丙型肝炎病毒感染患者血清肝特异性自身抗体的检测及临床研究

目的了解丙型肝炎患者血清中肝特异性自身免疫性抗体的检出率,探讨自身免疫在丙型肝炎(HCV)感染中的意义。方法采用间接免疫荧光法,对220例丙型肝炎病毒感染者血清进行肝特异性自身抗体的检测;应用聚合酶链反应定量检测血清HCV-RNA含量,HCV抗体的检测用ELISA法;回顾性分析HCV-RNA病毒、AST、ALT、ALP和r-GT与自身抗体检出的关系。结果220例丙型肝炎患者总自身抗体检出率为32.73%;HCV-IgG单阳性(HCV-IgG( )HCV-RNA(-))的检出率为29.57%;HCV-IgG( )HCV-RNA( )自身抗体检出率为40.59%,HCV-IgG( )HCV-RNA(-)与HCV-IgG( )HCV-RNA( )组比较差异有统计学意义(P<0.01)。各组与正常对照比较差异均有统计学意义(P<0.01);自身抗体以低滴度为主,主要为抗核抗体。自身阳性的检出与性别无关(P>0.05)与年龄关系密切(P<0.01)。自身抗体阳性丙型肝炎患者的血清丙氨酸氨基转移酶(ALT)、谷氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)及γ-球蛋白均高于自身抗体阴性患者,差异有统计学意义(P<0.01)。结论HCV在感染过程中可诱导自身免疫反应,使感染者体内产生多种自身抗体,自身抗体检出率与年龄关系密切且与病毒的复制有关。自身抗体以低滴度为主,主要为抗核抗体。自身抗体可能是HCV感染后肝组织损伤的重要因素,应引起重视。     

提高丙型肝炎诊断准确性的检测方法

目的 探讨避免丙型肝炎病毒感染的漏诊及提高诊断效率的检测方法. 方法 采用RT-PCR和ELISA法,检测186例疑似HCV感染患者血浆中的HCV-RNA及血清中的抗-HCV抗体. 结果 186份标本中RT-PCR检测HCV-RNA阳性标本104份,阳性率为55.91％;HCV抗体检测阳性标本116份,阳性率为62.37％,2者符合率89.66％.2种检测方法均为阳性的标本92份,阳性率49.46％. 结论 同时开展HCV-RNA与抗-HCV抗体检测可以避免HCV感染的漏诊,提高丙型肝炎诊断的准确性.     

输血前患者检测丙肝病毒核心抗原临床意义探索

[目的]应用酶联免疫法(ELISA法)进行丙肝病毒核心抗原(HCV-cAg)和抗体(HCV-Ab)检测,研究输血前患者丙肝病毒核心抗原检测的临床意义.[方法]运用酶联免疫法(ELISA)HCV核心抗原(HCV-cAg)试剂盒,有选择地对442例输血前临床患者血清标本(其中394例丙肝抗体检测阴性、48例丙肝抗体检测阳性)进行HCV-cAg检测.并对HCV-cAg阳性标本进一步作HCV-RNA检测.[结果]442例血清标本,检测出HCV核心抗原(HCV-cAg)阳性15例,阳性率3.39%,其中394例HCV-Ab阴性标本中HCV-cAg阳性2例,阳性率0.51%;48例HCV-Ab阳性的样本检出HCV-cAg阳性13例,阳性率为27.08%.[结论]丙型肝炎病毒核心抗原对丙型肝炎病毒感染的检出时间早于抗体,缩短了检测"窗口期",利于早期检测出丙肝病毒感染,避免或减少"输血后感染丙肝"等医疗纠纷.同时证实,患者感染丙肝病毒后的病程中随着抗体的出现,血液中的HCV核心抗原(HCV-cAg)大多减少或消失,因此HCV-cAg检测可作为HCV抗体检测的联合筛检方法,尤其对输血前、术前HCV筛查的患者联合应用更具有临床意义,其检测成本较PCR低廉,适合于丙型肝炎的常规临床筛查应用,对保障临床输血和手术安全起到积极作用.     

