抗-HCVELISA间接法与双抗原夹心法试剂的应用对比评价

目的:对比研究国产抗-HCV ELISA间接法和双抗原夹心法试剂的检测效能,探讨血站抗-HCV检测模式。方法:选择1种双抗原夹心法酶联免疫试剂、2种间接法酶联免疫试剂分别检测34 593名无偿献血者血样,采用重组免疫印迹试验(RIBA)对其中有反应性的44份标本进行确认,并用BBI血清盘对这2种检测试剂进行考核评价。结果:科华、新创和万泰3个厂家的抗-HCV有反应性及灰区标本经RIBA实验确证后,假阳性率分别为31%、63%、13%。万泰双抗原夹心法与间接法相比,增加了反应强度、降低了假阳性率且缩短了窗口期。结论:为确保血液质量,血筛实验室应选择灵敏度和特异性双优的试剂,采用间接和双抗原夹心2种不同的ELISA试剂检测HCV抗体优于2种间接ELISA试剂,不仅提高抗-HCV有反应性标本的检出率,而且减少血液因假阳性造成的浪费。     

血站实验室献血者HCV抗体检验报告新模式探讨

目的评估血站日常抗-HCV间接法酶联免疫吸附实验检测结果的准确性,探讨如何转换抗-HCV报告流程,保护献血者的积极性及珍贵的血液资源。方法使用上海科华间接法丙型肝炎试剂盒(试剂A)、北京万泰间接法丙型肝炎试剂盒(试剂B)对116份标本进行间接法酶联免疫检测,测定结果为阳性或灰区的标本,使用确认试剂进一步检测,确定其真实结果,统计本地区丙型肝炎确认结果与双试剂检测结果的对应分布情况。两种抗-HCV试剂对116份标本的检测采用配对χ2检验,对两种试剂的检验结果采用一致性检验。结果两种间接法试剂对116例标本的检测结果,差异具有统计学意义(P=0.04),而两种试剂一致性较差,双试剂强阳性标本、弱阳性标本的假阳性率分别为0、35.7%;两种试剂的单试剂阳性标本、灰区标本的假阳性率分别为A试剂:94.3%、100%;B试剂:84.2%和88.9%。结论间接法试剂假阳性率较高,特异性较差,国产间接法试剂之间存在较大差异,应对原丙型肝炎报告方式进行修改,对弱阳性标本应进行进一步的确证实验以保护献血者积极性。     

乙型和丙型肝炎重叠感染的状况研究

本文主要对1992年1月-2001年10月我院收治符合2000年9月全国病毒性肝炎防治方案的673例乙型肝炎患者进行了HBV、HCV病毒标志物的检测和回顾性分析。 1 临床资料 673例乙型肝炎患者血清抗-HCV阳性率为18.72％,其中重型肝炎阳性率最高(46.34％),慢性肝炎和肝硬变次之(22.52％、16.75％)。各临床类型抗-HCV阳性率有显     

4375例输血前4项检测结果分析

目的：了解患者输血前传染性病原体感染状况。方法：运用酶联免疫吸附试验法和甲苯胺不加热血清试验法对4375例住院患者进行HBsAg、抗-HCV、抗-HIV、梅毒等“输血前4项”检测。结果：4375例患者检测总阳性率505例（11．54％），其中HBsAg阳性439例（10．03％）、抗-HCV58例（1．32％），抗HIV阳性2例（0．05％）、RPR6例（0．14％）。结论：进行输血前4项检测有利于患者的治疗及医院感染的预防，减少因输血而引起的医疗纠纷。     

