丙型肝炎病毒检测方法的应用评价

目的评价酶联免疫吸附法（ELISA法）检测丙型肝炎病毒核心抗原（HCV—cAg）和抗体（HCV—Ab）以及反转录多聚酶链反应（RT—PCR）检测丙肝RNA,了解三种方法对丙肝病毒检测的优点和缺点。方法采用HCV—cAgELISA试剂盒．HCV—AbELISA试剂盒和丙肝病毒PCR检测试剂盒对来自临床的225例样本进行HCV—cAg、HCV—Ab和HCV—RNA检测。结果225例样本中检出抗HCV阳性28例,其中HCV—cAg阳性14例,RT—PCR阳性16例,剩余的197例抗HCV阴性标本中,检出HCV—cAg阳性3例,其中RT—PCR阳性2例。结论RT—PCR技术检测HCV—RNA是判断丙肝感染的最有效方法。HCV核心抗原检出时间早于抗体,联合运用抗HCV和HCV—cAg或者抗HCV和HCV—RNA检测HCV,能有效降低HCV—Ab检测带来的漏检风险。     

丙型肝炎病毒核心抗原检测的临床应用价值

目的探讨丙型肝炎病毒核心抗原（HCV—cAg）酶联免疫吸附试验（ELISA）检测方法的临床应用价值。方法对20例HCV抗体（HCV—Ab）阳性标本和200例HCV—Ab阴性标本进行HCV—cAg测定,同时采用核酸扩增进行HCV—RNA对照测定。结果20例HCV—Ab检测阳性的标本中HCV—cAg阳性8例,200例HCV—Ab检测阴性的标本中HCV．cAg阳性3例;以HCV—RNA聚合酶链反应（PCR）检测为对照,HCV—cAg检测试剂盒的敏感性为88．3％,特异性为99．5％。结论HCV—cAgELISA检测方法敏感性和特异性较好,HCV—Ab和HCV—cAg联合检测可起到互补作用,提高阳性检出率,有利于早期诊断。     

血清转换样本丙型肝炎病毒核酸、核心抗原及抗体检测对比研究

目的探讨丙型肝炎病毒(HCV)核心抗原酶联免疫诊断试剂检测早期HCV感染“窗口期”样本的可行性。方法用新研制的丙型肝炎病毒核心抗原(HCV-cAg)酶联免疫诊断试剂检测11份系列血浆转换样本的抗-HCV、HCV-cAg、HCV RNA,并与病毒核酸及抗体检测方法做对比研究。结果HCV-cAg较抗-HCV检出提前14~51d,平均32.7d;HCV RNA较抗-HCV检出提前19~51d,平均36.0d。结论HCV-cAg酶联免疫诊断试剂可用于HCV早期感染“窗口期”样本的检测,降低输血风险度。     

ELISA检测丙型肝炎病毒核心抗原的临床应用价值

【目的】探讨丙型肝炎病毒（HCV）核心抗原酶联免疫吸附试验（ELISA）检测方法的临床应用价值。【方法】采用HCV核心抗原ELISA试剂盒,对32例HCVRNA基因扩增试验检测为阳性的临床丙肝感染者血清进行检测,同时用抗-HCVELISA检测试剂盒作为对照进行比较。【结果】与HCVRNA PCR检测为对照,HCV抗原检测试剂盒的灵敏度为96．9％,特异性为96．7％。32例HCVRNAPCR检测阳性血清,用抗-HCV检测法进行检测,其中有5例阴性,将这5例阴性标本用HCV核心抗原检测法进行检测,其中4例为阳性。【结论】丙型肝炎核心抗原ELISA检测方法能缩短窗口期、敏感性高、特异性好、费用低廉,在临床上具有很好的推广前景。     

