国产化学发光免疫试剂检测血清HBsAg的应用评价

目的考核国产化学发光免疫分析试剂检测血清HBsAg的效果，探讨其在血液筛查和临床检测应用中的可行性。方法采用国产化学发光免疫分析试剂筛查345份HBsAg确认阳性的血清标本，比较其与常用的进口和国产酶联免疫检测试剂盒的漏检率；并采用不同浓度的标准HBsAg血清对国产化学发光免疫分析试剂进行线性分析和精密性试验。结果国产化学发光免疫分析检测试剂的漏检率高于Murex和Organon酶联免疫试剂盒，差异有统计学意义（P〈0．01），而与国产英科新创（厦门）科技公司和上海科华生物技术有限公司的HBsAg酶联免疫诊断试剂盒的差异无统计学意义；采用标准血清进行线性分析，标本浓度在（0．5～150）ng／ml范围时与化学发光单位量（RLUs）呈良好的线性正相关；分别对低、中、高浓度标准血清进行板内和板间的重复检测，其反应具有良好的重复稳定性。结论化学发光免疫分析法检测血清HBsAg具有快速、可定量、反应稳定的特点，可用于血液筛查和临床标本的检测。     

HBsAg常规检测阴性结果分析

目的 了解日常工作中HBsAg漏检情况,为提高临床检验质量提供参考.方法 在日常工作中从临床检测备份管随机收集400份经国产A品牌全套试剂ELISA检测乙型肝炎病毒血清学指标(HBVM),其中HBsAg呈阴性结果的血清标本,按HBsAb检测结果分成HBsAb阴性和阳性两类各200例,双份分装-20℃冻存;增加B,C另外两种国产ELISA HBsAg试剂及美国超敏MONOLISA HBsAg ULTRA试剂,共四种试剂同时复检HBsAg,阳性标本行双份HBV DNA定量分析、结果取均值.结果 A,B,C三种国产试剂复检HBsAg结果一致阴性,美国超敏MONOLISA HBsAg ULTRA试剂从HBsAb阴性标本中检出5例HBsAg阳性,从HBsAb阳性标本中未检出HBsAg阳性结果.HBV DNA定量分析结果显示均小于500 copies/ml,其中400～500 copies/ml 1例, 100～200 copies/ml 2例,10～100 copies/ml 2例,未发现低于检测下限结果.结论 国产常规ELISA HBsAg试剂灵敏度普遍偏低、容易漏检;漏检标本多来自HBsAb阴性个体;漏检个体可能和隐匿性HBV感染相关,临床应重视长时间HBsAb阴性个体的HBVM定期复查工作.     

金标法与ELISA法检测HBsAg的比较

目的对无偿献血者标本采用金标法快速检测HBsAg,并与ELISA法检测结果进行比较,分析金标法漏检原因,寻找改进措施。方法对金标法检测HBsAg后血液标本留样,采用2种不同ELISA试剂进行2次HBsAg检测。结果 31146份无偿献血者标本中,用金标法检出阳性69份,阳性率0.22%;ELISA法检出阳性286份,阳性率0.92%,金标法和ELISA法检测结果的差异有统计学意义（χ^2=7031,P〈0.01）。对ELISA法检出阳性的286人份用金标法复查,在试纸反应5、10、15min后的漏检率分别为91.3%、77.2%、60.3%。结论金标法检测HBsAg漏检率较高。应将其与ELISA检测相结合。     

7家ELISA法HBsAg诊断试剂的质量评价分析

目的对7家ELISA法HBsAg诊断试剂(国产5家进口2家)进行质量考核和分析,为实验室选择高质量试剂提供参考。方法应用临床科研血清盘、HBsAg弱阳性定值质控血清,对7家ELISA法HBsAg诊断试剂盒的灵敏度阴阳样总符合率以及亚型最低检出限进行质量评价分析。结果 7家试剂对HBsAg弱阳性0.2IU/mL定值质控血清、adr、adw亚型的最低检测限,检测的S/CO比值的差异很大;3国产试剂对ay血清型的出现漏检现象。结论 7家HBsAg ELISA诊断试剂的灵敏度总符合率均有较大的差异,实验室在选择试剂时应当进行考核,进口试剂相对质量较好。     