国产某丙型肝炎病毒抗体试剂中吸光度/临界值的作用

目的研究某国产抗-HCV试剂样本吸光度／临界值（S／Co）的意义，探讨国产试剂检测抗-HCV的初步程序和标准。方法采用某国产试剂通过ELISA法检测295000例无偿献血者的抗-HCV，对其阳性反应的样本采用Ortho抗-HCV试剂通过ELISA法进行检测。Ortho试剂阳性反应的样本进行HCV重组免疫印迹实验（RIBA），并对其中的106例Ortho试剂阳性反应的样本进行核酸检测。结果在295000例样本中，某国产试剂抗-HCV阳性反应为681例，阳性率为0．23％。某国产试剂阳性反应的样本经Ortho试剂检测后阳性反应为367例，符合率为53．8％，其中Ortho试剂S／Co值≥3．8为66．2％。在Ortho试剂S／Co值≥3．8的样本中，某国产试剂S／Co值≥8．0的占94．2％；而Ortho试剂S／Co值〈3．8的样本中，某国产试剂S／Co值〈8．0的占99．2％。367例Ortho阳性反应样本中，RIBAHCV阳性223例，阳性率为60．8％；在国产试剂S／Co值≥8．0的样本中，RIBA阳性率为95．7％；而同一种国产试剂S／Co值〈8．0的样本中，RIBA阳性率为2．2％。106例Ortho试剂阳性反应样本中核酸检测阳性为42例。结论国产抗-HCV试剂在检测无偿献血者样本中存在较高的假阳性率，一定程度上造成了血源和血液的浪费。某国产试剂S／Co值≥8.0和Ortho试剂S／Co值≥3．8具有一定的相关性，在实验室检测抗-HCV上具有一定诊断价值。     

重型肝炎患者抗-HCV检测结果分析

HCV感染与慢性肝炎、肝硬化及肝癌的关系已渐明确,但HCV在重型肝炎发病中的作用目前少见报道,为了了解HCV感染与重型肝炎的联系,我们对55例重型肝炎患者进行了血清抗-HCV检测,现报告如下: 材料与方法 一、病例 55例均系我院1991年11月～1993年7月住院患者,其中男51例,女4例,年龄11～60岁,平均42岁。根据1990年上海病毒性肝炎会议制订的诊断标准,符合急性重型肝炎5例,亚急性重型肝炎10例,慢性重型肝炎40例。 二、方法 抗HCV检测采用酶联免疫法(EIA),试剂由上海长征医学科学有限公     

2006～2012年天津市住院精神病患者病毒性肝炎感染状况调查

目的调查分析天津市住院精神病患者病毒性肝炎感染情况。方法2006年1月-2012年12月住院精神病患者19994例,采用酶联免疫法检测血清HBsAg、抗-HBs、HBeAg、抗-HBe、抗-HBc、抗-HCV、抗-HEV—IgM。结果HBsAg阳性627例,阳性率3．14％;抗．HCV阳性102例,阳性率0．51％;抗-HEV—IgM阳性24例,阳性率0．12％;HbsAg＋抗-HCV阳性5例,阳性率0．03％;HbsAg＋抗-HEV—IgM阳性2例,阳性率0．01％;HbsAg＋抗-HBe＋抗-HBc阳性（小三阳）385例,阳性率1．93％;HBsAg＋HBeAg＋抗-HBc阳性（大三阳）115例,阳性率0．58％;Hb-sAg＋抗．HBc阳性127例,阳性率0．64％。结论住院精神病患者病毒性肝炎感染率高,应根据精神病住院患者的特点,加强病房管理,防止病毒性肝炎院内感染及长期用药加重肝损伤。     

两种血吸虫病免疫诊断试剂盒临床诊断特异性的评价

目的评价胶体染料试纸条(DDIA)和酶联免疫吸附试验(ELISA)两种试剂盒在血吸虫病临床诊断中的价值。方法采集非血吸虫病流行区的华支睾吸虫、并殖吸虫病人、健康正常人群血清和乙肝病人血清,按照单盲试验进行DDIA和ELISA检测。结果 DDIA和ELISA两种试剂盒,检测186份健康献血员的特异性分别为98．92％和99．6％,检测35例并殖吸虫病人的交叉反应率为42．86％和17．14％,检测31例华支睾吸虫病人的交叉反应率为9．68％和6．45％,检测159 份乙肝病人假阳性率分别为1．26％和5．03％;DDIA试剂盒与并殖吸虫病人的交叉反应显著高于 ELISA试剂盒(X2=5．510,P=0．019),DDIA试剂盒检测乙肝病人和健康正常人群的阳性率差异无显著性(X2=0．025,P=0．875),ELISA试剂盒检测乙肝病人的阳性率显著高于健康正常人群(X2 =6．814,P=0．009),其阳性率是健康正常人群的9．36倍。结论在血吸虫病现场筛查和医院门诊检查血吸虫病时,DDIA试剂盒优于ELISA试剂盒。     

