维持性血液透析患者并发病毒性肝炎的临床研究

目的 了解维持性血液透析患病毒性肝炎的感染率及其有关因素。方法 用酶联免疫法(ELISA)检测53例乙型肝炎病毒(HBV)标志物、丙型肝炎病毒(HCV)抗体，逆转录一套式PCR法检测HCV—RNA。回顾分析53例维持性血液透析患的临床资料。结果 53例维持性血液透析患肝炎病毒感染率分别为乙型肝炎病毒(HBV)22．6％、丙型肝炎病毒(HCV)41．5％、HBV／HCV总感染率49．1％，HCV感染组、非感染组在输血次数、透析年限的差异有显性，而HBV感染组、非感染组在输血次数、透析年限的差异无显性。结论 病毒性肝炎仍是血液透析(HD)的主要并发症之一，其中以HCV的发生率最高。严格消毒措施，血源筛选，减少输血，对减少透析中肝炎感染至关重要。     

肝移植受体病肝组织中丙型肝炎病毒的检出及其临床意义

目的：检测成人肝移植受体丙型肝炎病毒(HCV)感染状态，探讨监测肝移植受体术后丙型肝炎复发的意义。方法：收集本移植中心器官移植受体摘除肝脏标本52例，采用Tordji-22和NS32种单克隆抗体进行免疫组织化学检测。结果：所有病例术前血清抗HCV检测均阴性。52例肝移植受体肝脏组织中20例(38．46％)肝细胞内有不同程度的HCV抗原存在。上述2种单克隆抗体的灵敏度及阳性表达方式略有不同，Tordji-22以肝细胞核表达为主，NS3以肝细胞胞质细颗粒状阳性为主。原发性肝癌肝移植组中HCV阳性细胞主要为癌旁肝细胞，少数癌细胞胞质也呈弱阳性。结论：在接受肝移植的慢性终末期肝病或原发性肝癌患者中，有相当病例受体肝脏有HCV感染。由于肝移植后应用大剂量皮质激素及免疫抑制药物可促进HCV病毒在移植肝的肝细胞内复制，导致丙型肝炎复发，明确肝移植受体肝脏HCV感染状态，对肝移植后病情监测、制定治疗方案具有指导意义。     

丙型肝炎病毒RNA含量与抗-HCV及ALT的关系

目的 了解丙型肝炎病毒核糖核酸(HCV RNA)与丙型肝炎病毒抗体(抗-HCV)及丙氨酸氨基转移酶(ALT)的关系.方法 144例在本院住院和门诊同时检测HCV RNA、抗-HCV及ALT的患者,用荧光定量逆转录聚合酶链反应(FQ-RT-PCR)法检测标本中的HCV RNA,同时采用酶联免疫吸附试验(ELISA)法检测抗-HCV,用全自动生化检测仪测定ALT水平.结果 144例标本中HCV RNA高于上限值34例(23.6%),抗-HCV阳性44例(30.6%),二者有很好的相关性.有46例(31.8%)存在ALT异常,而ALT异常率与HCV RNA含量有一定的比例关系.结论 HCV RNA含量与抗-HCV同时检测可提高临床对丙型肝炎的诊断.HCV RNA是反映HCV复制的可靠指标,结合ALT结果可帮助临床了解HCV在体内的复制状况及肝脏的炎性反应状态.FQ-RT-PCR法检测HCV RNA具有非常好的临床应用价值.     

两种荧光定量法检测HCV RNA结果的比较

目的建立双探针荧光定量HCV RNA检测方法,分析其定量检测HCV RNA的灵敏度和特异性以及HCV RNA含量与患者病情的相关性。方法选取100例抗HCV抗体阳性患者,包括慢性肝炎患者45例、肝硬化患者30例、肝癌患者25例的样本,用双探针荧光定量法和两种商品荧光定量试剂方法进行HCV RNA检测,另观察近期196例抗HCV抗体阳性和48例抗HCV抗体阴性样本荧光定量PCR HCV RNA含量与肝功能的相关关系。结果荧光定量双探针法阳性率为93％（93／100）;两种商品荧光定量试剂方法阳性率分别为84％（84／100）和79％（79／100）,经卡方检验差异有统计学意义（P〈0．05）。196例中抗HCV与HCV RNA阳性一致率为57．1％（116／196）,其中伴肝功能改变49．0％（96／196）。HCV RNA含量与AST和ALT异常程度无显著相关性（P〉0．05）,相关系数分别为0．4293及0．3899。HCV RNA高拷贝数患者肝功能异常率为85．2％（69／81）,低拷贝数者肝功能异常率为40．9％（47／115）,经卡方检验,X^2=38．63,差异有统计学意义（P〈0．001）。结论双探针荧光定量提高了HCV RNA检测的灵敏度和特异性,HCV RNA含量与患者肝功能损伤程度无明显相关性。     

