聚合酶链反应检测新生儿尿巨细胞病毒DNA

采用聚合酶链反应技术（PCR），对45例疑为巨细胞病毒（HCMV）感染的新生儿，测定其尿中的HCMV－DNA，同时与尿培养法进行比较．结果前者阳性为44．44％，后者为17．78％（P＜0．01）。从而证实PCR法具敏感、特异、简便、可靠等优点，达到对HCMV感染的早期诊断和及早干预的目的。     

利用聚合酶链反应技术（PCR）测定早孕绒毛组织中巨细胞病毒DNA的研究

以人工合成的人巨细胞病毒DNA片断为引物,利用聚合酶链反应技术(PCR)检查早孕绒毛标本中人巨细胞病毒感染,对50例人工流产的早孕绒毛组织标本进行PCR基因扩增,40例绒毛标本中捡出HCMV。与同一绒毛标本的免疫组织化学染色结果相比较,PCR技术为早期、快速进行产前诊断,决定处理原则提供了一个新的检测手段。     

聚合酶链反应改良法检测石蜡包埋组织中的巨细胞病毒

聚合酶链反应改良法检测石蜡包埋组织中的巨细胞病毒钱丽清，张忠德，程新建，梁书锋，吴圣楣，顾友梅我们用二甲苯脱蜡粗提ＤＮＡ，简化了ＤＮＡ抽提步骤，用二次扩增代替一次扩增，提高了检测的敏感性，探求到一种既简便又敏感的用于石蜡包埋组织检测的改良聚合酶链反应...     

人巨细胞病毒包涵体--病毒核酸和特异阶段表达抗原的产物

目的 探讨人巨细胞病毒包涵体的组成及意义。方法 采用显微切割技术结合PCR和Southern杂交及免疫组织化学的催化信号扩增法，检测人巨细胞病毒的核酸和3种不同阶段表达的抗原。结果 通过液压显微切割系统将组织中巨细胞病毒包涵体游离出来，PCR扩增出预期长度的DNA片段，Southern杂交证实为HCMV的核酸片段，催化信号扩增法检测结果显示包涵体主要表达立即早期和早期抗原DDG9／CCH2，而基质蛋白AACl0则为阴性。结论 人巨细胞病毒包涵体的成分包含有病毒DNA和特异阶段表达的抗原产物。     

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.     

聚合酶链反应检测早孕妇女巨细胞病毒感染的临床研究

本文应用聚合酶链反应（ＰＣＲ）对１０６例早孕妇女进行人巨细胞病毒（ＨＣＭＶ）检测。其中血标本４９例,阳性者５例,阳性率１０．２％,宫颈分泌物标本５７例,阳性者７例,阳性率１２．３％,总阳性率１１．３％,并对１２例ＨＣＭＶ感染孕妇所分娩新生儿的血液标本进行了检测,其中有５例为阳性,宫内传播率４１．７％,表明ＰＣＲ法具有快速,简便,高度的敏感性和特异性的特点,是早期,快速产前诊断ＨＣＭＶ感染的最佳技术     

用逆转录套式PCR检测风疹病毒RNA

目的：介绍一种简便的快速准确检测标本中风疹病毒ＲＮＡ的分子撑方法髟于诊断孕妇感染和胎和的先天性感染风前病毒。方法：选择风疹病毒基因组中编码膜糖蛋白Ｅ１基因中的一个片段作为靶序列进行Ｂ Ｒｅｓｔｅｄ－ＰＣＲ扩增，扩增片段的长度用Ｋｏｄａｋ数码凝胶分析软件１Ｄ２．０３版本进行分析，并用内切酶酶切进行证实。结果：风疹病毒在早孕妇女中的阳性率为１．６３％；在中妊娠羊水中的阳性率为１．１７％。在畸形的儿或死     