献血者丙型肝炎病毒（HCV）核心抗原的检测

目的：探讨在无偿献血者中开展丙型肝炎病毒核心抗原（HCV—CAg）检测的应用价值。方法：采用美国强生公司HCV核心抗原（HCV—CAg）酶联免疫分析（ELISA）试荆，对无偿献血者样进行了检测与结果分析。阳性者用荧光定量PCR方法进行HCV检测，并将抗体检测、抗原检测、病毒核酸检测（nucleic aeldtesting，NAT）的检测结果进行比较。结果：676份合格血样的HCV—CAg结果均为阴性。63份抗-HCV阳性中有11例HCV—CAg阳性，138例ALT阳性中有2例阳性，合计检出13例HCV C抗原阳性标本。经荧光定量PCR检测有12例HCVRNA阳性，其中11份抗-HCV阳性，ALT阳性的1份。结论：现行的抗-HCV检测方法有可能出现HCV感染的漏检，HCV—CAg检测方法可提高检测的灵敏度。应在献血者中开展该检验项目．     

丙型肝炎病毒核心抗原检测试剂的研制及初步应用

目的　研制丙型肝炎病毒核心抗原 (HCV cAg)ELISA检测试剂 ,用于诊断早期丙型肝炎。方法　用基因工程表达的HCV cAg ,免疫Balb/c小鼠 ,制备抗HCV cAg单克隆抗体 ,建立检测HCV cAg双抗体夹心酶联免疫吸附试验 (ELISA) ;以此检测 12 5份抗 HCV、抗 HIV、抗 TP阴性 ,但单项ALT高的献血者血浆。结果　 12 5份单项ALT高的献血者血浆中检出 9份HCV cAg阳性。结论　HCV cAg双抗体夹心ELISA试剂 ,有望用于早期丙型肝炎诊断。     

HCV核心抗原检测在血液筛查中的初步研究

目的 探讨丙型肝炎病毒核心抗原(HCV cAg)醇免检测试剂用于血液筛查的应用价值.方法 同时用抗HCV抗体和HCV cAg酶免检测试剂平行筛查血液,对HCV抗体阴性样本用Chiron Procleixa TMA系统检测HCV RNA.结果 1 762份无偿献血者样本中,抗HCV阳性12份(0.68％),HCV cAg阳性10份(0.57％),阳性检出率差异无统计学显著性意义(P＞0.05),抗HCV及HCV cAg同时阳性4份(0.23％).1 750份抗HCV阴性样本中,其中HCV cAg阳性样本6份,HCV RNA全为阴性.结论 在血液筛查中增加HCV cAg检测,对降低HCV的输血传播危险具有一定价值.     

丙型肝炎病毒核心抗原检测试剂的临床评价

[目的]对研制的丙型肝炎病毒核心抗原（HCV—cAg）ELISA检测试剂进行临床特异性及敏感性评价。[方法]制备抗HCV-核心区（Core）抗原四株单克隆抗体，研制出双抗体夹心检测HCV～core抗原酶联免疫检测（ELISA）试剂。三家临床单位收集抗HCV阴性样品3040份，HBsAg阳性100份，抗HIV阳性40份，从10万份抗-HCV筛查出临界样品28人份。[结果]3040份阴性样品检测有3份HCV—cAg阳性，3份经HCVPCR检测有1份阳性，100份HBsAg阳性，40份抗HIV阳性样本检测均为阴性，在10万份抗HCV抗体筛查样品中，选择28份抗-HCV检测临界样品进行HCV—cAg检测，有4份阳性，4份HCV—cAg阳性样品中有3份HCV—RNA检测阳性。[结论]研制的双抗体夹心HCV-核心抗原ELISA试剂具有很好的特异性和灵敏度。     

