丙肝抗体联合丙肝核心抗原在丙型肝炎诊断中的作用

目的 分析丙肝抗体与丙肝核心抗原联合诊断丙型肝炎的临床效果.方法 选取180例2017年1月—2018年6月在本单位采集的单试剂丙型肝炎阳性的献血者血样,对血清标本的丙肝抗体、丙肝核心抗原进行检测,分析不同方法 诊断丙型肝炎的效果.结果 以重组免疫印迹试验结果 为诊断标准,丙肝抗体诊断丙型肝炎的灵敏度、特异度、阳性预测...     

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.     

登革热抗原抗体联合检测与双抗体检测效能比较

目的 比较抗原抗体联合检测与双抗体快速检测2种方法检测登革热的效能,为登革热快速诊断提供依据。方法 收集登革热临床诊断病例血清样本449份(病例组)、流行病学和(或)临床上判断与登革热无关血清样本689份(阴性对照组),分别进行抗原抗体联合检测(IgM、IgG和NS1抗原)及双抗体检测(IgG和IgM),以临床诊断为金标准,对2种方法检测结果进行描述分析和一致性分析。结果 纳入研究的449例病例组和689例阴性对照组中,联合检测阳性率(34.1%)高于双抗检测(29.7%),且均低于临床诊断阳性率(39.4%),差异均有统计学意义(χ²值分别为20.61、7.03和51.33,P值均<0.01)。联合检测结果与实际临床诊断一致性(kappa=0.863)高于双抗检测(kappa=0.745),联合与双抗检测法结果一致性较好(kappa=0.729)。阴性对照组有9份样本经联合和(或)双抗检测法检测为登革热阳性,其中4例(3例发热待排查病例,1例体检人员)2种方法均为阳性,增补为登革热临床诊断病例。结论 抗原抗体联合胶体金检测登革热不仅方便、快捷、经济,与临床...     

389例丙肝病毒感染者合并乙肝病毒感染的调查

目的 通过对丙肝病毒感染者和正常体检者间的乙肝病毒5种标记物检出率的比较，探讨丙肝病毒感染与乙肝病毒感染的关系。方法 ELISA法检测乙肝病毒五项指标。结果 HBV5种标记物中，丙肝组与对照组间的抗－HBC检出率有统计学意义，且有相互关联，其列联系数为0．186。结论 丙肝感染群体的乙肝病毒核心抗体阳性率明显高于一般人群。     

丙型肝炎病毒核心抗原检测对缩短抗体检测窗口期作用的研究

目的探讨HCV Core Ag缩短Ab检测窗口期的作用。方法通过询问病史筛选存在丙肝危险因素人群35例,通过cAg、Ab检测结果研究缩短窗口期情况。结果 HCV cAg早期辅助临床诊断丙肝比Ab检测(2011年前)方法缩短约36.5天,Ab检测(2011年后)方法缩短约33.7天。结论 HCV Core Ag实验可有效缩短窗口期,为临床提供可信结果。     

三类人及时查丙肝抗体

来自福建福州市的高先生今年62岁,两个多月前,高先生出现乏力、无食欲、消瘦等症状,到医院检查发现得了丙肝,医生告诉他,丙肝病毒在他体内应该已存在较长时间,要尽快接受治疗,否则肝功能将进一步损坏。高先生感到非常不解,自己为何会突然查出有丙肝呢？     