Detection of hepatitis C virus (HCV) RNA in paraffin embedded tissue sections of human liver of non-A, non-B hepatitis patients by in situ hybridization.

Localization of hepatitis C virus (HCV) RNA was investigated by non-radioactive in situ hybridization in human liver specimens of chronic non-A, non-B (NANB) hepatitis patients who were seropositive for antibodies to HCV (anti-HCV). For in situ hybridization, T-T dimerized synthetic oligodeoxynucleotide probes were used and DNAs hybridized in situ were detected immunohistochemically using specific antibodies against T-T dimer. The data demonstrates that HCV-RNA was localized in the cytoplasm of hepatocytes in human liver biopsies obtained from the patients with chronic NANB hepatitis seropositive for anti-HCV.     

Enzyme-linked immunosorbent assay for antibodies against the capsid protein of hepatitis C virus with a synthetic oligopeptide

An enzyme-linked immunosorbent assay was developed for the determination of antibodies against the putative capsid protein of hepatitis C virus (HCV). A 36-mer oligopeptide with a sequence of RRGPRLGVRATRKTSERSQPRGRRQPIPKVRRPEGR (CP9) was synthesized; it was selected on the translation product of the presumptive HCV core gene, because of a high local hydrophilicity and excellent conservation by different HCV strains. The synthetic peptide was immobilized on a solid-support to capture antibodies directed to CP9 (anti-CP9) in test sera, which were detected by Fab' fragments of monoclonal anti-human IgG/gamma labeled with horseradish peroxidase. The specificity of anti-CP9 was confirmed by absorption tests. Anti-CP9 was detected in 13 (68%) of 19 patients with sporadic acute non-A, non-B (NANB) hepatitis and in 15 (83%) of 18 patients with post-transfusion acute NANB hepatitis. In 7 cases of acute NANB hepatitis who were followed, anti-CP9 developed earlier than antibodies against HCV (anti-HCV) detectable by a commercial assay kit. Among patients with chronic NANB liver diseases, anti-CP9 was detected in 103 (77%) of 133 with chronic hepatitis, 70 (62%) of 113 with liver cirrhosis and 31 (76%) of 41 with hepatocellular carcinoma. Anti-CP9 and anti-HCV overlapped in 175 (54%) among 324 cases of acute or chronic NANB liver diseases; 58 (18%) were positive only for anti-CP9 while 49 (15%) were positive only for anti-HCV. HCV RNA was detected, by amplifying HCV cDNA with polymerase chain reaction, in 10 of 11 sera positive only for anti-CP9. Among sera from 606 blood donors, 21 were positive only for anti-CP9. HCV RNA was detected in 5 (24%) of them, all of which had A492 values greater than 0.600 in ELISA for anti-CP9. Based on these results, anti-CP9 would complement anti-HCV for the diagnosis of HCV infection and contribute toward further decreasing posttransfusion NANB hepatitis.     

库血HCV-RNA的检测分析

目的 为探索提高库血丙型肝炎病毒(HCV)的检出率。方法 采用逆转录套式聚合酶链反应(RT-PCR)技术，检测4530份抗-HCV阴性的库血HCV-RNA。结果 4530份库血中HCV-RNA阳性55份，阳性率1．2l％．其中个体献血者血液2110份，HCV-RNA阳性40份，阳性率1．90％；无偿献血者血液2420份，HCV-RNA阳性15份，阳性率0．62％。结论 RT-PCR可用于检测库血HCV-RNA，以减少因输血造成HCV感染。     

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.     

