新型丙型肝炎病毒抗体检测试剂Elecsys anti-HCV Ⅱ的性能评价与比较

目的评价新上市的丙型肝炎病毒抗体(抗-HCV)筛查试剂Elecsys anti-HCV Ⅱ的性能,并与其他两种临床上广泛应用的同类试剂进行性能比较。 方法使用罗氏Elecsys anti-HCV Ⅱ、雅培Architect anti-HCV和强生Vitros anti-HCV 3种试剂,平行检测4个HCV血清转换盘、861份临床常规样本、100份抗-HCV检测临界阳性样本及178份HIV感染患者样本。抗-HCV确诊试验为重组免疫印迹法(RIBA 3.0)或HCV RNA定量检测。此外,使用Elecsys anti-HCV Ⅱ检测203份不同HCV基因型样本以评价其基因型覆盖度。 结果相比Architect和Vitros anti-HCV,Elecsys anti-HCV II可提前7～14d检测到HCV感染后抗-HCV的产生;在检测临床常规样本(包括抗-HCV临界阳性样本)时,Elecsys anti-HCV Ⅱ有100%的敏感度及良好的特异度;70.22%(125/178)HIV感染患者样本为HCV RNA阳性,Elecsys anti-HCV Ⅱ可检测出其中97.60%(122/125)样本中的抗-HCV;Elecsys anti-HCV Ⅱ可能低估3b型样本的抗-HCV水平。 结论Elecsys anti-HCV Ⅱ可进一步缩短HCV感染后的检测窗口期,可灵敏、特异地检测临床样本包括免疫缺陷患者样本中的抗-HCV,适用于临床HCV感染的筛查,但对其检测特定HCV基因型样本的性能尚需纳入更多样本深入研究。     

HCV分片段抗原EIA与重组免疫印迹法分析检测HCV抗体的比较研究

【目的】探讨国产丙型肝炎病毒(HCV)分片段抗体试剂的敏感性和特异性,为丙型肝炎的早期诊断和替代昂贵的进口重组免疫印迹法确认试剂(HCV RIBA3.0)提供实验依据。【方法】用HCV分片段抗体试剂检测经确认的标本中HCV-C、NS3、NS4、NS5抗体并与HCV RIBA3.0比较。【结果】82份样品中80份判定为阳性,阳性率为97.56%,2份判定为可疑,为2.4%,40份阴性样品中有2份判定可疑,为5.00%。【结论】国产分片段抗体检测试剂检测HCV抗体,与美国第三代RIBA3.0试剂的检测符合率基本一致,有低价高效的临床实用价值。     

3组国产ELISA抗-HCV初复检试剂的检测效果评价

丙型肝炎病毒（hepatitis C virus，HCV）是我国输血后肝炎（post—transfusion hepatitis，PTH）的主要病因，PTH中90％是丙型肝炎患者，HCV感染常导致肝硬化，其中部分患者可发展为肝细胞癌。选择灵敏度、特异性高、质量稳定的初复检试剂进行血液检测，是有效控制HCV经输血传播的重要手段。为了有效避免抗-HCV阳性血液标本漏检，     

献血者抗-HCV ELISA阳性结果RIBA分析

笔者将本中心对献血者丙型肝炎抗体筛选所采用两种试剂同时阳性或仅一种试剂阳性的标本 ,采用RIBA法进行抗原区带分析 ,现将 1年的分析结果报告如下。1　材料与方法1 1　材料　 2 0 0 1年 5月～ 2 0 0 2年 5月献血者标本 4 4 0 0 7份 ,每份标本均采用ORTHO及华美抗 HCV试剂同时作抗 HCV筛选 ,收集两种试剂检测同时阳性或仅一种试剂阳性的标本 12 2份 ;CHIRONRIBAHCV 3.0SIA试纸条。1.2　方法　按照CHIRONRIBAHCV 3.0SIA说明书操作 ,在CHIRONRIBA分析仪上作抗体原区代分析。2　结果试验的内控带IgG条带Ⅱ颜色明显强…     

