳性的新生儿肝功能异常、血小板减少以及低Ａｐｇａｒ评分的发生率明显高于脐血ＨＣＭＶ－ＤＮＡ阴性的新生儿。结果表明：采用ＥＲ技术检测羊水、脐血或产后两周内新生儿尿液ＨＣＭＶＤＮＡ有助于ＨＣＭＶ官内感染的早期诊断，便于临床早期干预，ＨＣＭＶ官内感染严重影响胎儿的生长发育，可造成胎儿肝功能异常、血小板减少和新生儿窒息的发生。     

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。     

用聚合酶链反应诊断新生儿巨细胞病毒感染25例分析

本文采用聚合酶链反应技术（ＰＣＲ），对２５例婴儿肝炎综合征患儿尿中ＨＣＭＶ－ＤＮＡ，用于确诊新生儿巨细胞病毒感染，阳性率为６８％。占本组病毒感染的首位。在新生儿黄疸发生率较高的广西，对原因不明的婴儿肝炎综合征患儿，应争取做患儿尿中ＨＣＭＶ－ＤＮＡ，因此法具有高度特异性、敏感性，取材方便，操作简便，反应周期短等优点，是目前检测ＨＣＭＶ的最敏感的实验方法，适用于临床推广使用。     

急性白血病患儿脑脊液PCR扩增HCMV－DNA的研究

应用多聚酶链式反应（ＰＣＲ）扩增和洋地黄探针杂交的方法，对５３例急性白血病（ＡＬ）患儿脑脊液（ＣＳＦ）样本进行了检测。经过ＰＣＲ扩增，可见明显的人巨细胞病毒ＤＮＡ（ＨＣＭＶ－ＤＮＡ）４００ｂｐ扩增带；经洋地黄探针特异性杂交鉴定，ＡＬ患儿ＣＳＦ标本ＰＣＲ扩增产物的ＨＣＭＶ－ＤＮＡ阳性检出率为８３．０２％。本实验为ＡＬ病人ＣＳＦ的ＨＣＭＶ检测提供了一个简便方法。     

急性白血病患儿脑脊液PCR扩增HCMV—DNA的研究

应用多聚酶链式反应（ＰＣＲ）扩增和洋地黄深针杂交的方法，对５３例急性白血病（ＡＬ）患儿脑脊液（ＣＳＦ）样本进行了检测，经过ＰＣＲ扩增，可见明显的人巨细胞病毒ＤＮＡ（ＨＣＭＶ－ＤＮＡ）４００ｂｐ扩增带，经洋地黄探针特异性杂交鉴定，ＡＬ患儿ＣＳＦ标本ＰＣＲ扩增产生的ＨＣＭＶ－ＤＮＡ阳性检出率为８３．０２％。本实验为ＡＬ病人ＣＳＦ的ＨＣＭＶ检测提供了一个简便方法。     

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...     

用PCR技术产前诊断人巨细胞病毒感染

用ＥＬＩＳＡ法筛选早孕妇女血人巨细胞病毒ＩｇＭ（ＨＣＭＶ－ＩｇＭ），在孕１２￣２０周用聚合酶链式反应（ＰＣＲ）技术检测其羊水中ＨＣＭＶ－ＤＮＡ，对胎儿巨细胞病毒感染作出产前诊断。在３８４例孕妇中，筛选出血ＨＣＭＶ－ＩｇＭ阳性２６例为实验组，其羊水ＨＣＭＶ－ＤＮＡ检出率为３４．６２％；从２５８例ＨＣＭＶ－ＩｇＭ阴性中选出２０例为对照组，其羊水ＨＣＭＶ－ＤＮＡ检出率为１０．０％，两组比较有显著性差异（     

石家庄地区新生儿尿巨细胞病毒感染的临床分析

采用聚合的酶链反应（ＰＣＲ）检测１０５例出生后３日内新生儿尿中巨细胞病毒,同时应用酶联免疫吸附法（ＥＬＩＳＡ）检测脐血特异性ＩｇＧ和ＩｇＭ抗体。结果,新生儿尿液ＨＣＭＶ－ＤＮＡ阳性者３３例,阳性率３１．４％,脐血ＩｇＧ抗体阳性率１００％,ＩｇＭ挤体阳性率１．９％。结果表明ＰＣＲ技术检测新生儿尿液ＨＣＭＶ－ＤＮＡ有利于ＨＣＭＶ先天性感染的早期诊断,早预防、早阻断。有利于更好地实行计划生育,有利于优生     

早期妊娠胚胎先天性人巨细胞病毒感染检测及危险因素分析

本文利用免疫组织化学方法(PAP)和ELISA法对400名早期妊娠胚胎绒毛组织进行人巨细胞病毒(HCMV)先天性感染和孕妇活动性HCMV抗体检测。结果表明早孕绒毛中有67例感染，阳性率为16．8％；孕妇血中HCMV—IgM( )22例，HCMV—IgA( )27例，孕妇活动性HCMV感染率为10、0%。并对早期妊娠胚胎感染HCMV危险因素进行分析，孕妇活动性HCMV感染传播给胚胎的危险度为32.5％．而且胚胎先天性感染与妊娠妇女的妊娠天数和性生活频度有着密切的相关关系。证明孕早期胚胎HCMV感染既可由母亲活动性HCMV感染垂直传播，也可来源父亲的传染途径。     