丙型肝炎病毒感染的献血者10年追踪观察

目的　追踪观察发生HCV感染的献血者的疾病进程和健康状况。方法　从 1993年 10月～ 2 0 0 4年 2月 ,对 30名HCV感染的献血者定期抽取血液标本 ,观察所得到的 4 4 2份系列标本的血清ALT、抗 HCV和HCV RNA的动态变化 ,并进行HCV分型测定 ,重点分析了其中 10名自愿者肝组织的病理检查结果和血清ALT、抗 HCV与HCV RNA的动态变化。结果　① 30名HCV感染者系列标本中 ,ALT异常率 37.6 % (16 6 / 4 4 2 ) ;抗 HCV阳性率97.1% (42 9/ 4 4 2 ,ELISA法 ) ;HCV RNA阳性率 74 .9% (331/ 4 4 2 )。② 10名HCV感染的献血者 10年系列血清中抗 HCV大多数持续存在 (94 4 % ,15 1/ 16 0 ) ,其HCV RNA时有间隙性阴性 ,阳性率 6 7 5 % (10 8/ 16 0 )其血清ALT水平异常率 33 6 % (5 0 / 14 9)。③HCV分型测定 :HCVⅡ / 1b型占 85 % (2 2 / 2 6 ) ,Ⅲ / 2a型占 15 % (4/ 2 6 )。④ 10名HCV感染者肝组织检查显示均为轻度慢性肝炎。结论　肝组织的病理变化同血清抗 HCV、HCV RNA和ALT的异常有明显相关性。虽然HCV感染后 10年的疾病进程大多是缓慢的 ,但应采取适当治疗措施以控制慢性肝炎的发展。     

HCV-IgG、HCV-cAg与HCV-RNA对丙型肝炎诊断的评价

目的评价丙型肝炎病毒核心抗原(HCV-cAg)、丙型肝炎病毒抗体(HCV-IgG)及丙型肝炎病毒RNA(HCV-RNA)3种检测方法在丙型肝炎实验室诊断中的意义。方法收集84例丙型肝炎疑似患者和87例健康对照者血清,采用ELISA法检测HCV-cAg和HCV-IgG,实时荧光定量聚合酶链式反应法(RT-PCR)检测HCV-RNA。结果 84例丙型肝炎疑似患者中HCVIgG阳性率为84.5%,HCV-cAg阳性率为13.1%,HCV-RNA阳性率为52.4%;71例HCV-IgG阳性患者中HCV-RNA阴性35例,假阳性率为49.3%,11例HCV-cAg阳性患者中HCV-RNA阴性5例,假阳性率为45.5%;44例HCV-RNA阳性的丙型肝炎确诊患者中HCV-IgG假阴性率为18.2%,HCV-cAg假阴性率为86.4%;HCV-cAg和HCV-IgG联合检测的假阴性率为13.6%,真阳性率为100.0%。结论 HCV-cAg和HCV-IgG在丙型肝炎的实验室诊断中均存在一定的假阴性和假阳性,将二者联合检测或在必要时与HCV-RNA三者联合检测可降低漏诊率。     

丙肝患者治疗前后HCV RNA与抗-HCV及丙氨酸氨基转移酶水平的分析

目的分析丙型肝炎患者治疗前血清HCV RNA、抗-HCV和丙氨酸氨基转移酶(ALT)水平及治疗后HCV RNA和ALT水平的变化规律。方法对87例丙型肝炎患者血清进行实时荧光定量法(PCR)检测HCV RNA、ELISA法检测抗-HCV和酶速率法检测ALT浓度水平,并对所得数据进行统计学分析。结果丙型肝炎患者血清抗-HCV抗体滴度显著高于健康对照组,大部分患者的ALT水平均有显著性升高;HCV RNA含量与ALT水平具有显著相关性(r=0.673,P0.01),而与抗-HCV的S/OD值无显著相关性(r=0.122,P0.05);在治疗早期,患者HCV RNA含量在第2天显著性下降,而ALT浓度呈一过性上升。结论在HCV诊断与疗效观察中,血清HCV-RNA、抗-HCV和ALT指标各有利弊,3者有机结合在正确诊断和预测肝脏损伤及早期疗效观察中具有重要的临床意义。     

