电化学法确认低浓度乙肝表面抗原的初步应用

目的利用罗氏试剂公司研发的乙型肝炎病毒表面抗原(HBsAg)确认试剂对低浓度HBsAg标本进行确认试验,并做初步分析,以期对今后的检测工作有一定的指导价值。方法 168例Cutoff-指数(COI)在0.9～20.0血清样本来源于本院住院及门诊患者,经罗氏公司E170仪器对其检测并进行中和试验。结果 94例COI在0.9～5.0低含量HBsAg血清样本,经确认试验确认阳性率72.34%,阴性率26.60%,不确定率1.06%;受试者工作特征曲线(ROC)分析,当COI在4.18时,特异性可达到100%;乙型肝炎病毒表面抗体(抗-HBs)阳性组与阴性组的弱阳性组确认试验情况阳性率为45.71%和88.13%,两组比较,差异均有统计学意义(χ2=19.76,P<0.05)。结论如果条件允许,当HBsAg COI值0.9～5.0IU/L,尤其当同时抗-HBs阳性时应做确认试验。     

电化学法确认低浓度乙肝表面抗原的应用研究

目的 利用罗氏试剂公司研发的乙型肝炎病毒表面抗原（HBsAg）确认试剂对低浓度HBsAg标本进行确认试验,并做初步分析,以期对今后的检测工作有一定的指导价值。方法160例Cutoff指数（C01）在0．9—20．0血清样本来源于本院住院及门诊患者,经罗氏公司e601仪器对其检测并进行中和试验。结果90例COI在0．9～5．0低含量HBsAg血清样本,经确认试验确认阳性率66．7％,阴性率32．2％,不确定率1．1％;受试者工作特征曲线（ROC）分析,当COI在4．2时,特异性可达到100％;乙型肝炎病毒表面抗体（抗一HBs）阳性组与阴性组的弱阳性组确认试验情况阳性率为53．3％和83．3％,两组比较,差异均有统计学意义（P〈0．05）。结论如果条件允许,当HBsAg的COI值0．9～5．0IU／L,尤其当同时抗-HBs阳性时应做确认试验。     

电化学发光法测乙肝表面抗原交叉污染致假阳性灰区

乙肝表面抗原（HBsAg）是乙型肝炎病毒重要的标志物之一，目前多采用ELISA法，灰区的产生多与溶血、HOOK效应、孔的边缘效应等因素有关，电化学发光法具有高灵敏度、宽线性，反应快速、影响因素少、结果准确、自动化程度高等优点而在国内应用越来越广泛，为此，本文对检测HBsAg的交叉污染导致灰区进行了回顾性分析并作一报道：     

时间分辨免疫荧光法和电化学发光法检测HBsAg的对比分析

目的用电化学发光法作为标准,探讨时间分辨免疫荧光法在检测HBsAg上的应用。方法对临床348份血清标本、一份HBsAg高值梯度稀释血清及生物制品检定所标准血清分别用电化学发光法和时间分辨免疫荧光法分别检测,比较两种方法检测结果的阴阳性符合率、灵敏性、特异性、准确性、线性范围以及阳性标本的定量相关性等。结果对临床348份血清检测结果阴阳性符合率为98.26%;阳性标本的定量相关性为γ=0.9641(P<0.001);两种方法的线性范围,时间分辨免疫荧光分析法约为1:2～1:2048(0.21～223.07ng/ml),电化学发光约为1:2～1:4096(0.15～223ng/ml)。结论时间分辨免疫荧光法与电化学发光法在HBsAg的定量检测上的灵敏性、准确性和线性范围都基本相当。     

时间分辨免疫荧光法和电化学发光法检测HBsAg的对比分析

目的 用电化学发光法作为标准,探讨时间分辨免疫荧光法在检测HBsAg上的应用.方法 对临床348份血清标本、一份HBsAg高值梯度稀释血清及生物制品检定所标准血清分别用电化学发光法和时问分辨免疫荧光法分别检测,比较两种方法检测结果的阴阳性符合率、灵敏性、特异性、准确性、线性范围以及阳性标本的定量相关性等.结果 对临床348份血清检测结果阴阳性符合率为98.26%;阳性标本的定量相关性为γ=0.9641(P＜0.001);两种方法的线性范围,时间分辨免疫荧光分析法约为1:2～1:2048 (0.21～223.07ng/ml),电化学发光约为1:2～1:4096(0.15～223ng/ml).结论 时间分辨免疫荧光法与电化学发光法在HBsAg的定量检测上的灵敏性、准确性和线性范围都基本相当.     