3536例健康体检者丙型肝炎病毒感染状况的调查分析

目的:了解湛江市HCV感染状况及探讨丙型肝炎感染相关因素。方法:抽取健康体检者3536例,检测丙型肝炎病毒抗体(Anti-HCV)、丙氨酸转氨酶(ALT),对HCV抗体阳性标本再作HCV-RNA检测。结果:3536份样本中HCV抗体阳性33例(9.08‰),男性26例(7.15‰),女性7例(1.92‰),男性HCV抗体检测阳性率比女性高(χ2=11.04,P<0.01);33例HCV抗体阳性标本中12例(男11例,女1例)标本HCV-RNA阳性,9例(男性8例,女性1例)标本ALT升高。结论:HCV抗体感染率不高,感染途径及易感染途径有待研究;重视丙型肝炎病毒的危害性,开展HCV抗体普查,及早发现丙型肝炎及控制感染源,切断传播途径,防止丙型肝炎病毒的蔓延。     

金标快速法筛查献血者丙型肝炎抗体的应用评价

目的:对丙型肝炎抗体(抗HCV抗体)金标快速法试剂盒进行应用评价.方法:采用ELISA法和金标快速法平行检测200份献血者、50例已确诊的丙型肝炎患者和50例健康体检者血清中的抗HCV抗体并比较结果.结果:ELISA法和金标快速法检测200份献血者血清抗HCV抗体的阳性结果分别是22例和20例,有2例ELISA法阳性而金标法阴性;检测50例已确诊的丙肝患者血清抗HCV抗体的阳性检出率分别为100%和96%,两法对50例健康体检者的结果均为阴性.以ELISA法的结果为真值,金标快速法的敏感性为947%、特异性为100%、准确性为987%.结论:金标快速法适用于抗HCV抗体的流行病学调查、单份标本和急诊检验,但其敏感度低于ELISA法,如时间允许,最好选择ELISA法进行复检.     

HCV核心抗原在丙肝检测中的应用分析

目的 HCV核心抗原在丙肝检测中的应用分析。方法本文选取80例HCV感染患者,将其血清标本作为本次研究对象,所有标本全部经过HCV核糖核酸检测显示阳性,进一步对其实施HCV核心抗原等检测过程,对其检测结果分析探讨。结果采用HCV核心抗原检测法、抗HCV抗体检测法两种检测方式开展检测工作后,发现HCV核心抗原检测法的检测准确率达95.00%,而采用抗HCV抗体检测法的检测准确率为80.00%,两种检测结果对比差异有统计学意义(P0.05)。结论 HCV感染患者采用HCV核心抗原检测方式后,对于丙肝疾病有着更高的检出率,临床应用效果较好,同时操作过程更加简单、方便,有效降低了检测成本,提升检测效果。     

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。     

丙型肝炎患者HCV-RNA感染与血清自身抗体的相关性分析

目的 研究丙型肝炎患者HCV-RNA感染与血清自身抗体产生的相关性,进一步探讨丙型肝炎的发病机理.方法 对本院消化科220例丙型肝炎抗体IgG(+)患者进行HCV-RNA检测及抗肝抗原自身抗体谱检测.结果 220例患者中,有122例HCV-RNA阳性,阳性率为55.4％,98例患者HCV-RNA阴性,阴性率为44.6％.将其分为两组,丙型肝炎抗体IgG(+),HCV-RNA阳性组,测得抗线粒体抗体M2(AMA-M2)阳性9例(7.3％),抗可溶性肝抗原抗体(SLA) 14例(11.47％),肝抗肾微粒体抗体(LKM-1)阳性26例(21.3％),抗肝细胞溶质抗原1型(LC-1)阳性7例(5.7％);丙型肝炎抗体IgG(+),HCV RNA阴性组,测得AMA M2阳性3例(2.46％),SLA阳性4例(3.28％),LKM-1阳性13例(10.65％),LC-1阳性3例(2.46％).两组肝抗原抗体谱阳性率差异有统计学意义(x2=93.4,P＜0.05).结论 丙肝患者抗肝抗原自身抗体谱检测阳性率与丙肝患者HCV-RNA阳性呈正相关.     