Building a mouse model hallmarking the congenital human cytomegalovirus infection in central nervous system

To investigate the mechanisms that human cytomegalovirus (HCMV) can vertically transmit from the placenta of mice to infect their offspring in the central nervous system (CNS) and cause congenital anomalies, and in order to provide basic research for preparing HCMV vaccine, we have developed a new type of mouse model of HCMV congenital CNS infection. Pure strain mice were propagated after being infected with HCMV. Then the degree of infection by HCMV to offspring was determined. The experiment shows that in the infection groups the mortality of fetal mice and the fatality of neonatal mice in one week are higher than that of the control groups (P < or = 0.05). At the same time we investigated the CNS of fetus's mice whose mothers were infected by HCMV. Our results showed: 1. The virus was successfully isolated from their cerebral cortex. 2. The signal of HCMV hybridization print was found in their nervous cell through in situ hybridization. 3. Especially human herpes virus-like particles and inclusion bodies in the plasm of nerve cell were found in the tissue of their brain under the electron microscope. This new type of mouse model of HCMV inherent CNS infection will help prepare HCMV vaccine and research HCMV congenital infection in CNS.     

IL‐12 and type I IFN response of neonatal myeloid DC to human CMV infection

Following congenital human CMV (HCMV) infection, 15–20% of infected newborns develop severe health problems whereas infection in immunocompetent adults rarely causes illness. The immaturity of neonatal antigen presenting cells could play a pivotal role in this susceptibility. Neonatal myeloid DC were shown to be deficient in IFN‐β and IL‐12 synthesis in response to TLR triggering. We studied the response of cord and adult blood‐derived myeloid DC to HCMV infection. Neonatal and adult DC were equally susceptible to in vitro HCMV infection. Among immunomodulatory cytokines, IL‐12, IFN‐β and IFN‐λ1 were produced at lower levels by neonatal as compared with adult DC. In contrast, neonatal and adult DC produced similar levels of IFN‐α and IFN‐inducible genes. Microarray analysis indicated that among the more than thousand genes up‐ or down‐regulated by HCMV infection of myeloid DC, 88 were differently regulated between adult and neonatal DC. We conclude that neonatal and adult DC trigger a partly different response to HCMV infection. The deficient IL‐12 and mature IFN‐α production by neonatal DC exposed to HCMV are likely to influence the quality of the T lymphocyte response to HCMV infection in early life.     

HMA-SSCP方法研究人类巨细胞病毒UL144基因在临床低传代分离株中的多态性

目的 探讨人类巨细胞病毒（HCMV）UL144序列在临床患儿低传代分离株中的多态性及与临床疾病的关系。方法 对65株HCMV临床低传代分离株及7例同年龄组HCMV，DNA定量PCR方法检测阳性无症状感染儿尿液进行HCMV-ULl44 PCR扩增及HMA-SSCP分析，并对其中32份阳性标本进行测序。结果 65株分离株中有55株UL144全序列引物PCR扩增阳性，7份QPCR检测HCMV—DNA阳性无症状感染儿尿液中5份UL144全序列引物PCR扩增阳性。60份UL144扩增阳性标本HMA-SSCP（异源双链泳动及单链构象多态分析）呈现3种典型带形，巨结肠患儿分离株序列、小头畸形患儿的序列分布以1型为主，巨结肠患儿分离株序列没有2型，黄疽患儿以3型为主。结论 HCMV-UL144广泛存在于临床低传代分离株中，用HMA-SSCP检测HCMV-UL144基因在临床低传代分离株中的多态性是一种可行的方法。HCMV不同疾病类型的HCMV-UL144序列不同，提示UL144基因可能对HCMV致病性起一定作用。     

人类巨细胞病毒UL144基因在临床低传代分离株中的多态性研究

目的：探讨人类巨细胞病毒（human cytomegalovirus,HCMV)UL144序列在临床患儿低传代分离株中的多态性及其与临床疾病的关系。方法：对65株HCMV临床低传代分离及7例同年龄组HCMV－DNA定量PCR方法检测阳性健康儿尿液进行HCMV-UL144 PCR扩增分析，对HCMV-UL144扩增阳性标本进行HMA－SSCP分析，并对其中32个阳性标本进行HCMV-UL144基因全序列测定及分析。结果：65株分离株中有55株UL144全序列引物PCR扩增为阳性，7份QPCR检测HCMV-DNA阳性健康儿尿液中5份UL144全序列PCR扩增为阳性，60份UL144扩增阳性标本HMA－SSCP（heteroduplex mobility assay and single-stranded conformation polymorphism)分析呈现3种典型带形，32例个测序标本序列呈现较高的多态性，差异多位于序列的前半部，种系发生图谱分析可大致分为3组，二级结构预测表现为6种类型，在重要的蛋白质功能的序列分布以Ⅰ型为主，黄患儿以Ⅲ型为主。32个HCMV-UL144序列已被GenBank收录，结论:HCMV-UL144广泛存在于临床低传代分离株中，序列呈序高度多态性，不同疾病类型的HCMV-UL144序列不同，提示UL144基因可能在HCMV致病上起一定的作用。     