ELISA法快速检测慢性肝病患者血清中TIMP-1

目的 应用酶联免疫吸附方法（ELISA）快速检测慢性肝病患者血清中基质金属蛋白酶组织抑制剂-1（TIMP-1），了解TIMP-1血清学检测与肝纤维化病程进展程度。方法 以抗人TIMP-1单克隆抗体为包被抗体，15％小牛血清为封闭液，辣根过氧化物酶（HRP）标记的羊抗人IgG作为酶结合物建立检测人血清TIMP-1 ELISA法，用于临床检测399例肝病患者血清和健康人血清105例。结果 临床检测表明肝硬化患者TIMP-1检出阳性率为76．9％，明显高于慢性肝炎（40．7％）和急性肝炎（19．4％）（P〈0．005），并随着纤维化程度加重而升高（P〈0．01）。结论 血清中TIMP-1的变化可作为肝纤维化较为有用的诊断指标。ELISA法具有较高的敏感性、特异性和快速性，试剂用量少，且不需复杂的仪器设备，加之实验程序简单，甚适于医院及基层医疗单位应用。     

2008-2010年北京协和医院住院患者弓形虫感染的血清流行病学调查

目的 了解北京协和医院住院患者弓形虫感染的情况.方法 收集2008-2010年期间北京协和医院住院和门诊患者的血样,用酶联免疫吸附试验(ELISA)检测血清抗弓形虫IgG抗体,计算住院患者血清弓形虫抗体的阳性率,并按科室和疾病分别统计分析患者弓形虫抗体的阳性率.结果 住院组和门诊组患者的弓形虫抗体阳性率分别为2.22%...     

Estimation of IgG subclasses of anti-HCV (C100-3) in non-A,non-B fulminant,acute and chronic hepatitis and it’s significance

To determine whether identification of subclasses of anti-HCV IgG would help distinguish between acute and fulminant hepatitis, we assayed serum IgG subclasses of anti-HCV (C100-3) in non-A, non-B hepatitis. Anti-HCV IgG was restricted to two subclasses in different diagnostic conditions; anti-HCV IgGl and anti-HCV IgG3. Anti-HCV IgGl was the main subclass in acute hepatitis (AH), and was positive in all the cases and only one case was positive for anti-HCV IgG3. Anti-HCV IgG3 was the dominant subclass in fulminant hepatitis (FH). Out of the total 12 cases of FH, who were positive for either anti-HCV IgG or for any of it’s subclasses, in 10 cases (83%), anti-HCV IgG3 had higher optical density (OD) values than anti-HCV IgGl and in 6 cases (50%) was the only subclass of anti-HCV IgG being positive. In 5 cases with FH, anti-HCV IgG3 became positive even when anti-HCV IgG was negative. These findings form the basis of a new observation and are likely to be helpful in the diagnosis of non-A, non-B fulminant hepatitis. This study was supported by a Grant-in-Aid for Scientific Research from viral Hepatitis and Research Foundation of Japan, 1991.     