丙型肝炎病毒抗体诊断试剂盒(EIAgen HCV Ab Kit)的可信度分析

目的 分析丙型肝炎病毒(HCV)抗体酶联免疫吸附试验(ELISA)诊断试剂盒(EIAgen HCV Ab Kit)的可信度.方法 应用考核试剂EIAgen HCV Ab Kit和参比试剂Ortho HCV 3.0 ELISA对2 881例样本进行检测,检测结果不一致时,应用重组免疫印迹试验(RIBA)确证.所用试剂为RIBA(R)HCV 3.0 SIA或核酸试验(NAT),并对结果进行综合分析.结果 2 881例样本中,阳性样本333例,阴性样本2 539例,9例不确定的样本被剔除.考核试剂检测真阳性333例,无假阴性结果,真阴性2 527例,假阳性12例.考核试剂灵敏度为100.00%,高于参比试剂;特异性为99.53%,略低于参比试剂.考核试剂对部分国内常见HCV基因型的灵敏度为100.00%,对于非HCV感染的病毒性肝炎、自身免疫性疾病及妊娠样本具有良好的特异性,特异性均为100.00%.溶血、脂血样本对考核试剂检测结果无影响.考核试剂阳性预测值为96.52%,阴性预测值为100.00%,符合率为99.58%.考核试剂S/CO≥5.9的样本301例,其中用参比试剂检测S/CO≥3.8占88.70%;考核试剂S/CO＜5.9的样本32例,其中用参比试剂检测S/CO＜3.8占87.50%.结论 试剂EIAgen HCV Ab Kit灵敏度100.00%,特异性较好,是一种优质的抗-HCVELISA试剂,尤其适用于阳性样本的筛选.采用EIAgen HCV Ab Kit试剂检测样本,当S/CO≥5.9时,样本的阳性预测值比较高.     

肝脂肪变性促进慢性丙型肝炎病程进展的研究

目的 研究脂肪变性在慢性丙型肝炎疾病进程中的作用.方法 收集治疗前行肝穿刺病理检查的慢性丙型肝炎患者159例,按照感染HCV基因亚型分为基因1b型组、基因2a型组和其他基因型组,荧光定量PCR法检测所有病例血清HCV载量,组织学评估各组穿刺肝组织炎症坏死、纤维化和肝细胞脂肪变性程度.结果 HCV基因1b型和2a型感染患者占总病例的65.41%,63.52%(101/159)的慢性丙型肝炎患者发生肝细胞脂肪变性,不同基因型组间脂肪变性程度无统计学差异.不同基因型组HCV慢性感染者间炎症活动度、纤维化程度和脂肪变性程度无统计学差异(P均>0.05).肝脂肪变性与肝纤维化和炎症活动度均密切相关(r值分别为0.34和0.29,P均<0.01),但HCV感染病毒量与脂肪变性、肝纤维化和炎症活动度间无明显相关性.结论 脂肪变性促进慢性丙型肝炎病程进展.     

臭氧自血回输治疗丙型肝炎患者实验室检测分析及临床疗效评价

目的 通过观察臭氧自血回输疗法治疗丙型病毒性肝炎患者的实验室检测分析临床疗效及安全性,以分析其临床应用价值.方法 选择丙型肝炎患者133例,随机分为治疗组63例、对照组70例;两组患者均给予干扰素α-2b（Interferonα-2b）500万国际单位肌注、每周3次,并联合利巴韦林片300 mg、每日3次口服;治疗组患者加用臭氧自血回输疗法、每周3次治疗.治疗12周后,分别观察、比较其血清丙型肝炎病毒核酸（Hepatitis C virus ribonucleic acid,HCV RNA）水平的变化、血清病毒学应答率、生化学应答率及不良反应发生率.结果 治疗12周,两组患者血清HCV RNA水平中位数均较治疗前明显下降;治疗组血清HCV RNA水平下降幅度大于对照组,血清生化学指标优于对照组,差异均有统计学意义（P＜0.05）;血清病毒学应答率及生化学应答率,两组间差异无统计学意义（D0.05）.两组均未发生不良事件.结论 实验室检测结果分析得出臭氧自血回输疗法治疗丙型肝炎,可以使患者血清HCV RNA水平明显下降,并能促进患者肝功能恢复,安全性好;可用于丙型肝炎的辅助治疗.     