ELISA法HBsAg检测试剂盒质量评价

目的比较国产ELISA法HBsAg检测试剂盒的质量差异,优选适宜于献血者HBsAg检测的试剂盒。方法应用HBsAg国家参考品对所购进的试剂盒进行质量评价。结果5个厂家试剂的阴性符合率、阳性符合率、总符合率、灵敏度和精密度存有一定差别,2个厂家试剂有假阳性出现,2个厂家的试剂有最低检出限量参考品未检出。结论不同厂家的试剂质量存在明显差异,因此试剂使用前必须进行质量抽检,综合评价其质量。选择灵敏度高、特异性强的试剂,以提高献血者HBsAg的检测质量。     

ELISA法检测HBsAg灰区和单试剂反应性样本的确认结果分析

目的 探讨ELISA法检测HBsAg结果判定灰区设置的必要性,以及中和试验对HBsAg灰区和单试剂反应性样本的确认情况.方法 采用两个不同厂家生产的ELISA HBsAg试剂对献血者标本分别进行检测,对检测结果在0.7≤S/CO＜1或单试剂为反应性的标本,再用同种试剂进行双孔复试,双孔中至少有1孔结果仍在灰区内或仍为反应性,留取标本做HBsAg中和试验确证.结果 136例灰区标本中HBsAg中和试验检测出阳性标本10例,阳性率为7.35％,结果异常标本14例;159例单试剂反应性标本中HBsAg中和试验检测出阳性标本19例,阳性率为11.95％,结果异常标本16例.灰区与单试剂反应性样本阳性率的差异没有统计学意义;S/CO值在0.70～0.79区间、0.80～0.89区间、0.90～0.99区间的阳性率有逐渐升高的趋势,但差异无显著性;双试剂灰区的阳性率高于单试剂灰区的阳性率,但差异无统计学意义.结论 对HBsAg灰区和单试剂反应性标本,ELISA检测漏检率较高,应结合自己实验室实际设置合适的灰区,最大限度地检出这些可疑标本,并进行确认实验,进一步减少HBsAg漏检和误检率,以提高输血安全,同时避免血源的浪费.     

快速金标法检测HBsAg漏检原因的分析

随着《献血法》的实施,无偿献血工作已在我国全面展开。为便于公民无偿献血,本中心从1999年5月起采用快速HBsAg全血金标试条〈简称金标法〉对献血者进行HBsAg初筛,发现存在一定的漏检。为了解漏检原因,对金标法初筛合格的18123份血液,经ELISA法复检,根据ELISA法检测结果,以及追踪观察保存的现场初筛HBsAg金标试条等结果,对漏检原因进行了分析,报告如下。 1 材料与方法 1.1 试剂 HBsAg-ELISA试剂盒,批号：9030501、9960501,灵敏度：0.5ng/ml、0.2ng/ml;HBsAg金标试剂,批号：9051901、9081201,灵敏度：1ng/ml,均为厦门新创有限公司产品。 1.2 标本来源 1999年5月～2000年2月本中心现场无偿献血,金标法初筛合格采集的18123份血液标本。 1.3 方法对现场金标法初筛合格后采集的血液用ELISA法常规检测,对检出的阳性及可疑标本,再用该法双孔复检,其中任一孔S/CO值≥1确定为阳性;同时再次用金标法作对应检测,并追踪观察保存的现场初筛HBsAg金标试条。     

金标法快速检测无偿献血者HBsAg漏检原因分析

目的对无偿献血者金标法快速初筛检测HBsAg漏检原因进行分析,寻找改进措施。方法金标法初筛HBsAg阴性后采集的血液,留取标本用两种酶标ELISA试剂及不同人员进行两次复检,确定金标法初筛HBsAg检测情况。结果对47507人次金标法初筛HBsAg阴性献血后标本用酶标ELISA法复检有513份标本为阳性,阳性率1.1%,再用同一厂家的金标试剂复测,出现六种不同结果。结论灵敏度和批间差是金标漏检的主要原因,加强操作人员的技能培训和工作责任心,改善工作环境能有效减少HBsAg漏检。     

金标法快速检测无偿献血者HBsAg漏检原因分析

目的 对无偿献血者金标法快速初筛检测HBsAg漏检原因进行分析,寻找改进措施.方法 金标法初筛HBsAg阴性后采集的血液,留取标本用两种酶标ELISA试剂及不同人员进行两次复检,确定金标法初筛HBsAg检测情况.结果 对47507人次金标法初筛HBsAg阴性献血后标本用酶标ELISA法复检有513份标本为阳性,阳性率1.1%,再用同一厂家的金标试剂复测,出现六种不同结果.结论 灵敏度和批间差是金标漏检的主要原因,加强操作人员的技能培训和工作责任心,改善工作环境能有效减少HBsAg漏检.     