Minimal transmission of HIV despite persistently high transmission of hepatitis C virus in a Swedish needle exchange program

The aim of this study was to examine the prevalence and incidence of HIV and hepatitis B and C (HBV and HCV) among injecting drug users in a Swedish needle exchange programme (NEP) and to identify risk factors for blood-borne transmission. A series of serum samples from NEP participants enrolled from 1997 to 2005 were tested for markers of HIV, HBV and HCV (including retrospective testing for HCV RNA in the last anti-HCV-negative sample from each anti-HCV seroconverter). Prevalence and incidence were correlated with self-reported baseline characteristics. Among 831 participants available for follow-up, one was HIV positive at baseline and two seroconverted to anti-HIV during the follow-up of 2433 HIV-negative person-years [incidence 0.08 per 100 person-years at risk (pyr); compared to 0.0 in a previous assessment of the same NEP covering 1990-1993]. The corresponding values for HBV were 3.4/100 pyr (1990-1993: 11.7) and for HCV 38.3/100 pyr (1990-1993: 27.3). HCV seroconversions occurred mostly during the first year after NEP enrolment. Of the 332 cases testing anti-HCV negative at enrolment, 37 were positive for HCV RNA in the same baseline sample (adjusted HCV incidence 31.5/100 pyr). HCV seroconversion during follow-up was significantly associated with mixed injection use of amphetamine and heroin, and a history of incarceration at baseline. In this NEP setting, HIV prevalence and incidence remained low and HBV incidence declined because of vaccination, but transmission of HCV was persistently high. HCV RNA testing in anti-HCV-negative NEP participants led to more accurate identification of timepoints for transmission.     

Detection of serum antibody against hepatitis C virus in patients with hepatitis and liver diseases]

Antibody against hepatitis C virus (anti-HCV) was tested in 658 cases of hepatitis and liver diseases with ELISA, ninety of these cases were positive, with a total infection rate of 13.68% (90/658). The positive rate of anti-HCV was highest in patients with chronic severe hepatitis (33.78%) and CAH accompanied by cirrhosis of liver(31.58%). The infection rate in other types of hepatic diseases in order of frequency was as follows: fulminant hepatitis (18.18%), CAH without cirrhosis (15.13%), subacute severe hepatitis (13.43%), CPH (5.88%), primary hepatocellular carcinoma (3.85%), and acute hepatitis (2.42%). Serological markers of HBV infection were detectable concomitantly in 77 of the 90 cases who were anti-HCV positive, but there was no evidence of mutual inhibition of viral replication. There was neither appreciable difference in the level of hyperbilirubinemia in cases of hepatitis with or without anti-HCV, nor significant diversity in the number of death between cases of severe hepatitis with and without anti-HCV.     

Hepatitis C virus infection in Italian intravenous drug users: epidemiological and clinical aspects

Abstract: The epidemological and clinical features of hepatitis C virus infection have been evaluated in a cohort of 227 intravenous drug users enrolled at a drug dependence treatment center in the Veneto area in 1992–1993 and followed periodically. Hepatitis C virus infection was detected using second-generation anti-HCV ELISA in 171 (75%) subjects at enrollment. Anti-HCV seropositivity correlated with: a) the duration of drug abuse: 91% of intravenous drug users injecting for more than 8 years were seropositive as compared to 40% of those with a history of abuse lasting 4 years or less, p<0.001; b) sharing of injection equipment: 85% anti-HCV positive intravenous drug users had shared at some time as compared to 64% seronegative subjects, p<0.001; c) seropositivity for immunodeficiency virus infection: 25% anti-HCV positive intravenous drug users were coinfected as compared to 3.5% anti-HCV negative, p<0.001; d) markers of ongoing (two cases) or previous hepatitis B virus infection were detected in 62% of anti-HCV positive but in 21% of anti-HCV negative cases, p<0.01. Two initially anti-HCV negative intravenous drug users seroconverted during follow up giving an incidence rate of hepatitis C virus infection of 6.2 per 100 person-years. During the survey abnormal alanine aminotransferase levels were detected in 75% anti-HCV positive but in 24% anti-HCV negative cases (p<0.001), with significantly higher levels in the former. These findings suggest that the circulation of hepatitis C virus among intravenous drug users has been decreasing in recent years, although new infections still occur. In agreement with the high rate of chronicization of parenterally transmitted hepatitis C, the majority of anti-HCV positive subjects had biochemical features of associated liver disease.     