Human cytomegalovirus infection in infants with prolonged neonatal jaundice

BACKGROUND: Although human cytomegalovirus (HCMV) infection in infants has been associated with liver disease, the role of HCMV in infants presenting with prolonged neonatal jaundice is unclear as this clinical picture can be caused by a broad spectrum of underlying conditions. OBJECTIVES: This study aimed to determine a possible role for HCMV infection in infants with prolonged cholestatic neonatal jaundice that could facilitate the appropriate use of diagnostic assays and specific treatment in this condition. STUDY DESIGN: HCMV immunohistochemical staining was performed on liver biopsy specimens received for histopathological examination from 85 infants (mean age 3 months) with a clinical history of prolonged neonatal jaundice. HCMV serology was also performed. RESULTS: One infant with a histological diagnosis of HCMV hepatitis was also positive by immunohistochemical staining, while all other tissue specimens were negative for HCMV. HCMV IgG was positive in 92.3% and HCMV IgM in 39.7% of the infants (n=78). CONCLUSIONS: The serological results confirm the ubiquitous nature of HCMV with many primary infections occurring within the first year of life. Despite this, HCMV hepatitis was uncommon in this cohort.     

Congenital infection by human cytomegalovirus with a 65 bp deletion in the morphological transforming region II

Summary Human cytomegalovirus (HCMV) infection is an important cause of neonatal death. Using primers derived from sequences within the morphological transforming region II (mtrII), HCMV DNA was amplified by polymerase chain reaction (PCR) from fixed tissues of infants who had died of congenital HCMV infection. In one neonate, HCMV DNA with reduction in the expected size was detected in the liver, spleen, kidney, adrenal, und thyroid tissues by gel electrophoresis. Nucleotide sequencing of the PCR product revealed a 65bp frame-shift deletion within the 79 amino acid open reading frame (ORF) of themtrII. Based upon this observation, it is likely that viral genomic rearrangement involving themtrII may occur in some cases of congenital HCMV infection.     

高血压患者人巨细胞病毒感染率及其血浆中和抗体水平分析

目的 对高血压患者人巨细胞病毒（HCMV）感染率及其血浆中和抗体水平展开研究分析,研究高血压与人巨细胞病毒感染的相关性.方法 随机选取2011年12月-2013年12月期间接收治疗的50例高血压患者血标本作为观察组,同时选取50例健康体检人员的血标本作为对照组.对两组标本人巨细胞病毒特异性中和抗体、人巨细胞病毒特异性IgG和IgM、人巨细胞病毒特异性UL93 DNA进行检测.结果 观察组HCMV UL93 DNA的阳性率为72.0％,HCMV IgG阳性率为70.0％,HCMV IgM阳性率为4.0％;对照组HCMV UL93 DNA阳性率为54.0％,HCMV IgG阳性率为52.0％,HCMV IgM阳性率为2.0％;观察组HCMV UL93 DNA阳性率、HCMV IgG阳性率显著高于对照组,数据差异具有统计学意义（P＜0.05）;两组HCMV IgM阳性率数据对比差异无统计学意义（P＞0.05）.结论 高血压患者人巨细胞病毒感染率显著高于健康对照组,但特异性中和抗体水平较健康对照组显著下降,即高血压患者的人巨细胞病毒感染体液免疫状态不足,高血压与人巨细胞病毒感染显著相关.     

采用PCR技术对绒毛和羊水巨细胞病毒感染研究的比较分析

本文采用ＰＣＲ方法检测了早孕及孕中期羊水人巨细胞病毒的感染情况，孕早期１０１例绒毛的ＨＣＭＶ感染８７例，５４例羊水标本经ＰＣＲ检测发现ＨＣＭＶ阳性２４例，进一步分析三个孕期的ＨＣＭＶ感染表明ＨＣＭＶ可以经胎盘传播给胎儿，同时胎盘有一定的屏幕作用，而且母体胎儿自身也有相当的保护能力。此研究结果证实ＰＣＲ方法检测先天性ＨＣＭＶ感染敏感，快速，并具有特异性。     

Congenital human cytomegalovirus infection: value of human cytomegalovirus DNA quantification in amniotic fluid

A quantitative PCR assay (RS Elosa CMV, Lambdatech) was used to quantitate HCMV DNA in maternal amniotic fluid of 12 fetuses with congenital infection (group 1) and of 10 fetuses without congenital infection (group 2). HCMV detection was performed for both groups using culture and qualitative PCR. Histologic examinations of fetal tissues and placenta were carried out for 9 patients from group 1. The amniotic fluid viral loads were negative in all patients of group 2. In group 1, all viral loads were high (from 1.105 to > 107 cop/mL) and no difference was observed between symptomatic and asymptomatic foetuses. Further evaluation on larger samples is needed to define more precisely the pronostic value of HCMV DNA quantification in amniotic fluid.