电化学发光法和ELISA法检测乙肝血清标志物结果对比分析

目的 探讨电化学发光技术在乙肝血清标志物检测中的应用价值.方法 分别选择132例随机标本和88例乙肝血清标志物ELISA检测少见模式标本,用电化学发光法对其进行检测,并与ELISA结果进行比较.结果 ①在132例随机标本中,ELISA和电化学发光法在对HBsAg,HBsAb,HBeAg,HBeAb和HBcAb检测差异均无统计学显著性意义（P〉0.05）;②在88例临床ELISA检测少见模式标本中,ELISA和电化学发光法在对HBsAg,HBeAg测定中两者差异有统计学显著性意义（χ2=43.00,P〈0.01;χ2=30.03,P〈0.01）;③在88例临床ELISA检测少见模式标本中,电化学发光法检测出其中常见模式有48例（54.55%）.结论 在临床常规标本检测中,电化学发光法与ELISA检测结果相似;但在ELISA检测少见模式标本中,电化学发光法准确性高于ELISA方法.     

ELISA法和电化学发光免疫法检测血清HBsAg结果比较分析

目的对酶联免疫吸附试验（ELISA）和电化学发光免疫法（ECLIA）检测血清中的乙型肝炎病毒表面抗原（HBsAg）结果进行比较分析。方法采用ELISA和ECLIA 2种方法分别检测8 913例血清中的HBsAg。结果 ELISA法和ECLIA法检测出的总阳性数和总阳性率分别为519例占5.82%,451例占5.06%;以ECLIA法为参照,ELISA法假阴性率为0.67%,假阳性率为1.43%,相对敏感性为88.3%,相对特异性为98.5%二者的符合率为97.9%;403例阳性标本用ECLIA法复检,ELISA法的假阳性率为11.9%,相对敏感性为100%,相对特异性为50%,二者的符合率为88.1%;116例弱阳性标本用ECLIA法复检,ELISA法的假阳性率为69%,相对敏感性为100%,相对特异性为50%,二者的符合率为31%;8 394例阴性标本用ECLIA法复检,ELISA法的假阴性率为0.71%,相对敏感性为50%,相对特异性为100%,二者的符合率为99.3%。结论血清HBsAg ELISA法检测存在一定的假阴性率和假阳性率,敏感性和特异性较ECLIA法差,且随HBsAg浓度变化较大。ECLIA法快速、准确、敏感性、特异性更高。     

金标法检测乙肝表面抗原漏检原因分析

目的：分析金标法检测乙肝表面抗原（HBsAg ）的漏检原因。方法采用金标法和酶联免疫吸附法（ELISA）对10335例献血者血液标本进行HBsAg检测,计算阳性率、漏检率。采用金标法和ELISA检测 HBsAg质控品,分析检测灵敏度。结果金标法检测阳性率为0．33％,ELISA为1．14％,金标法检测阳性率低于 ELISA （χ2＝46．76,P＜0．05）。金标法漏检率为71．19％。金标法反应时间为5、10、15 min时,漏检率为94．92％、88．98％、71．19％,比较差异有统计学意义（χ2＝38．27,P＜0．05）。金标法不能检出浓度为0．5 ng/mL的 HBsAg质控品。结论金标法检测灵敏度较ELISA低,并且受人为因素影响较大,适用于献血前初筛。     

分析前因素对血液乙肝表面抗原检测的影响

目的探讨分析前标本处理因素对血液乙肝表面抗原(HBsAg)检测的影响。方法收集300例献血者的血液标本作为研究对象,采用ELISA法进行HBsAg检测。采用配对t检验统计分析对比离心温度为常温(18~27℃)和4℃、采集后储存0 h和8 h离心、离心后2~8℃储存2 h和2~8℃储存2 h后离心、抗凝管和促凝管、常温(18~27℃)和2~8℃储存5种因素对检测结果的影响。结果离心温度、标本采集到离心间隔、储存温度3种因素的t值分别为6.120、-5.694、3.691,对HBsAg检测结果的影响差异有统计学意义(P0.05)。离心温度、标本采集到离心间隔、离心和储存顺序、标本管类型、储存温度5种因素复合标准差分别为0.005 5、0.004 5、0.002 6、0.002 0、0.004 9,影响程度由大到小依次为离心温度、储存温度、标本采集到离心间隔、离心和储存顺序、标本管类型。结论离心温度、标本采集到离心间隔、储存温度3种因素对检测结果有显著影响。环境温度升高和全血标本长时间不分离导致标本不合格,从而对检测结果准确性产生影响,在日常工作中应采取有效措施避免上述因素。     