Outcome of acute symptomatic non-A,non-B hepatitis: a 13-year follow-up study of hepatitis C virus markers

ABSTRACT— Thirty-nine of 61 prospectively followed patients who had had acute non-A, non-B hepatitis in 1978 were clinically reexamined in 1991 and tested for antibodies to hepatitis C virus (anti-HCV) with a second generation ELISA and RIBA and for HCV RNA by PCR. Acute hepatitis C was diagnosed in stored sera from 1978 in 24 patients, who were found still to be anti-HCV positive in 1991, and 16 of them were also HCV RNA positive. The majority of anti-HCV positive patients with or without HCV RNA had elevated serum ALT levels 13 years after onset of their acute hepatitis C. After 13 years follow-up, 1.6% of the patients had died of end-stage liver disease, 8% of anti-HCV positive patients had histologically confirmed liver cirrhosis, 79% of anti-HCV positive patients were judged to have chronic infection, whereas 21% seemed to have recovered. To conclude, we found that a majority of our patients with acute symptomatic hepatitis C continued to be viraemic 13 years after onset of hepatitis C, and that all continued to be anti-HCV positive by second-generation ELISA.     

Also with a Restrictive Transfusion Policy,Screening with Second-Generation Anti-Hepatitis C Virus Enzyme-Linked Immunosorbent Assay Would Have Reduced Post-Transfusion Hepatitis C after Open-Heart Surgery

The incidence of post-transfusion hepatitis non-A, non-B (PTH-NANB) was prospectively assessed among open-heart surgery patients from the southeast region of Sweden before the introduction of anti-hepatitis C virus (HCV) blood donor screening. Blood samples for alanine aminotransferase analysis were drawn before and 2, 3, and 4 months after transfusion. Surgery was performed in four centres. Of 190 transfused and followed-up patients 2 (1.1%) contracted PTH-NANB, both operated on at the centre with significantly fewer transfusions than the other centres. One patient had antibodies to HCV detected by first-generation (C100-3) and later by second-generation anti-HCV enzyme-linked immunosorbent assay (ELISA-2) and by positive second-generation recombinant immunoblot assay (4-RIBA). The other patient, although negative by first-generation anti-HCV ELISA, was positive by second-generation ELISA and by 4-RIBA. Both patients were hepatitis C-viremic by polymerase chain reaction (PCR). All the six donors implicated in the two hepatitis cases were first-generation anti-HCV-negative, but two, one for each patient, were positive by second-generation anti-HCV ELISA. This finding was confirmed by positive 4-RIBA in only 1 donor, the other being ‘indeterminate’. However, in both donors hepatitis C viremia was found by PCR. This study shows that the second-generation anti-HCV ELISA will further reduce the risk for PTH-NANB/C and draws attention to the problem of evaluation of confirmatory tests.     

Long-term follow-up of non-A, non-B (type C) post-transfusion hepatitis

One hundred and thirty-five patients who developed non-A, non-B post-transfusion hepatitis mostly after cardiac surgery, were followed for a mean (±S.D.) of 90±41 months (range: 13–180) to evaluate clinical and histological outcome.Thirty-one cases resolved within 12 months, while 104 (77%) progressed to chronicity. Twenty-one of 65 (32%) biopsied patients developed cirrhosis at the end of the follow-up, and one further progressed to hepatocellular carcinoma. One patient had a complete histological remission (1%). The remaining cases had chronic active (37%), chronic persistent (27%) or chronic lobular hepatitis (3%). About half of the cases with cirrhosis developed portal hypertension, and three of these died due to esophageal varices hemorrhage, one due to liver failure, and one due to hepatocellular carcinoma. Out of 26 patients with the initial histologic diagnosis of chronic hepatitis that were rebiopsied during follow-up, 13 (50%) progressed to cirrhosis. These patients were significantly older than patients who did not develop cirrhosis (mean age 57 and 45 years respectively; p<0.01).During acute hepatitis anti-HCV was positive in all but one of the 114 patients tested. Percentages were similar for patients who recovered (95%) and those who developed chronic hepatitis (100%). However, during follow-up, 71% of the 1st generation and 21% of the 2nd generation ELISA test patients with acute resolved hepatitis became anti-HCV negative, while the same figures in chronic cases were only 8.5% (p<0.0001) and 1.4% (p=0.012). This suggests a correlation between anti-HCV antibody activity, hepatitis C virus replication, and the development of chronic liver disease.     