丙型肝炎病毒核心抗原在丙型肝炎早期诊断中价值

目的 评估丙型肝炎病毒(HCV)核心抗原测定方法,用于丙型肝炎(以下简称丙肝)早期诊断的特异性、敏感性以及在缩短HCV窗口期诊断时间上的可行性及临床价值;为该方法用于临床提供科学实验依据.方法 特异性评估:对1 948份未感染HCV阴性血样(1 500份供血者的血样、400份健康检查的血样及48份非丙肝患者血样)进行HCV血清学及分子生物学检测;首先用2种酶联免疫吸附试验(ELISA)对HCV抗体进行测定,所有标本全部阴性;然后将上述标本全部进行HCV核心抗原测定,凡初次阳性的标本复查1次;并用HCV荧光定量聚合酶链反应(HCV荧光定量PCR)进行确证.敏感性及丙肝早期诊断应用价值评估:对55例丙肝患者及9套HCV血清panel进行HCV核心抗原测定;阴性结果复查,并用HCV荧光定量PCR确证.方法内及方法间精密度评估:对2份HCV RNA阴性、HCV抗体阴性的标本进行HCV核心抗原重复测定,并计算均数及CV值.结果 非HCV感染标本测定结果表明,1 500份血库标本中有8份在初次HCV核心抗原测定时出现阳性(0.53％),400份健康检查血样HCV核心抗原测定结果全阴,48例住院非丙肝患者HCV核心抗原检测结果只有1例阳性(2.08％).上述抗原阳性血样的HCV荧光定量PCR均为阴性,特异性为99.54％;96份HCV感染的血样(55例来自丙肝患者;41份来自9套HCV血清panel)进行HCV核心抗原测定及HCV荧光定量RNA测定,结果表明有92份HCV核心抗原测定阳性,敏感性为95.83％;对9套HCV血清panel测定结果表明,与HCV抗体方法相比,HCV抗原测定及HCV荧光定量PCR都能显著地将HCV感染窗口期平均缩短至36 d;方法内及方法间精密度的CV值均小于10％.结论 HCV核心抗原ELISA测定法具有与传统HCV荧光定量PCR相同的高度敏感性及特异性,可以明显缩短HCV早期感染窗口期,有效阻止HCV的传播.     

新型丙肝病毒分片段抗体检测试剂盒（蛋白芯片）问世

由上海裕隆生物科技有限公司研发的、被行内专家评为“国内首创、国际领先”的丙型肝炎病毒分片段抗体检测试剂盒(蛋白芯片)，日前获国家食品药品监督管理局颁发的新药证书。它标志着我国生物芯片技术向临床免疫检测领域的又一次成功突破。     

丙型肝炎病毒抗体阴性献血人群中丙型肝炎病毒核心抗原检测的研究

目的观察丙型肝炎病毒(HCV)抗体阴性献血人群HCV核心抗原的流行情况,初步评估经HCV抗体检测后输血传播HCV的潜在风险. 方法 抗HCV抗体和HCV核心抗原检测采用ELISA试剂盒,丙氨酸氨基转移酶检测采用速率法,HCV RNA检测采用实时荧光PCR检测试剂盒. 结果在抗HCV检测阴性的1 758份无偿献血者标本中,HCV核心抗原检测有3份阳性,阳性率为0.17%;HCV核心抗原阳性的3份样本HCV RNA检测为阴性. 结论结果提示抗HCV检测后,经血液传播HCV的潜在风险已经很低.     

Prevalence of antibodies to hepatitis C virus among family members of patients with chronic hepatitis C

In this study, 108 family members of 40 chronically HCV-infected patients (19 post-transfusion and 21 sporadic), and 45 families of 16 anti-HCV-negative index cases (control group) were tested for anti-HCV antibodies. Anti-HCV antibodies were found in 16 (14.8%) families of anti-HCV positive index cases (15% males and 14.6% females; p = NS), with no difference between families of index cases with post-transfusion and those with sporadic HCV infection. Out of the 16 anti-HCV positive family members, 12 (75%) had clinical and/or serological evidence of chronic liver damage. None of the control group subjects were anti-HCV-positive (p < 0.01). The rate of anti-HCV positivity was 34.4% among spouses, 14.3% among siblings, 16.7% among cohabitants and 2.3% among children; anti-HCV antibodies were not detected among parents. We found a positive correlation between the prevalence of anti-HCV antibodies among families and the severity of the HCV-related chronic liver damage of the index cases (p < 0.00005). In addition, to confirm that HCV infection and HCV-related chronic hepatitis may be transmitted intrafamiliarly, our findings also indicate that horizontal, especially sexual contact, is a more important route of HCV infection than vertical/perinatal transmission. Finally, the risk of acquiring HCV infection among families appears to be the highest when index cases are suffering from severe HCV-related chronic hepatitis.Corresponding author.     