天津市某三甲医院维持性血液透析患者血源性病毒感染情况分析

【摘要】目的了解血液透析患者乙型肝炎病毒（hepatitisBvires,HBV）、丙型肝炎病毒（hepatitisCvirus,HCV）、人类免疫缺陷病毒humanimmunodeficiencyvirus,HIV）和梅毒螺旋体（treponemapal—lidum,TP）的感染情况。方法应用酶联免疫法检测449例维持性血液透析患者血清中HBV五项、抗-HCV、抗-HIV和TP抗体．应用PCR-荧光探针法检测HBsAg阳性的透析患者血清中HBV—DNA含量及抗-HCV阳性的透析患者血清中HCV—RNA含量,并进行统计学分析。结果449例透析患者HB．sAg、HBsAb、抗-HCV、TP抗体的阳性率分别为2．90％、20．71％、8．69％、1．78％,抗-HIV检测结果均为阴性。其中,HBsAg阳性患者中,HBV-DNA阳性率为46．15％,DNA拷贝数为1．1×104—3．7×10^7 IU／mL;抗-HCV阳性者中,HCV-RNA阳性率为58．97％,RNA拷贝数为8．1×10^3～1．5×10^7IU／mL。≥60岁和〈60岁两组患者HBsAg、抗-HCV阳性率比较,差异均无统计学意义（P均〉0．05）。结论血液透析患者HBV、HCV感染率相对国外较低,但感染现象在各年龄段普遍存在。TP感染情况与国内外报道相似,应加强管理防护。     

抗-HCV联合TMA定性检测HCV—RNA在维持性血液透析患者丙型肝炎病毒感染早期诊断中意义的探讨

目的研究维持性血液透析患者丙型肝炎病毒（HCV）感染的血清学诊断方法,探讨抗．HCV联合HCV-RNA检测在维持性血液透析患者HCV感染早期诊断中的意义,以期获得维持性血液透析患者HCV感染早期诊断的可靠方法。方法选择深圳市第二人民医院血液透析中心183例维持性血液透析患者为研究对象,分别使用国产和进口HCV抗体试剂盒检测抗-HCV,使用TMA法定性检测HCV—RNA,荧光定量PCR法定量检测HCV-RNA,比较不同产品及诊断方法对HCV的检出率,评价ALT变化与HCV—RNA载量的关系,评价抗-HCVS／CO值与HCV-RNA载量的关系。结果国产和进口试剂检测抗-HCV的检出率均为7．1％（P=1．000）;TMA法定性检测HCV—RNA的检出率为8。7％,免疫荧光定量PCR法的检出率为5．5％,但两者之间差异有统计学意义（x^2=87．537,P=0．000）;HCV-RNA定性检测比检测抗．HCV的检出率高,两者的差异有统计学意义（x^2=81．531,P=0．000）;联合抗-HCV和HCV-RNA定性检测结果HCV阳性共19例,检出率为10．4％,与单独检测抗．HCV比较差异有统计学意义（P=0．031）,但与单独定性检测HCV—RNA比较差异无统计学意义（P=0．250）。HCV．RNA的载量和ALT的变化无相关性（r=0．189,P=0．536）;抗-HCV初筛的S／CO值与HCV—RNA载量无相关性（r=0．174,P=0．569）。结论HCV．RNA定性检测较抗．HCV检测能缩短维持性血液透析患者HCV感染检出的窗口期,有利于早期诊断。HCV—RNA定性检测能较临床现行的HCV．RNA免疫荧光定量检测显著提高HCV感染的检出率。联合抗-HCV和TMA法定性检测HCV—RNA既能缩短HCV感染的“窗口期”,也能显著提高HCV感染的检出率,可避免漏检处于血清转换期或慢性病毒携带或既往感染的“隐性”患者,值得临床推广应用。ALT的变化和HCV载量无明显相关性,在血液透析患者中辅助早期诊断HCV感染的作用较小。     

Antibody pattern to hepatitis C virus antigens in patients with acute and chronic liver diseases

The present paper describes the antibody pattern in the early stages of hepatitis C virus (HCV) infection and compares it with that obtained in chronic carriers. Samples were tested for anti-HCV from 470 consecutive patients admitted to gastroenterology services at three Havana hospitals, with serological and biochemical signs of non-A, non-B hepatitis, including those referred from three Havana blood banks because of a positive result in anti-HCV in pre-donation screening. Four anti-HCV enzyme immunoassay (EIA) systems and two supplementary assays were used for antibody detection. These results show that antibodies to viral core antigens are the first and commonest among Cuban patients infected with HCV, and generally, they can reveal the infection before the supplementary tests do. LiaTek HCV has proved to be a good tool for the confirmation of enzyme-linked immunosorbent assay (ELISA) positive samples.     