HCV游离抗原BA-ELISA检测方法的临床应用评价

目的根据国家相关法规对HCV游离抗原BA-ELISA检测方法进行临床应用效果评价。方法生物素标记丙型肝炎病毒抗体(HCV-Ab)与辣根过氧化物酶标记亲和素联合应用建立HCV游离抗原BA-ELISA检测法,分别在3个省级医疗卫生机构进行临床试验,同时使用HCV-cAg、HCV-Ab、HCV-RNA荧光定量检测试剂盒进行对比同步检测。结果临床试剂与HCV-Ab检测结果相比一致性66.67%,特异性100%;与荧光定量PCR相比结果一致性为80.85%、特异性为98.87%;同类市售产品HCV核心抗原检测试剂检测结果与之接近。结论临床试剂特异性较好,灵敏度有待提高;临床研究试剂比市售同类产品阳性检出率略高,但两种试剂检出率均低于核酸及抗体检测试剂。     

HCV抗原ABS-ELISA检测方法的建立及初步评价

目的 建立新的HCV抗原生物素-亲和素-酶联免疫检测方法(ABS-ELISA).方法 将生物素-亲和素系统与酶联免疫技术相结合,建立用于丙型肝炎病毒抗原检测的方法,分析检测方法的灵敏度、特异度、稳定性及精确度等主要技术指标,并对应用效果进行评价.结果 建立的检测方法,对40余种肠道病毒样品均无交叉反应,在第二代HCV-RNA国家标准品的10份阴性标准品中,未检出HCV-Ag阳性样品,在10份HCV-RNA阳性标准品中,检出6份HCV-Ag阳性的样品,表现出良好的特异性；检测试剂CV平均值为6.28％,精密性好；在90份HCV抗体阳性样品中检测出HCV-RNA样品67份(74.44％),检出HCV游离抗原阳性样品30份(33.33％)、HCV总抗原阳性样品68份(75.56％),HCV游离抗原与HCV-RNA两个指标检出率差异有统计学意义(x2=22.443,P＜0.01),但HCV总抗原与HCV-RNA检测结果之间差异无统计学意义(x2=0.037,P＞0.05)；以HCV-RNA检测试剂为金标准,所建立的ABS-HCV抗原检测试剂总抗原检测结果的一致性、灵敏度、特异度、阳性预测值及阴性预测值分别为98.03％、80.60％、98.93％、79.41％、99.00％.结论 新建立的HCV抗原ABS-ELISA检测方法特异性好,其灵敏度与荧光定量PCR相近.     

联合检测丙肝病毒核心抗原和抗体在丙型肝炎早期诊断中的价值

目的:通过检测丙型肝炎抗体(HCV-Ab),同时对其进行丙型肝炎病毒核心抗原(HCV-cAg)和丙肝病毒RNA(HCV-RNA)两项指标进行检测,探讨HCV-cAg检测在丙型肝炎早期诊断中的意义。方法:对104例HCV-Ab阳性标本及108例高危人群但HCV-Ab阴性标本(包括血液透析患者55例、医务人员20例和丙型肝炎病人的家庭密切接触者33例)及健康对照组同时进行HCV-cAg和HCV-RNA的检测。结果:104例HCV-Ab阳性组中,HCV-cAg阳性25例,HCV-RNA阳性27例;108例HCV-Ab阴性组中,HCV-cAg阳性2例,HCV-RNA阳性1例;健康对照组全阴性。结论:HCV-cAg和HCV-RNA在丙型肝炎早期诊断上无显著性差异,但HCV-cAg检测在临床的应用前景上具有更大的优势,故可在临床推广HCV-Ab和HCV-cAg联合检测,有条件者可联合检测HCV-Ab、HCV-cAg和HCV-RNA。     

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.     

丙型肝炎实验室诊断方法及相关指标的探讨

目的探讨丙型肝炎病毒RNA(HCV-RNA)定量检测及抗丙型肝炎病毒(anti-HCV)检测在丙型肝炎病毒感染诊断中的价值,以及HCV感染对血脂代谢的影响。方法收集anti-HCV阳性丙型肝炎患者与正常健康人标本分别为40和84例,检测样本血浆中HCV-RNA,血清anti-HCV抗体、超敏C-反应蛋白(hs-CRP)、载脂蛋白AI(apoAl)、载脂蛋白B(apoB)、甘油三酯(TG)、总胆固醇(TC)、脂蛋白(a)[LP(a)]、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)与总胆红素(TBIL)等指标,并结合临床诊断对结果进行分析。结果 40例anti-HCV阳性患者中,HCV-RNA阳性率为66.7%,hs-CRP、apoAl、apoB、TG、TC、LP(a)、HDL-C、LDL-C、ALT、AST和TBIL的含量与正常健康人差异有统计学意义(P0.05)。结论 anti-HVC和HCV-RNA的联合检测,对于丙型肝炎的诊断及治疗具有重要的实用价值及临床意义,如与生化等指标同时检测,将会对临床更有意义。     