用套式PCR检测孕妇宫颈粘液巨细胞病毒

本文用套式ＰＣＲ检测孕妇宫颈粘液中巨细胞病毒，阳性率为７８．５％（８０／１０２）此套式ＰＣＲ灵敏快速，可应用于临床检测孕妇期巨细胞病毒感染。     

原位PCR技术检测石蜡包埋脑组织中人巨细胞病毒DNA

应用原位聚合酶链反应（ＩＳＰＣＲ）技术检测了２５例尸检畸形胎儿石蜡包埋脑组织中人巨细胞病毒（ＨＣＭＶ）ＤＮＡ，并与普通ＰＣＲ及原位杂交（ＩＳＨ）进行了比较。ＩＳＰＣＲ、ＰＣＲ及ＩＳＨ检测阳性率分别为４４％，３６％及２０％。与ＩＳＨ相比较，ＩＳＰＣＲ不仅检出阳性率高，而且信号强度增强。研究结果提示，ＩＳ－ＰＣＲ是诊断ＨＣＭＶ感染的快速、敏感、特异的实用方法。     

聚合酶链反应-反向杂交快速检测巨细胞病毒糖蛋白gB、gH基因型变异

目的 建立一种简便、快速、敏感和特异的检测巨细胞病毒（HCMV）糖蛋白gB、gH基因型变异的方法。方法 以HCMV糖蛋白基因gH、gBn、gBclv为靶序列，设计17条具有型特异性的寡核苷酸探针，聚合酶链反应(PCR)扩增目的片段，反向杂交检测23例中国和6例德国移植患者HCMVgH、gBn、gBclv基因型变异，其结果与直接测序进行比较。结果 23例中国患者HCMV糖蛋白为gH1和2型，gB1、2和3型，未见gB4型；而6例德国患者的HCMV糖蛋白涵盖gH1和2型，gB1、2、3及4型。中、德国两患者均可同时感染2种不同基因型的病毒株。反向杂交技术对检测患者同时感染不同基因型病毒株优于直接测序。结论 PCR－反向杂交技术具有简便、快速、敏感和特异的特点，适用于临床实验室检测HCMV糖蛋白基因型变异。     

Detection of human papillomaviruses in squamous cell carcinomas of the lung

 The aim of this study was to evaluate the possible association of human papillomaviruses (HPV) with the development of squamous cell lung carcinomas (SqCLCs). Tissue material from 52 cases of SqCLCs were studied, and the data were evaluated according to the degree of differentiation, HPV presence and type. Analysis was performed by polymerase chain reaction (PCR) method using consensus primers, and the results were confirmed by subsequent Southern blot hybridization. Overall, the results showed 69% positivity (n=32). Forty-one cases were examined for the presence of specific HPV types (6/11 and 16/18) by hybridization of the PCR products with ³²P-labelled probes. HPV 6/11 types were detected in 6 of the 29 positive cases (20.6%). HPV 16/18 types were the most prevalent types, and were detected in 11/29 cases (37.9%: 4/10 of well-differentiated cases, 6/25 of moderately and 1/6 of poorly differentiated carcinomas). Our results confirm the possibility that HPV might play a role in the development of SqCLCs and suggest a possible relation of high-risk HPV16/18 types to tumour differentiation. Received: 3 June 1997 / Accepted: 9 February 1998     

Detection of cytomegalovirus DNA in cerebrospinal fluid in immunocompetent patients as a sign of active infection

Detection of cytomegalovirus (CMV) DNA by the polymerase chain reaction (PCR) in samples of cerebrospinal fluid (CSF) has been shown to be a sensitive method of diagnosing CMV disease in the central nervous system. Since CMV causes latent infection in white blood cells, an unanswered question is whether detection of latent CMV DNA in the cell fraction of CSF samples by PCR is possible in seropositive patients. In a prospective study, the finding of CMV DNA in CSF of CMV seropositive patients with suspected viral infection of the central nervous system (CNS) was evaluated clinically. Fractionation of 64 CSF samples from seropositive patients was carried out before analysing the samples for CMV DNA by PCR. In four of the five patients who had CMV DNA in the cell pellet and/or supernatant, the clinical data suggested CMV-associated neurological disease. The remaining 59 samples were negative in both pellet and supernatant. In addition, 11 CSF samples with high cell counts from patients with bacterial meningitis were examined for CMV DNA and found to be negative in 10 patients and positive in 1. One hundred thirty two uncentrifuged CSF samples were used as negative controls. The results of the study indicate that detection of CMV DNA in CSF samples by PCR correlated well with disease and was not due to latent CMV infection. © 1995 Wiley-Liss, Inc.     