不同方法检测乙肝表面抗原的结果分析

目的分析酶联免疫吸附法(ELISA)和电化学发光免疫法((ECLIA)检测乙肝表面抗原(HBsAg)的结果及临床应用价值。方法收集经ELISA法检测过的120份临床患者的血清,其中包括:30份HBsAg阳性;30份HBsAg阴性但乙肝e抗体(eAb)和乙肝核心抗体(cAb)阳性;30份HBsAg阴性但cAb阳性;30份乙肝5项均阴性。对上述血清采用ECLIA进行HBsAg检测,并对检测结果进行比较分析。结果 30份经ELISA检测HBsAg阳性者经ECLIA检测均为阳性,其灵敏度达100.0%;30份经ELISA检测HBsAg阴性而eAb、cAb阳性者中,有4份血清经ECLIA检测HBsAg呈阳性,其阳性率达13.3%;30份经ELISA检测HBsAg阴性而cAb阳性者中,有2份血清经ECLIA检测HBsAg呈阳性,其阳性率达6.7%;30份血清经ELISA检测乙肝5项均阴性者中,有2份血清经ECLIA检测HBsAg呈阳性,其阳性率达6.7%。结论 ECLIA法较ELISA法敏感性更高;对临床避免医院内有创检查、手术、输血等情况下的乙肝感染和传播等具有重要应用价值。     

血清中游离乙型肝炎表面抗原的意义

在乙型肝炎患者的血清中存在大量的没有感染性的亚病毒颗粒,这些亚病毒颗粒是由乙型肝炎表面抗原组成的小球形或管型颗粒,其浓度可达到病毒颗粒的100000倍,但对于过量的亚病毒颗粒的产生及其生物学功能目前仍不完全清楚。本文就HBV亚病毒颗粒的生成及其在病毒致病性和临床抗病毒治疗方面的意义进行了阐述。     

Antibody to the hepatitis B surface antigen in immune serum globulin

A collection of 1,278 lots of immune serum globulin (ISG) prepared by 19 United States manufacturers between 1962 and 1974 were tested for the hepatitis B surface antigen (HBSAg) and antibody (anti-HBS). Ten lots (0.8%), all of which were produced between 1962 and 1965 by two different manufacturers, were weakly positive for HBSAg (by radioimmunoassay). Seven hundred and seven lots (55.3%) were positive for anti-HBS (by passive hemagglutination). In general, titers of anti- HBS in lots of ISG were low, and the prevalence of anti HBS positive lots varied considerably among different manufacturers. ISG prepared from placental material was more commonly positive for anti-HBS than was ISG prepared from plasma. There was a striking overall increase in prevalence and titer of anti-HBS in ISG lots prepared during 1973 and 1974. This probably reflects the effect of elimination of strongly HBSAg-positive plasma units with the onset of routine screening for HBSAg which began in 1972.     

Chronic hepatitis B and hepatitis B surface antigen carriers in university students

A study was made of latent hepatic diseases in university students. 1.4% to 1.6% of the students were positive for hepatitis B surface antigen (HBsAg), and 10.3% for anti-HBs. Of 28 students with HBs-antigenemia, 2 had chronic persistent hepatitis, and 3 minimal hepatitis, 23 being healthy carriers. Hepatitis B e-antigen (HBeAg) was detected in 44% of the students with HBsAg, and anti-HBe in 13%. Anti-HBe was significantly more frequently found in female students with HBsAg than in male students. Though most of the students with HBsAg had high titer of antibody to hepatitis B core antigen (anti-HBc), there were a small number of cases showing low titer. HBsAg and anti-HBs was detected in the same serum specimens of 2 carrier students. Liver damage was also found in 3 students without HBs-antigenemia.     

Hepatitis B virus DNA and hepatitis B surface antigen levels in chronic hepatitis B

Despite universal vaccination, chronic hepatitis B (CHB) continues to be a major health burden worldwide, with an estimated 350–400 million people infected with the virus. Over the past decade, rapid progress has been made with regards to antiviral therapy for CHB, from conventional interferon to pegylated interferon, and with the earliest oral agent lamivudine to the current, more potent drugs such as entecavir and tenofovir. There have also been new developments in the diagnostic and monitoring tools for CHB. Qualitative hepatitis B surface antigen (HBsAg) testing has been used to diagnose patients infected with CHB. More recently, quantitative HBsAg titers have been used to predict treatment outcome when measured at baseline or early into treatment. The progress on the use of hepatitis B virus (HBV) DNA levels has been more rapid. Serum HBV DNA levels have been shown to be important in the natural history of CHB infection, with higher levels being significantly associated with the development of cirrhosis and hepatocellular carcinoma. For patients receiving antiviral therapy, the baseline and early on-treatment HBV DNA levels are important in determining treatment outcomes. Monitoring of HBV DNA levels during therapy will allow for early detection of drug resistance. The end-of-treatment and post-treatment HBV DNA levels have been demonstrated to be important indicators of treatment success and relapse, respectively. With newer and more powerful antiviral agents, and with the development of quantitative assays that are highly sensitive, further studies are needed to optimize the use of these tools and agents in the modern management of CHB.     