丙型肝炎病毒总核心抗原测定方法及临床应用

目的建立丙型肝炎病毒总核心抗原（总HCV-cAg）酶联免疫测定方法,并对相关的临床样品进行测定。方法通过对样品进行裂解处理,采用酶联免疫试剂盒（ELISA）检测总HCV-cAg,对201名抗-HCV阳性者血清进行总HCV—cAg检测,同时进一步作HCV-RNA检测,其中176例采用荧光定量PCR（FQ—PCR）,25例采用逆转录PCR（RT—PCR）检测HCV．RNA。结果共检测201份血清标本,其中经PCR测定HCVRNA阳性88人份,阳性率43．8％;总HCV—cAg阳性71份,阳性率35．3％。经统计学分析,总HCV—cAg检测和HCVRNA检测的阳性率的差异无统计学意义。结论建立的总HCV—cAg酶联免疫测定方法适合临床应用,尤其适合在缺少RT-PCR或荧光定量PCR的中小医院使用。     

血清中HCV抗体与HCV-RNA检测在临床中的应用

目的探讨丙型肝炎病毒抗体与RNA在临床中的应用价值。方法用ELISA法检测934例丙型肝炎可疑患者病毒抗体,用实时荧光定量PCR检测379例丙型肝炎可疑患者病毒RNA。结果934例患者HCV抗体阳性率为19.16%,379例HCV-RNA阳性率为43.01%。53例为两者均做的病人,RNA和Ab均阳性的占52.8%(28/53),均阴性占15.1%(8/53),两者的一致率达67.9%,Ab的阳性率为81.1%(43/53),RNA的阳性率为56.6%(30/53)。结论HCV抗体联合HCV-RNA检测对丙型肝炎的临床诊断有一定的价值。     

A new assay for hepatitis C virus (HCV) core antigen detection: an alternative to nucleic acid technologies in positive or indeterminate anti-HCV subjects?

Markers of viral replication are fundamental tools for understanding the mode of transmission, diagnosis and management of hepatitis C virus (HCV) infection. A new enzyme-linked immunosorbent assay for quantitative detection of free and complexed HCV core antigen (HCV Ag) has been developed. The aim of this study is to evaluate the clinical performance of the new test and compare it with the most widely used commercially available RT-PCR-based assay. To determine the cut-off value we tested 60 samples from anti-HCV negative samples and selected a qualitative cut-off value of 3 pg/ml. To evaluate the usefulness of the new assay in confirming serologically indeterminate results we collected 62 sera. To evaluate the HCV Ag and HCV-RNA relationship we tested 245 samples from patients with different clinical conditions. The results of 61 out of 62 (98.4%) anti-HCV indeterminate samples were found to agree, whereas only one serum was found to be RT-PCR positive and HCV Ag negative. We also found the results to agree in 77.6% (190/245) of the samples from infected patients, while we observed higher agreement in untreated patients, both with and without evidence of liver damage. The correlation coefficient (r) observed between HCV Ag and HCV-RNA was 0.88. The regression line meets the cut-off value at an HCV-RNA concentration of approximately 40,000 IU/ml. In conclusion, we found that the results from the total HCV Ag test agree with the RT-PCR results and therefore we believe that this test could become a useful tool for the diagnosis and management of HCV infection.     

