抗-HCV阳性血清HCV RNA检测与基因分型的研究

目的 检测抗-HCV阳件血清巾的HCV RNA并进行HCV基因分型.方法 采用荧光定量PCR法检测85例大连地区抗-HCV阳性患者血清中HCV RNA,应用型特异性引物逆转录套式PCR法对HCV RNA阳性样本进行基因分型.结果 85例的抗-HCV阳性患者中,HCV RNA阳性65例(76.5%),其中基因分型1b型32例(49.2%),2a型29例(44.6%),未分型4例(6.2%).结论 抗-HCV阳性并非HCV直接标志,大连地区HCV基冈1b型和2a型基本相等.     

湖北地区丙型肝炎病毒基因型分布及流行病学特征

目的分析湖北地区丙型肝炎病毒(HCV)基因型分布及其流行病学特征。方法收集2013年5月至2016年3月在华中科技大学同济医学院附属同济医院接受HCV基因分型检查的丙肝患者血清及相关流行病学资料。采用实时荧光定量PCR对HCV RNA进行测定;采用巢式RT-PCR对HCV RNA阳性样本的核心区(core-e1)进行扩增并测序;对所测序列进行基因分型和进化树分析,并对各基因型相关的危险因素进行综合分析。结果共有HCV RNA阳性310例,测序223例,测序结果显示湖北地区存在5种基因型,1b型178例(79.82%),2a型32例(14.35%),6a型8例(3.59%),3b型4例(1.79%),3a型1例(0.45%)。1b、2a型主要见于1997年以前的有偿供血者和输血者;2a还多见于通过性接触传播的人群;6a、3b和3a型仅见于静脉注射毒品者。结论湖北地区HCV基因型分布呈现多样性且与传播途径密切相关,有关各方应予以关注及干预。     

母乳中的丙型肝炎病毒

丙型肝炎病毒(HCV)在母婴之间可通过多种途径传播,诸如子宫内、血液(出生时)以及唾液和母乳等,但是,通过母乳途径传播的资料尚不一致。作者对取自20例血清抗-HCV阳性妇女产后数天内的乳汁进行了研究。每例均筛检血清HCV RNA(PCR法)和转氨酶,并采用PCR测定母乳中的抗-HCV(半定量)及HCV RNA。 结果表明,血清抗-HCV比率>10O者的血液中均可查到HCV RNA,而抗体比     

544例新确证HIV阳性病例丙肝感染状况调查

目的了解新确诊的人类免疫缺陷病毒(HIV)阳性者中感染丙肝病毒(HCV)的现状。方法从"艾滋病综合防治信息系统"中查询"确认(替代策略)检测阳性日期"在2016年1月1日至12月31日,"郑州市第六人民医院"确证HIV阳性已终审的病例544例,对其进行HCV抗体检测,HCV抗体阳性者检测HCV核酸。结果544例新确证HIV阳性病例中,男性455例(占83.6%),女性89例(占16.4%),性别比是5∶1;性传播,占98.3%(535/544),其中同性传播占40.3%(219/544);抗-HCV阳性率是8.82%(48/544);HCV RNA阳性率是5.15%(28/544)。经性传播感染535例病例中,抗-HCV阳性率是8.22%(44/535),HCV RNA阳性率是4.49%(24/535)。经同性传播的219例病例中,抗-HCV阳性率是4.11%(9/219),HCV RNA阳性率是2.28%(5/219);经异性传播途径的316例病例中,抗-HCV阳性率是11.08%(35/316),HCV RNA阳性率是6.01%(19/316)。结论新确诊的艾滋病病例中,丙肝阳性率较高,高于普通就诊人群。在艾滋病高危人群中宣传艾滋病防治知识,同步宣传丙肝的防治知识。     

沈阳地区不同人群HCV感染及HCV基因型分布研究

应用酶联免疫检测技术 ,检测沈阳地区 390 2例不同人群血清的抗 - HCV,并应用引物特异 PCR法对其中 10 0例血清标本进行 HCV基因分型。结果 :正常人群抗 - HCV阳性率为 0 .4 2 %～ 1.66% ;输血后及散发性肝炎 (除外 HAV、HBV、EBV、CMV感染 )、乙型肝炎、肝炎后肝硬化 (除外 HBV感染 )、原发性肝癌等组抗 - HCV阳性率显著高于无输血史的非肝病病人组 ( P<0 .0 1) ;10 0例血清标本中 HCV- 型感染占 58%、HCV- 型 2 7%、 / 混合型 14%、未分型 1% ,正常人群HCV感染者中 HCV基因型以 型为主 ( 80 % ) ,丙型肝炎后肝硬化病人 HCV基因型以 型为主( 91.7% )。提示 :各类肝病病人是 HCV感染的高危人群 ;沈阳地区 HCV感染以 型为主 ,其次为 型及 / 混合型 ,HCV基因型可能与丙型肝炎病情轻重有关。     