游离丙型肝炎病毒核心抗原酶联免疫吸附试验不适用于粒细胞集落刺激因子动员的造血祖细胞捐献者的筛查

市售的酶联免疫试验试剂盒抗-丙型肝炎病毒(HCV)核心抗原酶联免疫(ELISA)检测系统(Ortho Clinical Diagnostics, Raritan,NJ),能够检测患者感染HCV后窗口期的HCV核心抗原,即在产生HCV RNA后的1d～2d内检测到HCV核心抗原,灵敏度为94％,总的特异性为99％。由于作者关注的是自体或异体外周血祖细胞(PBPC)产品污染HCV的问题,作者在用HCV抗原试验筛查HPC捐献者时,发现HCV抗原阳性检出率较高。这一发现促使作者进一步研究应用细胞因子动员的患者的HCV抗原检测阳性率高的原因,这种增高是真正病毒复制所致还是检测HCV核心抗原试验方法干扰所致的问题。     

抗原抗体联合检测在提高血液透析患者丙肝感染检出率中的应用

目的探讨开展抗原抗体联合检测在提升血液透析治疗患者丙肝感染检出率中的应用价值。方法收集100例血液透析患者的临床资料,分别采用以下方法测定相关参数:ELISA法检测HCV核心抗原、化学发光法检测抗HCV抗体、RTPCR法检测HCV-RNA,统计各检测项目在同一标本中的阳性表达情况,比较单测抗原、抗体与联合检测3个检测方案对于RNA阳性标本的检出率;同时对抗原阳性标本的抗原水平与其RNA拷贝数进行相关性分析并比较抗体阳性标本中其RNA阳性与阴性组组间抗体水平的区别。结果单测抗原或抗体均会造成血透患者丙肝感染的漏检,对于RNA阳性标本的检测中抗原抗体联合检测的检出率最高(P0.05),抗原阳性标本的抗原水平与其对应标本的RNA拷贝数呈正相关关系(r=0.85,P0.05),而抗体阳性标本中其RNA阳性与阴性组的组间抗体水平的差异不具有统计学意义(P0.05)。结论 HCV抗原抗体联合检测能有效提高血液透析治疗患者丙肝感染的检出率,同时HCV核心抗原检测也可作为抗体阳性患者HCV再次感染的监测指标之一。     

丙型肝炎病毒抗体联合核酸定量检测对丙型肝炎患者的早期诊断价值研究

目的：探讨在丙型肝炎患者早期诊断中丙型肝炎病毒抗体联合丙型肝炎病毒核酸（HCV RNA）检测的临床价值。方法对116例鼓楼医院住院及门诊丙型肝炎患者采用实时荧光定量聚合酶链反应（FQ-PCR）测定HCV RNA、酶联免疫吸附试验（ELISA）和金标法测 HCV抗体、速率法测丙氨酸氨基转移酶（ALT ）。结果116例丙型肝炎患者中,ELISA HCV抗体阳性97例（占83．6％）,HCV RNA阳性85例（占73．3％）,HCV 金标阳性77例（占66．4％）,ALT阳性69例（占59．5％）,HCV抗体和 HCV RNA联合检测总阳性率（指任-指标阳性即为阳性）为100．0％。结论 HCV抗体和HCV RNA联合检测扩大了丙型肝炎检测范围,降低了丙型肝炎的漏诊率,有利于丙型肝炎的早期诊断。     

Performance of an enzyme-linked immunosorbent assay system for antibodies to hepatitis C virus with two new antigens (c11/c7).

We developed an enzyme-linked immunosorbent assay (ELISA) system for antibodies to the hepatitis C virus (HCV), using two new recombinant antigens (c11 and c7) derived from the HCV genome. The performance of this ELISA system (Imucheck HCV Ab) was examined. The CV values for both intra-assay precision and reproducibility of identifying HCV antibody in the panel sera ranged from 3.5% to 6.4%. The blood elements in serum and anticoagulants did not interfere in this ELISA system. The specificity of Imucheck HCV Ab to samples from patients with non-A, non-B (NANB)-type chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma was 93.7%, 93.5%, and 81.4%, respectively. These results are more sensitive than those obtained by the first-generation anti-HCV ELISA system. In the samples from patients with NANB-type acute hepatitis, Imucheck HCV Ab enabled detection of HCV antibodies at an early stage. This system increased the sensitivity for blood donor screening and for monitoring patients with acute hepatitis.     