Hepatitis C virus infection in anti-HIV positive and negative French homosexual men with chronic hepatitis: comparison of second- and third-generation anti-HCV testing

ABSTRACT— To determine the prevalence of hepatitis C virus (HCV) infection in homosexuals with chronic hepatitis, we tested for anti-HCV antibodies 113 (47 anti-HIV positive) French non-drug-addicted homosexual men admitted for chronic viral hepatitis. Anti-HCV were detected with second- and third-generation ELISAs (ELISA2 and ELISA3) and RIBAs (RIBA2 and RIBA3). Chronic hepatitis was related to non-A, non-B infection in four, to hepatitis D virus (HDV) infection in five and to hepatitis B virus (HBV) infection in 104 patients. Anti-HCV positivity was found in 50.4% and 12.4% of the 113 patients, with ELISA2 and ELISA3, respectively. Positivity with RIBA2 and RIBA3 was found in only six of the 57 ELISA2 positive patients (all six were ELISA3 positive). The high prevalence of positivity with ELISA2 not confirmed by RIBA2 or RIBA3 suggests false-positive results. ELISA2 positive results were more frequent with frozen serum samples than with fresh serum samples (62% vs 23.5%, p = 0.0003). However, even with fresh serum, ELISA2-positive RIBA-negative results remained frequent in anti-HIV positive patients. ELISA3 seems to give more specific results. We conclude that the prevalence of HCV infection, as assessed with RIBA, was 5.3% among French homosexual men with chronic hepatitis (3.8% after exclusion of transfused patients). This low prevalence suggests that homosexual transmission of HCV is relatively uncommon.     

Prevalence of antibody to hepatitis C virus in prospectively followed acute non-A,non-B hepatitis,from different epidemiological categories

ABSTRACT— We have studied the prevalence of antibody against hepatitis C virus (anti-HCV) and its relation to the time of onset of the symptoms in 57 patients with acute non-A, non-B hepatitis: 16 post-transfusion, 25 drug addicts and 16 sporadic cases. In the 1st month after the onset of illness, anti-HCV was positive in 25% of patients with post-transfusion hepatitis, 44% of drug addicts and 25% of sporadic hepatitis. In the 3rd month this antibody was detected in 75%, 88% and 31.2%, and in the 6th month in 87.5%, 96% and 31.2%, respectively. The prevalence in the 3rd and 6th months was significantly higher in post-transfusion patients and drug addicts than in sporadic cases. In the 6th month the prevalence of anti-HCV in patients who progressed towards chronicity was also significantly higher than in those with acute resolving non-A, non-B hepatitis (94% vs 50%, p<0.001). These results show that HCV is probably the main agent in acute post-tranfusion non-A, non-B hepatitis and in those occurring in drug addicts, and that in a high proportion of these patients the anti-HCV can be detected in the 3rd month after the beginning of the symptoms. On the other hand, the relation of hepatitis C virus with sporadic acute non-A, non-B hepatitis may be doubtful.     

Prevalence of antibodies against hepatitis B virus and hepatitis C virus among blood donors in Lebanon, 1997-2003

The prevalence of hepatitis B virus (HBV) antigens (HBsAg) and antibody to hepatitis C virus (anti-HCV) was determined among 16,084 blood donors (14,993 males; mean age, 31.7 +/- 8.2 years and 1084 females; mean age, 31.4 +/- 8.2 years) in the period 1997-2003. Of the donors screened, 149 were HBsAg positive (0.926%), and 65 were anti-HCV positive (0.404%). There was a steady decline in HBsAg prevalence from 1.56% (1997) to 0.33% (2003) and in anti-HCV from 1.22% (1997) to 0.16% (2003). Females had a higher prevalence of anti-HCV (P = .031) and HBsAg (P = .047). Results obtained are of value in light of the occurrence of HBV and HCV transmission by nonparenteral routes.