聚合酶链反应检测血浆及外周血单个核细胞中HCV-RNA的意义

目的　为提高丙型肝炎病毒 RNA（HCV- RNA）的检出率。方法　采用套式逆转录聚合酶链反应 （RT- PCR）技术 ,检测 2 0 0例抗 - HCV阴性 ,AL T反复异常 ,临床疑似丙型肝炎患者的血浆及外周血单个核细胞 （PBMC）中 HCV- RNA。结果　 5 2例患者血浆中检出 HCV - RNA,阳性率为2 6 % ,78例患者 PBMC中检出 HCV- RNA ,阳性率为 39% ,其中两者同时双阳性有 2 2例 ,阳性率为 11%。结论　同步检测丙型肝炎患者血浆及 PBMC中 HCV- RNA,在避免漏诊 ,提高临床诊断率及指导医生治疗方面是很有必要的 ,同时在研究其发病机理中有着一定意义。     

福州市美沙酮替代治疗人群丙肝、乙肝、艾滋病和梅毒感染调查

目的了解美沙酮替代治疗者丙肝、乙肝、艾滋病及梅毒感染情况及高危因素,为制定防控措施提供依据。方法对接受美沙酮替代治疗的1 149例进行问卷调查;采集静脉血标本,用ELISA法检测抗-HCV、HBV标志物、抗-HIV和梅毒抗体。结果 1 149例中男性检出抗-HCV 569例(49.5%),HBsAg阳性25例(2.2%),抗HIV阳性31例(2.7%),梅毒抗体阳性5例(0.4%)。抗-HCV阳性者中,女性感染率(63.4%)高于男性(46.8%),感染率最高为30~49岁组。注射吸毒为HCV感染的高危因素。93例HCV感染者中,HCV RNA大于2.7log10拷贝/mL有61例(65.6%)。61例HCV RNA阳性标本中,基因型以1b型为主(65.7%)。结论美沙酮替代治疗的吸毒人群中,抗-HCV阳性率远超自然人群,尤以注射吸毒者感染率为高;其HCV基因型与福建自然人群感染丙肝基因型一致;65.6%HCV感染者存在HCV RNA高复制,需要进行抗病毒治疗。     

青岛地区丙型肝炎病毒基因分型

对青岛地区86例抗HCV阳性肝炎患者血清,采用逆转录-套式基因扩增法(RT-PCR)检测HCV·RNA,并用多引物扩增进行基因分型.结果,HCV·RNA阳性50例(58.1%),其中HCVⅡ型45例(90.0%)、Ⅲ型3例、Ⅱ/Ⅲ混合型2例.说明青岛地区HCV基因以Ⅱ型为主,应据此设计预防和治疗措施.     

湖南省美沙酮门诊就诊者感染丙型肝炎病毒基因型特征分析

目的 了解湖南省美沙酮门诊就诊者感染丙型肝炎病毒(HCV)基因型特征。 方法 收集湖南省内14个市州18岁以上、未接受抗病毒治疗、既往HCV抗体阳性的美沙酮门诊患者的血浆样本,每市州样本15份,共210份。对收集到的血浆样本分别进行HCV抗体ELISA复检、荧光PCR法定量HCV-RNA检测和基因分型检测。 结果 在210名调查对象中,ELISA法复检HCV抗体阳性标本202例,阳性率96.19%;荧光PCR法检出HCV- RNA病毒载量>15 IU/ml标本181例,阳性率86.19%。HCV基因型分别为1型18例(9.94%),2/3型51例(28.18%),1型混合2/3型47例(25.97%),其他型别65例(35.91%)。区域分布以非1型为主,2/3型普遍分布。 结论 湖南省HCV基因型比较复杂,有待深入研究。     