不同类型肝病患者血清抗-HCV检测结果分析

目的探讨各种肝病患者血清丙型肝炎病毒抗体(抗-HCV)阳性率以及感染状况,研究其临床价值。方法采用酶联免疫吸附法检测1 024例各种肝病患者血清抗-HCV。结果 1 024例肝病患者抗-HCV阳性率为33.59%(344/1024),阳性率由高到低依次为丙型肝炎(65.22%)、乙丙型肝炎(48.89%)、肝癌(35.71%)、肝硬化(32.37%)、慢性乙肝(28.66%)、黄疸型肝炎(19.26%)、重型肝炎(16.67%)、甲型肝炎(8.82%);男性和女性肝病患者抗-HCV阳性率分别为24.86%(176/708)、53.16%(168/316),阳性率差异有统计学意义(χ2=78.47,P<0.05)。结论各种肝病患者均检出抗-HCV,以丙型肝炎、乙型肝炎、肝癌及肝硬化患者阳性率较高;女性肝病患者HCV感染率高于男性。     

白细胞介素17在慢性丙型肝炎患者免疫损伤中的作用

目的 探讨白细胞介素17（IL-17）在慢性丙型肝炎患者免疫损伤中的作用.方法 选择22例慢性丙型肝炎患者作为HCV组,20例健康志愿者作为对照组.分别采用实时聚合酶链反应（RT-PCR）、酶联免疫吸附测定（ELISA）检测血清HCV-RNA及IL-17,其余检测指标包括血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、总胆红素（TBIL）、凝血酶原时间（PT）、肌酐.结果 HCV组患者血清IL-17水平明显高于对照组（P＜0.05）.HCV组患者血清ALT、AST及HCV-RNA与血清IL-17水平显著相关（P＜0.05）.结论 IL-17可能参与了慢性丙型肝炎的免疫学发病机制.     

某地区丙型病毒性肝炎患者HCV基因分型研究

目的 了解该地区丙型病毒性肝炎(以下简称丙型肝炎)患者丙型肝炎病毒(HCV)基因分型情况.方法 分别以ELISA和重组免疫印迹试进行血清标本抗HCV抗体筛查和确认,对抗HCV抗体阳性标本用实时荧光定量PCR进行病毒载量检测.对病毒载量大于103 copy/mL的标本用多重PCR进行HCV基因分型.结果 125例抗HCV抗体阳性标本中,HCV基因型主要为1b、2a、3a、1c、2c的检出率分别为58.4%、21.6%、3.2%、4.0%和8.0%.结论 该地区HCV感染主要以1b型为主,其次是2a和3a型,检出其他地区少见的1c和2c型,说明该地区丙型肝炎流行的基因型呈多样性.     

Detection of hepatitis C virus infection with recombinant immunoblot assay,synthetic immunoblot assay,and polymerase chain reaction

The newly developed immunoblot assay, RIBA SIA (recombinant and synthetic polypeptide immunoblot assay), Chiron, Calif., was compared with the commercially available second generation recombinant immunoblot assay (RIBA-2) for the detection of antibody to hepatitis C virus (anti-HCV). The two immunoblot tests were also compared with the polymerase chain reaction (PCR) for the detection of HCV RNA. Ninety-one percent of samples reactive by RIBA-2 were positive for anti-HCV by RIBA SIA. A total of 31% of RIBA-2 indeterminate samples became reactive by RIBA SIA, 24% became non-reactive, and 45% remained the same. Samples reactive by RIBA-2 or SIA from different risk groups, were mostly positive (67-100%) by PCR for HCV RNA. All indeterminate samples from hemophiliacs and intravenous drug users were PCR positive. RIBA SIA is more sensitive and specific than RIBA-2 and correlates well with PCR results © 1993 Wiley-Liss, Inc.     

ORTHO抗-HCV酶联免疫试剂2种孵育试验程序在血液筛查中的比较研究

目的确定1种3代抗-HCV酶免试剂(ORTHO HCV 3.0 ELISA试剂)2种孵育试验过程在血液筛查中是否存在差异。方法随机留取常规献血者血液筛查中检出的抗-HCV反应性的血液标本180(人)份作HCVRNA检测,并用RIBA和不同于血站常规抗-HCV筛查的酶免试剂作抗-HCV复测;依据ORTHO HCV 3.0 ELISA检测试剂说明书中的长、短2种孵育检测程序,同时作对比检测。结果 180份初筛抗-HCV阳性标本中,有16份ORTHO HCV 3.0 ELISA 2种检测程序的检测结果不一致,其中5份为短孵育试验反应性、11份为长孵育试验反应性,短孵育试验漏检2份被确证抗-HCV阳性标本。ORTHO HCV 3.0 ELISA试剂的长孵育检测程序灵敏度高于短孵育试验程序,2种试验程序的S/CO值分布没有差别,但RIBA不确定标本其S/CO值分布在一定的灰区范围内。结论在献血人群血液筛查中,采用具有更高灵敏度的长孵育酶免程序和设定合理的结果判定灰区,有助于预防输血传播HCV,提高血液的安全。     