丙型肝炎病毒总抗原检测用于病程监测的研究

目的探讨丙型肝炎病毒(HCV)总抗原检测方法在丙肝病程监测方面的临床意义。方法对来本院就诊的40位丙肝患者于治疗前、治疗1个月时、治疗3个月时、治疗6个月(停药)时,停药6个月后等不同时期进行采血,收集血清或血浆标本,用抗-HCV检测试剂盒(酶联免疫法)、HCV核酸(RNA)扩增(PCR)荧光定量检测试剂盒、HCV总抗原检测试剂盒(酶联免疫法)进行检测。结果从患者确认感染丙肝到治疗结束抗-HCV检测均呈阳性,而HCV-RNA检测和HCV总抗原检测会随着病程的变化而变化。本次共检测了189例标本(40位患者不同时期标本总例数),其中HCV-RNA阳性51例,该51例阳性标本中,HCV总抗原检测阳性44例,阳性检出率为86.27%;138例HCV-RNA阴性标本,有3例HCV总抗原检测为阳性(2.2%)。2种方法比较,差异无统计学意义(χ2=1.6,P>0.05)。HCV总抗原检测其OD值会随着病程的变化而相应改变,可以较好地反应丙肝患者的病程状况。结论 HCV总抗原检测方法在丙肝病程监测方面具有很好的临床意义,适合在缺少荧光定量PCR检测能力的中小医院使用,可在一定程度上替代HCV-RNA检测,对抗-HCV阳性患者作进一步的验证检测或补充,更好地应用于丙肝患者的病程监测。     

抗-HCV结果分析及灰区范围设置的探讨

目的分析酶联免疫吸附试验(ELISA)筛查丙型肝炎病毒抗体(抗-HCV)结果,探讨实验室ELISA中灰区范围设置的必要性。方法回顾性分析34 942例患者抗-HCV筛查结果,比较抗-HCV与HCV-RNA及临床确诊丙型肝炎患者间的关系;以初筛S/CO值在0.4~2.0范围内的标本为灰区样本,进行不同厂家试剂复检及HCV-RNA检测,探讨设置抗-HCV检测灰区范围的必要性。结果抗-HCV筛查阳性率0.61%;31~50岁阳性率最高;男性高于女性,两者比较差异有统计学意义(P0.05)。S/CO≥10时抗-HCV与HCV-RNA检测结果符合程度高,S/CO≥3.8时抗-HCV与临床确诊丙型肝炎符合程度高。灰区样本的阳性率0.38%,双试剂双孔复检后阳性率0.20%和0.05%。结论抗-HCV筛查阳性率在不同地域、性别及年龄段存在差异;S/CO值越大,HCVRNA阳性率越高,与临床丙型肝炎的确诊符合程度越高,而抗-HCV筛查落在灰区范围的样本应复检并检测RNA,以减少实验室漏检或假阳性结果的产生。     

HCVRNA定量及HCV核心抗原和丙氨酸氨基转移酶结果分析

目的分析丙型肝炎病毒(HCV)RNA定量检测结果与HCV核心抗原及丙氨酸氨基转移酶(ALT)检测结果的相关性。方法收集涪陵监狱卫生所94例住院和门诊患者血清,采用逆转录聚合酶链反应荧光探针法定量检测标本中的HCV RNA,同时采用酶联免疫吸附试验检测HCV核心抗原,采用全自动生化分析仪检测ALT水平。结果 94例样本中HCV RNA阳性率为56.4%(53/94),HCV核心抗原阳性率为53.2%(50/94),经统计学分析,两种方法的阳性率差异无统计学意义(P>0.05),符合率为84.9%(45/53);ALT异常率为62.3%(33/53),随着HCV RNA含量的升高而增加。结论 HCV RNA定量检测结合HCV核心抗原及ALT检测,可帮助临床了解HCV在体内的复制水平及肝脏的炎性反应状态,以指导临床用药及疗效观察。     