新生儿肺透明膜病与先天性人巨细胞病毒感染的相 …

目的 探讨新生儿肺透明膜病与先天性人巨细胞病毒（ＨＣＭＶ）感染之间的关系。方法 选择ＨＣＭＶ直接早期保守区基因序列，合成两对引物，应用套式聚合酶链式反应检测了３０例新生儿肺透明膜病患儿和１０例新生儿非肺透明膜病患儿的肺组织石腊组织石腊切片中的ＨＣＭＶ ＤＮＡ。结果 ３０例新生儿肺透明膜病患儿肺组织石蜡切片标本中有１１例（ＨＣＭＶ ＤＮＡ阳性，阳性率为３６．７％，对照组肺组织ＨＣＭＶ ＤＮＡ 阴性。     

荧光原位杂交技术在未培养羊水快速产前诊断中的应用

目的建立运用荧光原位杂交技术（fluorescence in situ hybridization,FISH）检测未培养羊水细胞染色体数目的产前诊断方法。方法对100例妊娠15—26周抽取的羊水未经培养进行FISH检测,均采用13、21、18、X、Y号染色体荧光探针检测,同时行羊水培养染色体核型分析,比较两种检测方法结果的一致性。结果100例产前诊断者中未培养羊水细胞FISH检测均于48h内完成,发现6例21-三体和1例18-三体,与羊水培养染色体核型分析结果一致,后者显示6例21-三体中4例为完全型、2例为易位型21-三体。结论FISH技术与传统的羊水培养染色体核型分析相比较,具有方法快速、简便、准确可靠的特点,但无法完全取代传统的染色体核型分析,应两者结合应用于临床。     

通用引物PCR同时检测胎儿组织中4种疱疹病毒

目的：介绍一种敏感特异简便快速同时检测４种疱疹病毒的通用引物ＰＣＲ，用于临床检测和流行病学研究。方法：根据它们的高度同源序列－ＤＮＡ聚合酶基因序列设计至引物能同时扩增这４种病毒，根据它们扩增片断的大小就可将它们区分，无需再用内切酶消化。结果：总的阳性率：ＣＭＶ＝２０．１８％，ＨＳＶ＝１４．０７％，ＥＢＶ＝１．４８％。结论：１．本研究建立的通用引物ＰＣＲ是一种敏感特异简便快速能同时检测４种疱疹病毒的     

Detection of cytomegalovirus in human placental cells by polymerase chain reaction

Human cytomegalovirus (HCMV) is the most common cause of viral intrauterine infection. Progress in rapid, specific, and dependable detection of HCMV has recently been achieved by the use of DNA hybridization techniques and other molecular methods. We examined 21 placentas after delivery for the presence of HCMV DNA by polymerase chain reaction (PCR). To test the reliability of the PCR for the detection of HCMV DNA in clinical specimens, two simple PCR assays and a real-time quantitative PCR were used. PCR analysis of villous and decidual cells showed that HCMV DNA was present in 16 placentas (76.2%). Transmission of HCMV infection to chorionic villi was confirmed in 11 organs (52.4%), and congenital infections in newborns were detected in 9 cases (42.8%). These results suggest that HCMV genome detection in placentas at later gestational ages is common. Our results demonstrated that detection of HCMV DNA in placental tissues by DNA amplification provides a specific and sensitive method for diagnosis of intrauterine HCMV infection.     

肺癌组织中人乳头瘤病毒16和18型DNA相关序列的检测

采用聚合酶链反应技术（ＰＣＲ）及用ＰＣＲ法制备生物素标记探针的斑点杂交法,检测５０例肺癌、１８例肺良性病及４例胎肺组织中高危险型人乳头瘤病毒（ＨＰＶ）１６、１８型ＤＮＡ相关序列,以探讨ＨＰＶ与肺癌之间发生的关系。结果３２％的肺癌组织中检测出ＨＰＶ１６、１８ＤＮＡ,其中ＨＰＶ１６ＤＮＡ阳性１０例,ＨＰＶ１８ＤＮＡ阳性５例,１例同时含ＨＰＶ１６、１８ＤＮＡ。２７例鳞癌组织中ＨＰＶＤＮＡ阳性率为４８．２％。而肺良性病组织及胎肺组织均未发现ＨＰＶＤＮＡ。ＨＰＶＤＮＡ呈阳性的肺癌患者绝大多数为重度吸烟者。提示原发性肺癌的发生可能与ＨＰＶ感染有关。     

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。