HCV胶体金与ELISA法在献血者体检中的应用比较

目的应用丙型肝炎病毒（HCV）胶体金法检测无偿献血者,并与抗-HCV ELISA法比较。方法对2008年1月～2009年12月的5830例无偿献血者,先应用HCV胶体金试纸条检测,再分别用试剂1、试剂2两种抗-HCV ELISA试剂进行常规初检、复检。对HCV胶体金法阳性的标本、试剂1初检、试剂2复检阳性的标本,再进行HCV RNA RT-PCR荧光定量检测。结果在被检测的5830份标本中,HCV胶体金法阳性有11份,抗-HCV ELISA试剂1初检阳性12份、试剂2复检阳性13份;其中初检、复检均阳性的11份,仅初检阳性1份和仅复检阳性2份,HCV RNA RT-PCR荧光定量检测阳性11份。结论本组检验HCV胶体金法与HCV RNA RT-PCR荧光定量法结果符合率为100.0%,HCV胶体金试纸法假阳性率低,操作简便、快速,适合无偿献血者抗-HCV筛选。     

Lack of correlation between sensitivity characteristics of the tests for hepatitis C virus antibodies estimated with serially diluted and natural low-reactive control specimens

Sensitivity characteristics of seven commercial ELISA test systems for the detection of antibodies to hepatitis C virus were assessed using control panels consisting of: (i) serial dilutions of pooled sera highly reactive for anti-HCV; (ii) serial dilutions of RIBA 3.0 HCV SIA positive control; and (iii) natural (non-diluted, non-spiked) sera low-reactive for anti-HCV. "Dilutional sensitivity" values estimated with these two kinds of highly reactive samples did not coincide and were not found to correlate with the proportion of natural low-reactive specimens detected by each test. Thus, laboratories assessing sensitivity of anti-HCV ELISAs should take into consideration the nature and properties of the control material used. Natural low-reactive control specimens are preferable because they adequately reflect the real serological picture of early stage of HCV infection.     

Application of a new enzyme-linked immunosorbent assay for detection of total hepatitis C virus core antigen in blood donors

Recent studies have shown that total hepatitis C virus (HCV) core antigen, both free and antibody bound, is an accurate indirect marker of viral replication that can be used in clinical practice. The aim of the present study was to evaluate the performance of a new total HCV core antigen enzyme-linked immunosorbent assay (ELISA) for detection and quantification of total core antigen in blood donors, testing positive for anti-HCV antibodies and for prospective low-risk population screening. A population comprising 257 samples, from blood donors detected reactive for anti-HCV antibodies [137 recombinant immunoblot assay (RIBA) positive and 120 RIBA indeterminate], were tested by using a new total HCV core antigen ELISA. HCV-RNA was quantified by using quantitative polymerase chain reaction (PCR) assays in all RIBA-positive samples and RIBA-indeterminate samples that were positive for the total core antigen. Specificity of the assay was studied in 1070 healthy blood donors negative for anti-HCV antibodies. Compared with quantitative PCR assays, the total HCV core antigen assay showed 97.37% sensitivity. The three HCV-RNA-positive samples, which tested negative for the total core antigen, had a low viral load (< 1.4 x 10(4) IU mL(-1)). All samples with more than 1.4 x 10(4) IU mL(-1) of viral RNA were positive for total core antigen, independent of the HCV genotype. Concentration of total core antigen correlated significantly with those of HCV-RNA (r = 0.614, P < 0.0001). Overall specificity in freshly collected blood donor specimens was 99.63%. Our data indicate that the total HCV core antigen ELISA has a sensitivity close to PCR assays in diagnosing HCV infection in blood donors with anti-HCV antibodies and shows an excellent specificity in volunteer donors. This assay, in combination with anti-HCV antibodies screening tests, could be an alternative to molecular assays for HCV infection screening in blood donors.     