FibroTouch检测肝脏硬度评价慢性乙型肝炎肝纤维化程度的影响因素分析

目的：探讨影响FibroTouch检测肝脏硬度(LSM值)评价慢性乙型肝炎肝纤维化程度的相关因素。方法：收集2015年1月至11月就诊经肝组织穿刺活检确诊的慢性乙型肝炎病毒感染者144例,应用FibroTouch技术检测肝脏LSM值,同时收集患者一般临床信息及常规血液检测指标。将筛选出的自变量进行多元线性回归分析,建立回归方程,并对回归模型进行评估。结果：经多元线性回归筛选出6个独立预测因子,分别为：脾脏肋间厚度(脾厚),体质指数(BMI),血清白蛋白(ALB)、总胆红素(TBiL)、凝血酶原时间(PT)、IV型胶原(CIV),以此建立回归方程。该回归模型决定系数R方=0.648,调整R方=0.609,具有良好的拟合度。结论：在进行FibroTouch 检测LSM时,脾厚、CIV、ALB、TBiL 、PT和BMI值,可能会对检测结果产生影响,对于影响因素的探讨仍有待进一步的大样本多中心研究。     

乙型肝炎前S1抗原和乙型肝炎血清标志物联合检测结果分析

目的分析乙型肝炎病毒前S1抗原与乙型肝炎病毒(HBV)血清标志物的相互关系。方法用酶联免疫吸附试验对门诊、住院患者血清进行乙型肝炎5项,前S1抗原联合检测(将检测结果中除抗-HBs阳性外的其余项中至少有一项阳性的1591例患者检测结果作回顾性分析)。结果 1591例标本检测结果中,前S1抗原主要出现在HBsAg阳性血清中,前S1抗原在HBeAg阳性组合中的检出率显著高于HBeAg阴性组合,差异有统计学意义(P0.05),在"大三阳"组合中的检出率显著高于"小三阳"组合,差异有统计学意义(P0.05),前S1抗原、HBeAg检出率差异有统计学意义(P0.05)。结论前S1抗原与其他乙型肝炎表面标志物有一定的相关性,在实际工作中两种方法可协同检测,相互补充。     

Structural analysis of hepatitis B surface antigen by monoclonal antibodies.

A method has been developed for the analysis of hepatitis B surface antigen (HBsAg) antigenic structure at the molecular level that creates "fingerprints" or "signatures" of various hepatitis B viral (HBV) strains. This technique employs high affinity IgM and IgG monoclonal antibodies (anti-HBs) directed against distinct and separate determinants on HBsAg. In performing this antigenic structural analysis, separate binding curves for different monoclonal anti-HBs are generated by measuring immunoreactivity in serial dilutions of HBsAg-positive serum by radioimmunoassay. Since the HBsAg concentration in serum is unknown, the binding profiles of groups of samples are aligned by an iterative least-squares procedure to generate the numerical signature characteristic of the viral strain. The numerical signatures are then displayed on a computer-graphic plot. The signature profiles of HBsAg subtypes are a true reflection of their antigenic structure, and in vertical and horizontal transmission studies the molecular characteristics of the viral epitopes are conserved. By signature analysis we found substantial antigenic heterogeneity among the ayw3 strain both in the U.S. and France, as well as in populations of the Far East and Africa. Populations in Ethiopia, Gambia, and the Philippines were infected with two antigenically distinct HBV strains. In some newly identified HBV strains, it was found that epitopes identified by some monoclonal antibodies were absent or substantially reduced, which suggested that a genetic mutation may have occurred. Thus this study suggests that there is far more antigenic heterogeneity in HBV than previously recognized. These variants are antigenically distinct from each other at the epitope level, and were heretofore unrecognized by polyvalent anti-HBsAg antibodies.     

Antibody to hepatitis B surface antigen: immunochemical purification from standard immune serum globulin

Hepatitis B surface antigen (HBsAg) may be insolubilized onto a controlled pore glass (CPG) matrix by simple adsorption from acidified serum or plasma. The resultant complex has the properties of an HBsAg- specific immunoadsorbent. In this study, CPG-HBsAg complexes have been used to adsorb anti-HBs from standard immune serum globulin. Specific antibodies were eluted with acidic or chaotropic buffers. Recoveries ranging from 29 to 46 per cent, with purifications of approximately 1000-fold were obtained. Two procedures designed to inactivate hepatitis B virus had no effect upon the immunoadsorbent properties of the CPG-HBsAg complex. The possibility of using this simple method to prepare hepatitis B immune globulin is discussed.