G145R变异重组HBsAg ELISA检测质控参照品制备的初步研究

目的：实验性制备G145R变异重组乙型肝炎表面抗原（rHBsAg）质控参照品。方法：收集合G145R变异rHBsAg的细胞培养上清，采用50％饱和硫酸铵盐析．根据其在D12-ELISA中的检测信号，以卫生部颁布的HBsAg ELISA质控血清精确标定盐析物中靶物质的含量，适量稀释分装。置-20℃冻存。采用不同方法对制备物的特异性与稳定性进行考核．并将其试用于对几种市售试剂盒检测变异能力的评价。结果：制备物中靶物质含量在0．50～32.00ng／mL wcHBsAg之间，在D12-ELISA检测中其信号的线性关系与质控血清有很好的一致性；反复冻融对其稳定性无显著影响：采用不同市售ELISA试剂检测时其反应信号强度各不相同。结论：本研究制备的G145R变异rHBsAg ELISA质控参照品具有较高的稳定性、特异性与实用性；其研制为乙肝病毒免疫逃逸现象在基础、临床、诊断试剂研发，以及流行病学调查等方面研究的开展提供了重要的物质基础。     

A new HBsAg screening assay designed for sensitive detection of HBsAg subtypes and variants

The design of a new HBsAg screening assay, the Hepanostika HBsAg Ultra is based on the use of monoclonal antibodies raised against native wild-type HBsAg and reactive with HBsAg in which the common 'a'-determinant is modified by site-directed mutagenesis of four of the cysteine moieties. The design was checked using the same cysteine variants and samples from patients known to be infected with HBsAg variants. The results found were compared with other state-of-the-art commercial screening assays. The design of the Hepanostika HBsAg Ultra enabled detection of all variant HBsAg-positive samples in contrast to the other commercial assays. An additional 980 samples were tested to assess the specificity and sensitivity of the Hepanostika HBsAg Ultra. Screening of presumed negative serum and plasma samples resulted in a specificity of 100%. This makes the Hepanostika HBsAg Ultra the first screening assay with a design able to detect HBsAg variants with high sensitivity and specificity.     

Development of an economic and efficient strategy to detect HBsAg: application of "gray-zones" in ELISA and combined use of several detection assays

BackgroundELISA and CMIA are commonly used for detection of HBsAg. However, few investigations have been performed to evaluate their value in clinical practice, especially when jointly used. A reasonable and economic HBsAg testing algorithm is in great need.MethodsA total of 161,426 specimens in China were tested for 5 serum HBV markers with commonly used ELISA kits. 498 of these specimens were further tested for HBsAg by another ELISA kit, a CMIA kit and an HBsAg confirmatory assay.ResultsThe sensitivities of the 2 ELISA kits were 76.21% and 88.42%, respectively. However, when using “gray-zones”, the sensitivities were significantly improved to 97.43% and 96.43%. Furthermore, the combined use of the 2 ELISA kits and their “gray-zones” improved the sensitivity to 99.04%. Nevertheless, 2.91% of the samples with S/CO values below the lower “gray-zone” limits were reactive by the CMIA kit and then confirmed as HBsAg positive. However, 71.43% of the samples with HBsAg values within 0.05 and 0.10 IU/ml detected by the CMIA kit could not be confirmed.ConclusionsAs a rational and economic strategy, combined use of “gray-zones” in ELISA and several different detection assays can significantly increase the efficiency of HBsAg detection.     