Anti-hepatitis C virus (anti-HCV) seroconversion in patients undergoing hemodialysis: comparison of second- and third-generation anti-HCV assays

BACKGROUND: The results obtained in sequential specimens from recently infected subjects generally provide the best means of comparing the sensitivity of assays. STUDY DESIGN AND METHODS: The sensitivity of second- and third-generation assays for antibody to hepatitis C virus (HCV) was compared on sequential specimens, generally collected at monthly intervals from 45 patients undergoing hemodialysis who seroconverted for HCV between 1980 and 1990. RESULTS: Fifteen patients (33%) were positive earlier in the third-generation enzyme-linked immunosorbent assay (ELISA), with a mean difference of 17 days (range, 7–30) between the last negative and the first positive specimens. At the first rise in alanine aminotransferase, and at its peak, 63 and 91 percent of the patients, respectively, were anti-HCV positive in the third-generation ELISA. Third-generation recombinant immunoblot assay (RIBA) reacted at the same time as third-generation ELISA. Of the first specimens that were positive in second-generation ELISA, 44 percent reacted and 56 percent were indeterminate in third-generation RIBA, while 10 percent reacted, 72 percent were indeterminate, and 18 percent did not react in second-generation RIBA. From the beginning to the end of the follow-up, antibody to c33c was the most prevalent, followed in descending order by antibody to c22-3, antibody to c100-3, and antibody to NS5: 56, 54, 26, and 18 percent, respectively, at time 0, and 100, 86, 83, and 31 percent, respectively, 12 months later. CONCLUSION: Third-generation assays (both ELISA and RIBA) were more sensitive than second-generation assays in the diagnosis of HCV infection, in that positive results were obtained earlier and a higher proportion of specimens were confirmed positive in RIBA testing.     

HCV抗体检测临界值附近样本传播HCV风险的研究

目的用尽可能低的消耗，达到HCV感染早期诊断，尽量缩短ELISA HCV抗体检测的“窗口期”，防止和减少经输血传播丙型肝炎的风险。方法采集初次检测抗-HCV（S／CO）样本测定值／临界值0．40～0．99的样本310份，采用“HCV抗体分片段酶联免疫确认试剂盒”进行补充旁证检测，对检测阳性及可疑阳性样本再进行HCV—PCR和RIBA及HCV—cAg检测。结果310份样本共检出单片段阳性样本25份，2个片段以上阳性样本3份，共计28份。HCV—PCR阳性3份，HCV—cAg阳性4份，RIBA检测3个条带阳性2份，1个条带阳性22份。28份样本中，抗-HCV分片段、PCR、HCV—cAg以及RIBA共同阳性1份，HCV—cAg与PCR共同阳性2份，HCV—cAg和RIBA共同1份。结论加强对抗-HCV检测阴性高值样本的重视，尽量缩短ELISA—HCV抗体检测“窗口期”。     

血友病A患者血制品治疗后HBV和HCV感染指标分析

目的对血友病A患者替代治疗后血液传播乙型、丙型肝炎病毒(HBV、HCV)的感染指标进行检测。方法对经本院确诊的35例血友病A患者采用酶联免疫吸附法(ELISA)检测抗-HCV、HBV六项指标。结果 35例血友病A患者的抗-HCV阳性率为88.6%,输血次数和输注血液制品为主的种类与患者抗-HCV阳性率有相关性(P<0.01)。HBV六项指标检查,其中5例患者抗-HBe阳性(占14.3%),明显低于抗-HCV阳性率。结论血友病A患者替代治疗输血次数越多,感染风险越大,而较少输注以冷沉淀为主的血液制品的患者,其感染风险相对较小,且目前HCV感染率明显高于HBV感染率。     

ELISA试剂检测抗HCV反应性结果分析

目的探讨丙型肝炎病毒抗体(抗HCV)进口酶联免疫吸附试验(ELISA)试剂在血液筛查中的效果及与国产试剂、重组免疫印迹试验(RIBA)确证检测的相关性。方法用RIBA确证试剂及国内销量前2名的国产抗HCV ELISA试剂对56例进口抗HCV ELISA试剂检测呈反应性的标本进行检测分析。结果 56例标本经RIBA确证结果为阳性12例,不确定18例,阴性26例。3种试剂均为反应性的9例标本中,确证阳性7例,不确定1例,阴性1例。结论目前市售的丙型肝炎ELISA试剂均存在较大的生物学假阳性,不同试剂间检测结果仍存在一定差异。为确保输血安全,建议有条件的采供血机构实验室抗HCV检测时以国产和进口试剂联合检测为佳。