丙型肝炎病毒基因分型与肝病程度关系的研究

目的探索丙型肝炎病毒（HCV）基因型在辽宁省大连地区的分布及流行的优势型,并探讨HCV基因型与肝病程度的关系。方法采用型特异性引物逆转录巢式PCR法和限制性片段长度多态性分析（RFLP）法对85例抗-HCV阳性的慢性丙型肝炎、丙肝肝硬化、丙肝相关肝癌病人进行HCV的基因分型。结果85份抗HCV阳性标本中,HCVRNA检出率为61例,占71．76％（61／85）。在85份抗HCV阳性血清中,仅检出了61例1b和2a型感染者,其中1b型32例,占52％（32／61）;2a型29例,占48％（29／61）。结论大连地区基因型为1b型和2a型两种型,符合中国丙肝病毒基因分型,大连地区1b型和2a型基本相等。未发现1b和2a基因型因肝病严重程度而有所不同。     

Sexual and non-sexual intrafamilial spread of hepatitis C virus: Intrafamilial transmission of HCV

The rate of intrafamilial transmission of the hepatitis C virus (HCV) was investigated in 90 family members of 41 index patients with type C chronic liver disease. Antibody to HCV (anti-HCV) was detected by the EIA method (Abbott-Axsym Sys) and Hepatitis C virus RNA, by the polymerase chain reaction (Nested PCR). We also investigated the presence of anti-HCV in 350 healthy persons (control group). The subjects in the study included 38 spouses, 45 children and others (1 relative and 6 parents). Four family members including 3/38 (7.8%) spouses and 1 sister were found to be positive for anti HCV antibodies but none had HCV RNA. Anti-HCV was not detected in the children of index patients. The prevalence of anti-HCV in the families of index patients was significantly higher than in the controls (4.3% versus 0.85%) (p = 0.0355). We concluded that the intrafamilial transmission of HCV is possible but occurs at a low rate.     

用芯片研究丙型肝炎病毒不同功能区抗体

目的 用蛋白质芯片研究HCV不同功能区抗体及临床意义。方法 将抗原性强的C、NS3、NS4、NS5片段用芯片点样仪定量点加在醛基化玻片上来制作蛋白芯片。用进口HCV（ELISA）筛选出65例抗－HCV阳性标本,阳性标本用芯片检测不同功能区抗体,并测定HCV－RNA;同时用芯片检测24例抗－HCV阴性的正常献血员血清。结果 24例抗－HCV阴性标本用芯片检测均阴性;65例抗－HCV阳性标本经芯片均有1－4中抗体阳性,其中抗－C＋抗－NS3＋抗－NS54＋抗－NS5阳性率最高,占33．8％;抗－C、抗－NS3、抗NS5、抗NS4总的检出率分别为98．5％、89．2％、52．3％和50．8％;未发现单独含抗－NS3、抗－NS4的血清;抗－C＋抗NS3＋抗－NS4＋抗－NS5阳性血清中HCV－RNA检出率最高,占77．3％;HCV－RNA阳性血清中抗－NS5阳性率最高,占61．8％,说明抗－NS5与HCV－RNA最有相关性（P＜0．05）。结论 蛋白质芯片显示较高的敏感性与特异性;抗－C、抗－NS4在诊断抗－HCV都很重要,抗－NS4及抗－NS5有诊断的互补作用,而抗－NVS5在一定程度上能反映HCV的复制情况。     

HCV单克隆抗体的制备及其应用的初步研究

目的:丙型肝炎病毒(Hepatitis C Virus,HCV)感染已成为全球公认的健康难题,目前并没有很好的疫苗和特效药,尽早检出HCV感染者是实现丙型肝炎早期诊断、阻断传播的重要途径。方法:研究通过免疫Core-NS4B融合重组蛋白,制备抗HCV单克隆抗体,并获得5H5和4E7-HRP最优的抗体配对组合,检测120例血清,同时与HCV-RNA检测做比较。结果:阴性血清均未测出阳性,HCV阳性标测出23例阳性,检出率77%,HCV-RNA测出26例,检出率87%;HCV可疑标本测出15例阳性,HCV-RNA测出17例;单项ALT增高的标本测出2例阳性,HCV-RNA测出1例。结论:研究所获得的抗体能够用于HCV抗原检测,具有巨大的潜力,为建立HCV抗原检测试剂盒奠定了基础,为临床检测提供了实验依据。     

Sexual transmission of hepatitis C virus: the Turin study.