丙型肝炎病毒基因分型与治疗效果关系的研究

目的 分析丙型肝炎病毒（HCV）基因型与利巴韦林联合聚乙二醇干扰素α-2a治疗效果的关系,为临床抗HCV治疗提供依据.方法 选择179例慢性丙型肝炎初治患者进行利巴韦林联合聚乙二醇干扰素α-2a的标准方案进行治疗,采用实时荧光定量聚合酶链反应（PCR）进行HCV-RNA定量检测,采用基因测序检测进行HCV基因分型.治疗后评价持续病毒学应答（SVR）.结果 179例丙型肝炎患者中,HCV 1b型占79.3%,HCV 2a型占8.4%,HCV 3b型占5.0%,HCV 6a型占2.2%,HCV 1b、2a混合型占1.1%,其他型占4.0%.HCV 2a型患者获得SVR率明显高于HCV 1b型,差异有统计学意义（χ^2=4.956,P=0.026）.结论 该院HCV基因型主要为HCV 1b和HCV 2a型,HCV 1b型患者对标准治疗方案的疗效较2a型差.     

Risk factors in hepatitis C virus-infected blood donors

Risk factors of parenteral and nonparenteral exposure to hepatitis C virus (HCV) infection were studied in 125 blood donors in The Netherlands who were positive for anti-HCV on enzyme-linked immunosorbent assay (ELISA). Risk factors were related to confirmatory test results of four-antigen recombinant immunoblot assay (4-RIBA) and polymerase chain reaction (PCR) of the HCV 5' untranslated region. Twelve (10%) of the 125 anti-HCV C100 ELISA-positive blood donors were 4-RIBA positive. Eleven (92%) of 12 4-RIBA-positive blood donors were PCR positive, and all 113 remaining 4-RIBA-negative or -indeterminate donors were PCR negative. Eleven (92%) of 12 4-RIBA-positive blood donors had a risk factor of parenteral exposure, as compared to 17 (15%) of 113 4-RIBA-negative or -indeterminate donors. The prevalence of confirmed HCV infection among Amsterdam blood donors is calculated at 0.04 percent; parenteral exposure appears to be the major risk factor for HCV infection.     

Lichen planus and hepatitis C virus in the Northern Kyushu region of Japan

Abstract. Oral lichen planus (OLP) is a common oral disorder that manifests a mucosal reaction to a variety of aetiological factors, including liver disorder. This study investigated the relationship between OLP and hepatitis C virus (HCV) infection by studying the prevalence of hepatitis B and C virus infection or liver disease in 45 patients with OLP in the Northern Kyushu region of Japan where the prevalence of HCV infection is the highest in the country. Serum hepatitis B virus surface antigen (HBsAg) was positive in only four patients. Serum anti-HCV or serum HCV RNA was positive in 28 (62%) and 27 (60%) of 45 OLP patients, respectively. The majority (35 of 45, 78%) of OLP patients suffered from liver disease, including chronic hepatitis C (22/45, 49%), HCV-related liver cirrhosis (two), and HCV-related hepatocellular carcinoma (two). These results suggest that HCV is a major cause of OLP.     

Comparative evaluation of Abbott and Murex antibody assays and Roche Amplicor PCR for diagnosis of human hepatitis C

Of 135 serum samples from 135 patients suspected of hepatitis C virus (HCV) infection, 67 were detected by Abbott IMX antibody assay, 89 by Murex anti-HCV (version III), and 47 by Roche Amplicor polymerase chain reaction (PCR). Furthermore, 44 of the 62 positive serum samples by both Abbott and Murex antibody assays, 2 of the 27 positive samples by Murex antibody assay only, none of the 5 positive samples by Abbott antibody assay only, and one of the 43 negative samples by both Abbott and Murex antibody assays had measurable HCV RNA by Roche Amplicor PCR, suggesting active hepatitis C viremia. Whereas Abbott and Murex antibody assays were in agreement with each other in 103 of the 135 serum samples tested, they showed discrepancy with regard to the other 32. Despite generating a small percentage of false positives, Abbott and Murez antibody assays are useful in monitoring serum antibody levels of the past or continuing hepatitis C virus infection. Abbott IMX appears to be more specific than Murex anti-HCV (version III). The use of Roche Amplicor PCR provides a means of revealing active hepatitis C viremia, and helping clarify antibody indeterminate serum samples.