我国部分地区丙型肝炎病毒基因分型研究

目的：研究中国丙型肝炎病毒（HCV）基因型分布。方法：应用AmplicorPCR试剂盒检测北京、青岛、固安和周口等地27例抗－HCV阳性的单采浆供血员和36例慢性丙型肝炎病人血清HCV RNA,并对其中36例HCV RNA阳性者用INNO－LiPA^TMHCV II试剂进行HCV基因分型。结果：抗－HCV阳性的单采浆供血员和慢性丙型肝炎病人HCV RNA检出率0分别为48.15％（13／27）和69.44%(25／36）,平均为60.32％（38／63）。对13例HCV RNA阳性的单采浆供血员和23例慢性丙型肝炎病人HCV基因分型结果表明,29例（80.55％）为1b型,5例（13.89％）为2型,另2例（5.56％）不能被分型。结论：上述调查地区流行的HCV基因型以1b型占优势。     

献血感染丙型肝炎人群转归分析

目的研究献血感染丙型肝炎病毒（HCV）人群16年后的转归。方法对该人群问卷收集一般情况,肝脏B超检查;采集5ml静脉血,进行丙型肝炎病毒抗体、RNA、谷丙转氨酶（ALT）和血清生化指标（透明质酸、Ⅲ型前胶原肽、Ⅳ型胶原）检测。结果162名研究对象尚未出现晚期肝病患者。抗-HCV阳性率95.68%,RNA阳性率77.78%,ALT异常率20.37%。不同急性期症状感染者,16年后抗-HCV阳性率、ALT异常率和RNA阳性率差异无统计学意义。经性别分层分析,女性感染年龄〉40岁抗体较易阴转（χMH^2=8.26,P=0.04）。结论HCV感染16年后,不同急性期症状感染者转归无差异,病毒清除与性别和初始感染年龄有关。     

宜兴地区丙型肝炎病毒基因型及变异情况分析

目的了解江苏宜兴地区丙型肝炎病毒（HCV）基因型分布特点和变异情况对疾病的影响。方法对宜兴市人民医院收治的166例HCV抗体阳性患者血清进行HCVRNA检测，采用5′非编码区（5′NCR）1，2，3，1b型特异性引物。对阳性标本进行Simmonds分型。选取未经干扰素治疗的43份慢性丙型肝炎患者的HCVRNA阳性血清，采用RT—PCR扩增HCV包膜区（E2），单链构象多态性聚合酶链反应法（SSCP）获得HCV准种条带数，准种数〉2为复杂准种，比较不同肝病活动度患者的准种复杂情况。结果166份血清中，有102份为HCVRNA阳性。其中1b型86份占84．3％，2型6份占5．9％，1b／2型的混合感染5份占4．9％，不能分型的5份占4．9％。男性和女性在基因型总体分布差异有统计学意义。在单一型感染和混合型感染的分布差异亦有统计学意义，而在不同年龄、不同临床类型的肝炎中分布差异无统计学意义。HCV包膜区准种复杂与否和丙氨酸转氨酶（ALT）的正常与否具有相关性。结论江苏宜兴地区的HCV感染主要为1b型，病毒的变异程度与ALT成正相关。     

Inhibition of hepatitis C virus core and NS3 protein expression by siRNA

Objective To construct two kinds of plasmids with one for gene expression and the other for RNA expression. Methods Hepatitis C virus (HCV) core and NS3 genes were firstly inserted into the upstream of the reporter genes marked with Rellina (hRluc) luciferase and Firefly luciferase (hluc + ). And then the established Plasmids were cotransfected into HEK293 cells. Six short hairpin RNAs targeted HCV core and NS3 genes were designed, and expressed by siRNA expression vector. Inhibition effects with the luminescent intensity were observed. Results Light density in the cells cotransfected with core gene and anti-Core siRNA was obviously weaker, meanwhile, those with NS3 gene and anti-NS3 siRNA had no difference in contrast with the control group. Transfected the anti-NS3 siRNA into the stable expression cells and detected them by real-time PCR and western-blot. The four siRNAs have efficiently suppressed the expression of NS3 at some level. Conclusion The results demonstrated that siRNA was effective in inhibiting HCV protein expression, and may have therapeutic potential to limit HCV replication in chronically infected patients.     