BACKGROUND: Hepatitis C virus infection (HCV) is a major cause of chronic liver disease with the risk of evolution towards hepatic cirrhosis and hepatocellular carcinoma. Numerous studies have documented the possibility of HCV transmission through blood transfusions during surgery or during intravenous drug use. The percentage of the risk of sexual transmission, in the general population not presenting the aforesaid risks is still very controversial. The aim of this study was to evaluate the concomitant levels of seropositivity for anti-HCV, in the partners of patients with chronic HCV liver disease, but no history of previous transfusions or use of intravenous drugs. METHODS: The study included 196 anti-HCV positive spouses with a clinical diagnosis of active chronic hepatitis, aged between 20 and 75 years (mean age 53 years, SD+/-11 years). HCV infection was diagnosed by positivity of serum samples for anti HCV (EIA), confirmed by RIBA II and by circulating HCV-RNA detected by polymerase chain reaction (PCR). All partners underwent anti-HCV assay (EIA), confirmed by RIBA II in the event of positivity. RESULTS: The mean period of cohabitation was 27 years (range 3-37, SD+/-9.8 years). The positivity of anti HCV in both subjects affected 11 couples (5.6%). Of these couples, the viral genotype was also available in 3 cases which proved to be identical in the index patient and the partner, whereas it was not possible to identify the genotype in other couples owing to scarce compliance. CONCLUSIONS: The data obtained from this study confirm the possibility of the sexual transmission of HCV. However, in the context of subjects not belonging to "high risk" groups, this method of transmission does not appear to be important if compared with that of other viruses (HBV and HIV).     

抗—HCV阳性血引发感染的核酸证据

血液和血液制品是丙型肝炎的主要传播方式之一,供血者健康检查标准规定凡ＥＬＩＳＡ检测抗ＨＣＶ抗体阳性者均不得献血。但ＥＬＩＳＡ检测存在着漏检及假阳性问题,并且只有ＨＣＶ－ＲＮＡ阳性血清才具有感染性。因此,我们对某血站送检的１００例抗ＨＣＶ阳性献血员的血样进行了ＨＣＶ－ＲＮＡ检测,结果表明,在ＨＣＶ－ＲＮＡ半套式ＲＴ－ＰＣＲ中,传统的异硫氰酸胍／氯仿／异戌醇方法同磁珠法抽提病毒ＲＮＡ,结果两种方法同样可以得到理想的２４０ｂｐ扩增带。用传统方法进行实验,１００例血标本中３例ＨＣＶ－ＲＮＡ阴性,其它均阳性,说明我国制定的献血员标准规定绝大多数情况下是适宜的,如果结合ＲＴ－ＰＣＲ检测ＨＣＶ－ＲＮＡ,结果会更加理想。这为库血ＨＣＶ检测与献血员ＨＣＶ感染检测提供了一种可靠的检测方法,对防止输血后肝炎的发生、净化献血员有一定的指导意义     

Hepatitis C virus transmission in family members of Egyptian patients with HCV related chronic liver disease

Hepatitis C virus (HCV) infection in Egypt has become a major public health problem. In the present study, sexual and intrafamilial transmission of HCV infection were assessed in the family members of 200 Egyptian patients (index patients) with HCV-RNA positive and biopsy ascertained chronic hepatitis C. Index patients were 139 men (mean age 55+/-11 years) and 61 women (mean age 48+/-8 years). Family members consisted of 200 spouses; 139 women (mean age 45+/-12 years) and 61 men (mean age 58+/-9 years); and 355 children (183 males and 172 females, mean age 11.8+/-10 years). All the family members were tested for the presence of antibodies to HCV in their sera. Thereafter, HCV-RNA detection by PCR and HCV serotype determination were performed in antibody positive contacts. Hepatitis C virus antibodies were detected in 28 (14%) spouses, all of them were also positive for HCV-RNA. Hepatitis C virus serotypes were identical in HCV seropositive patient-spouse pairs (Serotype 4). None of the 355 children involved in this study showed HCV antibodies in their sera. No significant difference was found between the prevalence of male-to-female and female-to-male transmission of HCV. A highly significant association was found between both the age of the spouse and the duration of marriage to index patient and HCV seropositivity in spouses. Moreover, HCV seropositivity in spouses was significantly related to increased serum ALT and HCV-RNA levels, histological severity of chronic hepatitis C and to a history of dental care, as a risk factor for HCV acquisition, in index patients. It was concluded that spouses of patients with HCV viremia and chronic liver disease have an increased risk for acquiring HCV, while intrafamilial acquisition of HCV in non sexual contacts seems to be rare. The authors suggest that spouses of HCV viraemic patients should be followed routinely for markers of HCV infection and liver disease.     