Evaluation of sensitivity for wild type and mutant forms of hepatitis B surface antigen by four commercial HBsAg assays

Hepatitis B surface antigen is one of the most important serological markers used to diagnose an HBV infection. Improvements in HBsAg assay sensitivity have been achieved constantly over the years since introduction of the first commercial assays. It is generally assumed that diagnostic assay sensitivity includes the ability to detect wild type and viral variants at the same level. Thus it would be expected, as newer HBsAg assays are developed, that viral mutation would be considered in assay design and that assay sensitivity would be equivalent for wild type as well as mutant forms. Two newly launched HBsAg assays (Bayer ADVIA Centaur and Ortho VITROS ECi) were compared to two established HBsAg assays (Abbott AxSYM and Roche Elecsys) in order to test the assumption that assay sensitivity for variants is equivalent to wild type HBsAg. The four assays were challenged with a standard HBsAg sensitivity panel of both ad and ay subtypes as well as a 13 member mutant panel comprised of both recombinant and native HBsAg members. Results demonstrate that the analytical sensitivity for wild type HBsAg is comparable for all assays tested. In contrast, significant differences were observed for detection of mutants. AxSYM HBsAg detected all mutant samples while all other asssays missed 10 out of 13 samples tested. It is noteworthy that the most frequently reported HBsAg mutation, G145 R, remained undetected in three of the assays tested. It is discussed whether the reduced sensitivity for mutants of the most recent assays represents a new risk for the diagnosis of HBV infection.     

Comparative evaluation of tree commercial diagnostic kits for detection of M. tuberculosis by polymerase chain reaction

Three Russian commercial PCR diagnostic kits for detection of M. tuberculosis in clinical samples are compared. The kits were evaluated by sensitivity and convenience of analysis. The specificity and sensitivity of Russian diagnostic kits are not inferior to foreign analogs, but are not devoid of some drawbacks.     

应用CLSI EP12-A2文件评价HBsAg酶联免疫吸附试验试剂盒的分析性能

目的：应用CLSI EP12-A2文件评价乙型肝炎病毒表面抗原（HBsAg）的酶联免疫吸附试验（ELISA）试剂盒的性能,并比较2个不同厂家试剂盒之间的性能差异。方法按照CLSI发布的EP12-A2文件,对实验室使用的2个不同厂家的 HBsAg ELISA试剂盒的性能进行评价。结果2个厂家的试剂盒对分析物浓度为120％临界值的标本的检测阳性率均大于或等于95％,对分析物浓度为80％临界值浓度的标本的检测阴性率大于或等于95％,批内变异系数小于或等于15％,批间变异系数小于或等于20％。2个厂家试剂盒的灵敏度分别为94．2％（95％CI：84．3％∽98．0％）和92．3％（95％CI：81．8％∽97．0％）;特异性分别为98．0％（95％CI：89．5％∽99．6％）和100．0％（95％CI：92．8％∽100．0％）。2个厂家试剂盒灵敏度差值95％区间为-5．46％∽10．19％,特异性差值95％区间为-2．00％∽5．32％。结论2个厂家试剂盒检测 HBsAg 临界值＆#177;20％的浓度范围之外标本,能够得到稳定、可靠的检测结果,并具有较高的灵敏度和特异性,两者的检测灵敏度和特异性相近。     

Variation in the sensitivity of HBsAg screening kits

Summary. Fifteen HBsAg kits from 14 manufacturers were assessed. Their sensitivity was evaluated by testing 150 HBsAg-positive sera, sera from four donors who were low-level HBsAg carriers, and sequential specimens from 22 seroconverting individuals together with dilutions of six of these specimens. The British HBsAg Working Standard (0.5IUmL^-1) and the NIBSC/UKBTS HBsAg Monitor Sample (0.125IUmL^-1) were also tested. Five assays failed to detect one of the 150 routine HBsAg-positive sera. Four assays (Auszyme Monoclonal; Monolisa Ag HBs 2nd generation; Murex HBsAg; Ortho HBsAg Test Systems 3) were able to detect HBsAg in all but one of the six sera from low-level carriers, whereas one assay (MicroTrak II HBsAg) detected only one of the six. The most sensitive kit (Monolisa Ag HBs 2nd generation) detected HBsAg in 79 specimens from the seroconversion panels; four other kits detected HBsAg in at least 70 specimens, seven in 60–69, two in 50–59 and the least sensitive in 31. Further analysis of the findings on seroconverters indicated a median reduction in the duration of HBsAg detection of 5 days or more for four assays when compared with the most sensitive assay. One kit (Auszyme Monoclonal) detected HBsAg in 15 of the 18 dilutions prepared from the seroconversion specimens, whereas three kits detected HBsAg in fewer than 10 dilutions. Two kits gave negative reactions with the British HBsAg Working Standard on all of five occasions and six were consistently unreactive with the NIBSC/UKBTS HBsAg Monitor Sample; only three kits (Bioelisa, Enzygnost, Murex) were always reactive. There is therefore substantial variation in sensitivity among the HBsAg kits currently available.     