Detection of free hepatitis C virus core antigen by enzyme-linked immunosorbent assay is not suitable for screening of granulocyte colony-stimulating factor-mobilized hematopoietic progenitor donors

BACKGROUND: Recent studies have shown that hepatitis C virus (HCV) can be detected in peripheral blood mononuclear cells of patients who are negative for the presence of anti-HCV and serum HCV RNA. The aim of the study was to evaluate the prevalence of HCV viremia in granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood progenitor cell (PBPC) donors by the use of a free HCV core antigen enzyme-linked immunosorbent assay (ELISA). STUDY DESIGN AND METHODS: A total of 28 samples from consecutive PBPC donors that were mobilized with G-CSF, and 13 samples from patients presenting with leukocytosis of greater than 20 x 10(9) per L from other causes, were tested by a free HCV core antigen ELISA. Positive samples were confirmed by use of neutralization assays. The specificity of the assay was studied in 48,911 healthy blood donors negative for the presence of anti-HCV. RESULTS: The free HCV core antigen assay showed a 46.4 percent positivity in PBPC donors mobilized with G-CSF and 61.5 percent in patients exhibiting leukocytosis in the absence of G-CSF treatment. All the samples were found to be false-positive samples, and those related with growth factor treatment did not react when G-CSF was discontinued. Overall specificity by the test in freshly collected blood donor specimens was 99.62 percent. CONCLUSION: Data indicate that the free HCV core antigen ELISA is not a valid test in diagnosing HCV infection in G-CSF-treated PBPC donors. Moreover, false-positive results of this test on blood donors might be indicative of elevated white blood cell numbers. The low specificity of this assay in the PBPC mobilization setting suggests that molecular assays should be the test of choice in the screening of G-CSF-treated donors.     

HLA and hepatitis C virus positive cardiomyopathy

The relationship between HCV (hepatitis C virus) and the susceptibility of cardiomyopathy has been indicated, but the detailed mechanism for close association is still unknown. It is well known that the human leukocyte antigen (HLA) may regulate the development of chronic hepatitis in HCV positive patients. We have analyzed the distribution of HLA class II alleles in Japanese patients with HCV antibody positive dilated cardiomyopathy (DCM) and hypertrophic cardiomyopathy (HCM), and HLA-DPB1*0901 was significantly increased in HCV Ab positive DCM, and the HLA-DRB1*0901-DQB1*0303 haplotype was in HCV Ab positive HCM. These results suggested that molecular mechanism for the development of cardiomyopathy mediated by HCV is different between DCM and HCM.     

Efficacy of HCV core antigen detection during the preseroconversion period

BACKGROUND: The purpose of this study was to compare the performances of HCV core antigen (HCV Ag) testing with HCV RNA detection during the preseroconversion period. STUDY DESIGN AND METHODS: Six HCV antibody (HCV Ab)-negative and HCV RNA-positive blood samples from 6 donors and 135 serial samples from 28 patients who had undergone hemodialysis, collected a mean of 90 days before the detection of HCV Ab, were tested by ELISA for the detection of HCV Ag and by PCR to quantify HCV RNA. RESULTS: Five of the six donors were positive for HCV Ag. The donor with a negative HCV Ag test had the lowest viral load. In the hemodialysis patients, the 43 first specimens of the series were HCV RNA negative. Of the 92 specimens that were HCV RNA positive, 81 (88%) were positive for HCV Ag. Among the 74 samples with more than 10(5) RNA copies, 71 (96%) were HCV Ag positive. Average time from first viremic bleed to first HCV Ag-positive bleed was estimated at 2.0 days and that to first HCV Ab-positive bleed at 50.8 days. CONCLUSION: HCV Ag testing permits the detection of an HCV infection about 1.5 months earlier than the HCV Ab screening tests and an average of only 2 days later than quantitative HCV RNA detection in individual specimens.     

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.