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。     

Neutralizing antibodies and broad,functional T cell immune response following immunization with hepatitis C virus proteins-based vaccine formulation

HCV is a worldwide health problem despite the recent advances in the development of more effective therapies. No preventive vaccine is available against this pathogen. However, non-sterilizing immunity has been demonstrated and supports the potential success of HCV vaccines. Induction of cross-neutralizing antibodies and T cell responses targeting several conserved epitopes, have been related to hepatitis C virus (HCV) clearance. Therefore, in this work, the immunogenicity of a preparation (MixprotHC) based on protein variants of HCV Core, E1, E2 and NS3 was evaluated in mice and monkeys. IgG from MixprotHC immunized mice and monkeys neutralized the infectivity of heterologous HCVcc. Moreover, strong CD4+ and CD8+ T cells proliferative and IFN-γ secretion responses were elicited against HCV proteins. Remarkably, immunization with MixprotHC induced control of viremia in a surrogate challenge model in mice. These results suggest that MixprotHC might constitute an effective immunogen against HCV in humans with potential for reducing the likelihood of immune escape and viral persistence.     

Characterization of the humoral and cellular immune responses against hepatitis C virus core induced by DNA-based immunization

Hepatitis C Virus (HCV) causes most cases of posttransfusion hepatitis. Chronic HCV infection is highly related to chronic hepatitis, cirrhosis and hepatocellular carcinoma. Current therapies are only minimally effective and no vaccine has been developed. DNA-based immunization could be of prophylactic and therapeutic value for HCV infection. By intramuscular inoculation in BALB/c mice with an HCV recombinant plasmid pCI-HCV-C, we found significant levels of IgM antibody, but no significant IgG rise. After boost the immunized mice with recombinant HCV-core protein (cp1-10; 1-164aa), the anticore IgG, verified by Western-blotting, rose rapidly, which was two weeks earlier than that with control plasmid. Spleen cells from pCI-HCV-C immunized mice gave higher proliferation index (PI) than control (P < 0.05). The PI of cp1-10 boosted mice was even higher. Proliferation blocking assay with mAb proved the responding cell to be of CD4+ CD8- phenotype, supporting specific priming of T helper cells. A 51Cr-releasing CTL assay specific for HCV-core was developed, and a specific CTL response against HCV-core was demonstrated in both pCI-HCV-C immunized mice and mice boosted with cp1-10. Strong cytotoxic activity against peptide-pulsed p815 cells (H-2d), but not EL-4 cells (H-2b), suggested MHC class I restriction of the CTL activity. Blocking of CTL with mAb proved the effector cells to be of CD4- CD8+. Three CTL epitopes in HCV-core protein were demonstrated. We failed to detect CTL when immunized only with core protein. The results suggested that vaccination with HCV-core derived DNA sequences could be an effective method to induce humoral and cellular immune responses to HCV.     

Development of antibody to hepatitis C virus (HCV) in acute and chronic non-A,non-B post-transfusion hepatitis

The antibody to hepatitis C virus (anti-HCV) was measured by an immunoassay in 507 serum samples from 94 patients with acute and chronic post-transfusion non-A, non-B hepatitis (NANB) and in 436 healthy blood donors. Anti-HCV was found in 70.8 of patients with acute hepatitis, in 78.2 with chronic hepatitis, and in 1.4 of healthy blood donors. In acute hepatitis, anti-HCV appeared in the serum from 4 to 34 weeks after transfusion and from 1 to 30 weeks after the onset of the overt disease. Three patients with resolving hepatitis (21%) and 2 who developed chronic hepatitis (10%) lost anti-HCV during a mean follow-up period of 28 months. Among the 36 patients with chronic hepatitis, 2 (6%) lost anti-HCV after 12 months and 8 years respectively. These data indicate that in recent years HCV has been the major etiologic agent of acute and chronic transfusion-associated hepatitis (TAH) in our geographical area. The late appearance of anti-HCV from the onset of clinical and biochemical signs of acute hepatitis in more than 70% of patients limits the diagnostic utility of this assay for an earlier serological diagnosis of acute NANB hepatitis. Additional studies are required to determine the diagnostic significance of this antibody in chronic NANB hepatitis.Corresponding author.     

丙型肝炎病毒体外复制及其动物模型

本文简要介绍了丙型肝炎病毒在肝细胞系和淋巴细胞系中的复制情况,着重介绍了丙型肝炎病毒复制子在HuH-7细胞中的复制规律;阐述了黑猩猩、转基因小鼠和嵌合肝脏小鼠作为丙型肝炎病毒研究的动物模型所存在的优缺点,以及树鼩作为丙型肝炎病毒研究的动物模型的可能性。