强光板消毒仪用于口腔科印模托盘正畸模型消毒的研究

目的 为口腔科印模托盘、正畸模型寻找一种安全可靠的消毒方法 ,弥补传统消毒方法 存在的不足.方法 采用自行研制的强光板紫外线消毒仪,经染菌实验、染HBsAg破坏试验及监测观察组与对照组122件印模托盘正畸模型消毒前后含菌量对比,观察消毒效果.结果消毒5 min染菌杀灭率为99.99%,符合卫生部对指示菌杀灭率≥99.9%的标准要求;消毒18 min时,HBsAg抗原性破坏符合卫生部S/N<2.1标准,20 min其抗原性完全破坏;监测122件印模托盘正畸模型物品,其消毒前后含菌量有显著差异. 结论 采用强光板紫外线消毒仪消毒物品,具有对人体无危害,对环境无污染,消毒时间短的优点,为口腔科印模托盘、正畸模型的消毒提供了一种安全、可靠、易操作的方法 .     

电化学发光免疫法和化学发光微粒免疫分析法在HBsAg检验中的价值比较

目的比较电化学发光免疫法(CI)和化学发光微粒免疫分析法(CMIA)在乙肝表面抗原(HBsAg)检验中的价值。方法纳入我院收治的乙型肝炎疑似病例396例,分别使用CMIA、CI法进行HBsAg水平的测定。以CMIA为金标准,分析CI对HBsAg的诊断灵敏度、特异度和符合率。计算CMIA、CI检测HBsAg的检测批次内和批次间精密度。使用Bland-Altman偏差分析对CI、CMIA的诊断一致性进行分析。结果以CMIA为检测金标准,CI检测的诊断灵敏度为95.03%(287/302),诊断特异度为97.87%(92/94),诊断符合率为95.71%(379/396)。CI、CMIA对HBsAg检测的批次内变异系数和批次间变异系数比较,差异无统计学意义(P>0.05)。CI和CMIA对HBsAg诊断的组内相关系数为0.896,CI、CMIA在对HBsAg的诊断中,一致性较好。经Bland-Altman偏差分析得,两种检测方法的偏倚程度置信区间为(-6.36~23.24),CI、CMIA在0~23 IU/mL的区域内具有良好的一致性,随着机体HBsAg水平的增加,偏差逐渐上升,2.02%(8/396)的点位于偏倚程度95%置信区间以外。结论与CMIA相比,CI在对HBsAg的检验中具有良好的诊断灵敏度、符合率以及精密度。此外,CMIA、CI对HBsAg的诊断一致性良好,然而在高水平HBsAg的检测中仍然存在一定偏差。     

Cross-reactivity of monomeric and dimeric biosynthetic human growth hormone in commercial immunoassays

Commercial kits give different measurements for concentrations of growth hormone (GH, somatotropin) in serum. Most notably, a two-site monoclonal-antibody-based immunoradiometric assay (IRMA) from Hybritech routinely yields lower values than do conventional RIAs in which polyclonal antibodies are used. We used purified dimeric biosynthetic human GH as a model compound to investigate the specificity of five commercial immunoassays for size variants of GH. In all five assays, biosynthetic monomeric GH was significantly more potent than pituitary-derived standard GH supplied with the kits. Dimeric GH was significantly less potent than monomer in four of the five assays, and cross-reactivities varied more than fivefold, from 15% to 84%. Using three commercial kits selected for their specificity for dimeric GH, we measured GH in serum samples from 18 normal adults. The mean GH concentrations in serum--0.7 (Hybritech, IRMA), 1.8 (Diagnostic Products, RIA), and 3.1 (Cambridge, RIA) micrograms/L--differed significantly, but in the same rank order as that obtained in the experiments on dimer cross-reactivity.