Die chronische Hepatitis C der Mutter ist keine Kontraindikation für das Stillen

To evaluate the risk of hepatitis C virus (HCV) transmission via breast milk, we collected 165 samples of breast milk from 143 chronically HCV-infected women at 152 deliveries (153 children). Samples were collected between the 31st day prior to and the 384th day after delivery (median, fourth day post partum). None of the 165 milk samples were positive for HCV RNA. To exclude inhibitory effects of components of the milk on our PCR, 120 milk samples were divided and spiked with both serum drawn from an HCV negative and an HCV infected patient. In 79 of 99 samples spiked with serum drawn from a viremic patient, HCV-RNA could be detected. Thus, milk does not interfere with the PCR. In 134 cases, a maternal serum sample was drawn during pregnancy or until 6 months thereafter (median, 3 days post partum) and tested for HCV virus. Of these, 89 women (66.4%) had positive PCR results with concentrations of HCV RNA between 1×10² and 1×10⁷ copies/ml (median, 6×10³ copies/ml). Of 153 breastfed children, 151 could also be tested for HCV. Only three of these (2.0%) had acquired the infection; a rate which is less than the 4.0% observed in the collective of 624 vertically HCV exposed children studied at our center. Two of these HCV positive children had already become infected in utero and one of them probably cleared the virus. The third HCV positive child obviously became infected late in pregnancy or around delivery. Hence, chronic HCV infection of a woman is no contraindication for breast-feeding.     

丙型肝炎病毒窗口期感染核酸阳性血清检测和基因型别分析

目的分析丙型肝炎病毒（HCV）早期感染血液样品核酸（RNA）检测的意义，并对检出的核酸阳性血清进行基因分型。方法对8649份自然供血员及566份HCV可疑感染者血清进行核酸（TMA RNA试剂）检测，检测出的HCV RNA阳性样品利用基因分型芯片进行分型。同时利用HCV核心总抗原（HCVc Ag）和游离抗原（HCV free Ag）试剂以及抗体（Anti—HCV）试剂进行检测。并对检测数据进行比较分析。结果在所有血液样品中发现1份HCV抗体、HCVc Ag、HCV free Ag全部阴性但RNA阳性的样品，基因型别为1b型。在8649份自然供血员血清中筛出1份HCVc Ag、HCV free Ag以及HCVRNA阳性但抗体阴性血清，基因型别为1b型。在566份HCV可疑感染者血清中，筛出1份HCV free Ag和核心抗体同时阳性血清，基因型别为1b型。TMA试剂筛出5份RNA阳性血清但微粒子抗体试剂检测阴性，TMA试剂筛出6份RNA阳性的血清而酶标抗体试剂检测阴性。这6份血清，利用傲拓HCV总抗原试剂检测，其中5份HCV总抗原阳性。在澳大利亚国家血液中心进行复核，其中5份RNA阳性。在566份血清中，TMA试剂检出RNA阳性495份，阳性率87．4％。对RNA阳性血清进行型别分析，以1b、2a型为主，分别为46．5％和27％。结论由于检出了HCV RNA单独阳性的窗口期样品，认为利用HCV抗体试剂对血液进行筛查的同时，使用HCV RNA试剂做必要的补充实验，可以减少因窗口期造成的漏检。型别分析认为，窗口期感染样品基因型别也以1b，2a型为主。     

腹膜透析患者HCV感染标志检测及临床意义

目的：评价维持性腹膜透析（腹透）患丙丙型肝炎病毒（HCV0感染状况及其临床意义。方法：采用第二代酶链免疫吸附试验（ELISA2）和逆转录聚合酶链反应（RT－PCR）检测60例腹透患者血清抗－HCV和HCVRNA,按HCV标志阳性与阴性分组比较有关临床因素。结果：60例患者中,抗－HVCIgM阳性7例（11．7％）,抗－HCVIgG阳性10例（16．7％）,HCVRNA阳性13例（21．7％）,三项中任一项阳性15例（25．0％）;阳笥组与阴性组相比,两组在性别、年龄、肾功能、HBV标志、EPO（促红细胞生成素）应用上无明显差异,而在平均透时间、输血史、ALT异常、血液透析史、肾移植史方面有显著性差异。结论：腹透患者中HCV感染较为严重,输血为腹透患者感染HCDV的主要途径,但也存在与透析环境有关的传播的可能。