HCV and GBV-c/HGV infection in HIV positive patients in southern Italy

Flaviviridae–hepatitis C virus (HCV) and GB virus C/hepatitis G virus (GBV-C/HGV) – and human immunodeficiency virus (HIV) frequently show similar modes of transmission. HCV and GBV-C/HGV infection was assessed in 134 consecutive patients with evidence of HIV infection, living in Campania, Italy. Data obtained from this cohort were compared with those obtained from 252 age- and sex-matched HCV infected patients without evidence of HIV infection (HCV control group). Following enzymatic immunoassays, samples were tested for the presence of HCV-RNA by RT-PCR. The HCV-RNA positive sera were genotyped by LiPA procedure. The prevalence of HCV infection in HIV patients was 19.40% and the largest group of HIV–HCV co-infected patients (84.62%) was represented by intravenous drug users (IVDU). The distribution of HCV genotypes in HIV–HCV patients was different, compared to that observed in HCV control group. HCV genotypes 1a (50%) and 3a (23.08%) were more frequently detected in HIV–HCV patients, compared to HCV control group (5.16 and 5.56% for 1a and 3a, respectively). Conversely, HCV genotypes 1b (55.70%) and 2a/2c (30.26%) were more represented in HCV control group, compared to HIV–HCV patients (15.38 and 0% for 1b and 2a/2c, respectively). GBV-C/HGV seroprevalence was 41.04% in HIV patients and 6.54% in healthy control individuals. Differently from HCV, GBV-C/HGV infection did not correlate to a preferential risk behaviour in the HIV cohort. Comparative analysis of HCV and GBV-C/HGV infection indicates that the use of injecting drugs might play a key role in the epidemiology of HCV and, in particular, of 1a and 3a HCV genotypes, in HIV patients.     

同时检测4种传染病病原体的蛋白质微阵列研究

[目的]建立1种用蛋白质微阵列法可同时检测血清中艾滋病病毒（HIV）抗体、丙型肝炎病毒（HCV）抗体、梅毒螺旋体（TP）抗体和乙型肝炎病毒表面抗原的方法。[方法]将基因工程HIV、HCV、TP重组抗原和乙肝病毒单克隆抗体共价结合于固相载体玻片上，制成蛋白质微阵列。血清样本经稀释、加样、孵育、洗涤后，加上Cy3荧光标记物，用激光芯片扫描系统对蛋白质微阵列进行扫描成像。将获得的图像用Imagene专用分析软件进行分析，所获数据根据cutoff值自动生成判断结果。用此蛋白微阵列系统检测了100份4个项目皆阴性的血清标本，确定了其cutoff值。检测了经酶联免疫吸附试验（EUSA）筛选出4个项目皆阴性的标本40例；艾滋病病毒抗体阳性标本30例；乙肝表面抗原、丙肝抗体、梅毒特异性抗体阳性血清各100份。[结果]蛋白质微阵列法检测艾滋病病毒抗体、丙肝抗体和乙型肝炎表面抗原的阳性和阴性标本的符合率皆为100％，梅毒特异性抗体阳性符合率为97％，阴性符合率为100％。[结论]蛋白质微阵列法检测HIV、HCV、TP和HBsAg与EUSA法检测结果具有高度的符合率。该法具有高通量、快速、特异性强、操作简便等特点，适用于大规模样本的分析，在卫生检疫传染病检测方面具有应用和推广价值。     

丙型肝炎病毒抗体光密度值与丙型肝炎病毒核酸的关系

［目的］了解抗ＨＣＶ的ＯＤ值与ＨＣＶ－ＲＮＡ的关系。为丙型肝炎的诊断提供科学依据。［方法］从太原市中心血站、太原市卫生防疫站、山西省妇女劳动教养所、本站性病门诊、山西医科大学第一医院采集抗ＨＣＶ阳性血清２９６份，筛选出抗ＨＣＶ阳性的血清２２８份，用逆转录巢式多聚酶链反应（ＲＴ－nestPCR)进行ＨＣＶ－ＲＮＡ的检测。［结果］２２８份抗ＨＣＶ阳性标本中，检出ＨＣＶ－ＲＮＡ阳性者１６１例，阳性率为７０.6%，在抗ＨＣＶ的效价增高到一定值（ＯＤ／ＣＯ≥8.0)时，ＨＣＶ－ＲＮＡ检出率增高较明显，结果还显示不同人群抗ＨＣＶ阳性者的ＨＣＶ－ＲＮＡ检出率有差别，献血员、丙型肝炎患者的ＨＣＶ－ＲＮＡ检出率较高（８５.25%和８１82%）。［结论 ］当抗ＨＣＶ阳性的血清ＯＤ／ＣＯ≥8.0时，ＨＣＶ－ＲＮＡ很可能也为阳性。