酶校正品在血清酶测定中的应用

目的　应用酶校正品校准不同的“仪器/试剂”测定系统,观察系统间血清酶测定结果的可比性。方法 　以“日立仪器/cfas/罗氏试剂”为参照系统,用cfas对各系统作校正,测定科华及中生试剂在校正前、后的比例误 差与恒定误差。结果　科华丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、γ 谷氨酰基转移酶(GGT)的 比例误差由校正前-17.7%、8.5%、-26.1%变为5.2%、4.1%、0%;恒定误差则由校正前-1.494U/L、-7.290 U/L、2.620U/L变为-2.326U/L、-4.461U/L、4.856U/L;中生ALT、AST、GGT的比例误差由校正前-31.4%、 -23.9%、-27.8%变为-9.3%、-13.4%、-6.7%;恒定误差则由校正前0.0001U/L、-0.451U/L、0.035U/L 变为0.956U/L、3.727U/L、2.236U/L。结论　各系统经校正后的测定结果更接近于参照系统测定值但仍存在一 定的系统误差,需要将系统间存在的比例误差及恒定误差进行校正才能与参照系统的结果相一致。     

酶校准品在血清酶测定结果一致性中的应用

目的探讨在不同生化分析仪上检测常用血清酶[丙氨酸转氨酶(ALT)、门冬氨酸转氨酶(AST)、γ-谷氨酰转移酶(GGT)、碱性磷酸酶(ALP)、乳酸脱氢酶(LDH)、肌酸激酶(CK)]结果的可比性的方法。方法将自动化系统的校准物(cfas)中6个酶活性定值转移至新鲜混合血清,以该混合血清作为校准品,对同一批标本测定校准前后的6个酶活性。结果校准前,同一批标本在2台不同品牌仪器上测得的结果差异有极其显著性(P<0.001)。校准后,同一批标本在2台不同品牌仪器上测得的结果差异无显著性(P>0.05)。结论用cfas转移血清酶定值的新鲜混合血清作为校准品对不同检测系统进行校准,既保证了结果的可溯源性,又可以实现血清酶测定的一致性。     

压电免疫传感器用于抗丙型肝炎病毒抗体检测的实验研究

目的 研制一种用于检测抗丙型肝炎病毒(HCV)抗体的压电免疫传感器.方法 采用聚乙烯亚胺黏附、戊二醛交联的方法,选用(HCV)人工合成多肽抗原作为生物敏感材料,研制成一种新型的抗HCV抗体压电免疫传感器,将其用于临床血样的检测,探索了传感器的组装条件,考察了传感器的响应特性.结果 HBsAg对抗HCV抗体检测干扰较小,且抗HCV抗体阳性血清的响应值与阴性血清的噪声值之比大于4.结论 该方法选择性好,易于操作,能快速有效地检测抗HCV抗体.     

血清酶测定标准化的实验研究

目的　通过酶校 (标 )准品在临床实验室中的应用 ,探讨血清酶测定标准化的新途径。方法　向参加实验室 (15 0家 )发放酶校准品 1支和血清样本 3支 ,各实验室用现行方法分别测定校准品和血清样本 ,记录检验结果 ;用校准品定标后再测定血清样本 1次 ,记录检验结果 ;计算校准品测定结果总体均值与标示值间的偏倚和校准前后血清样本测定结果精密度的变化。结果　丙氨酸氨基转移酶 (ALT)、天门冬氨酸氨基转移酶 (AST)、淀粉酶 (AMY)和乳酸脱氢酶 (LD)的测定结果均值与标准品标示值的相对偏倚≤± 6 % ,分别为 3 8% ,- 1 8% ,2 3%和 - 5 2 % ;碱性磷酸酶 (ALP)、γ 谷氨酰基转移酶 (GGT)和肌酶激酶 (CK)的实验室检测结果与标准品标示值间的相对偏倚小于 2 0 % ;分别为13 7% ,- 13 9%和 - 19 2 %。在应用校准品后 ,多数项目测定结果的变异系数 (CV % )得到不同程度的改善 ,ALT由 10 %～ 4 4 %下降至 8%～ 4 2 % ,LD由 16 %～ 39%下降至 8%～ 11%。结论　我国临床实验室对血清ALT、AST、AMY和LD的测定结果有较好的向国际标准溯源性 ;使用酶校准品可改善不同分析系统间测定结果的一致性。     

艾滋病病毒抗原检测试剂盒研制及应用

目的建立双单抗夹心酶联免疫法检测人类免疫缺陷病毒核心抗原（ＨＩＶ－１ｐ２４）的试剂盒。方法用识别ＨＩＶ－１ｐ２４抗原不同位点的单克隆抗体，构建双单抗夹心酶联免疫法，检测ＨＩＶ－１ｐ２４抗原，测定试剂盒的临界值及灵敏度；对４５２份抗ＨＩＶ阳性样品及１５２５份高危人群抗ＨＩＶ阴性样品进行测定，用抗体阻断抑制法确证，并与聚合酶链反应病毒核酸测定结果进行比较。结果获得６株单抗，免疫印迹法测定结果显示均为ｐ２４特异性。交叉组合试验确定单抗ｐ２４－１和１０用于固相，ｐ２４－６用于标记辣根过氧化物酶。试剂盒临界值为０．１５＋阴性对照值；检测灵敏度为２ｎｇ／ｍｌ。４５２份抗体阳性样品，抗原检测阳性４９份（１０．８％），１５２５份抗体阴性样品，经阻断试验确证，抗原阳性１７份。４９份抗体阳性样品中，ＨＩＶ－ＲＮＡ阳性３６份，符合率达７３％；１７份高危人群抗原阳性样品，１０份为病毒核酸阳性。结论双单抗夹心酶联免疫检测ＨＩＶ－１ｐ２４抗原方法敏感、操作简便，可用于临床ＨＩＶ感染的辅助检测手段。     

CEDIA法测定血清地高辛的实验观察与临床应用

目的克隆酶供体免疫分析技术（CEDIA）与放免法（RIA）测定血清地高辛的两种实验方法的观察比较。方法采用CEDIA对206例心脏疾病患者的血清地高辛浓度进行测定。结果经统计学分析，151例非中毒组和55例中毒组血清地高辛浓度分别为1．23±0．46ng／ml和2．59±0．62ng／ml。结论实验结果与国外报道接近，与放射免疫分析进行相关实验，结果r=0．9263，相关良好。     

血清粗纤维调节素ELISA的建立及初步应用

以提纯的人胎盘粗纤维调节素 (Un)及兔抗人Un IgG ,建立血清Un的竞争抑制ELISA标准曲线 ,并检测了 6 0名正常人和 12 4例肝病患者。结果采用棋盘试验确定包被Un及其抗体的浓度 ,表明该法灵敏度为 5 μg/L ,回收率为 96 6 %～10 8 3% ,批内 CV为 7 5 % ,批间CV 为 7 9%。 6 0名正常成人血清Un含量为 71 14± 2 9 6 0 μg/L ,参考值范围为 11～ 130μg/L,慢性肝病患者Un含量显著高于正常人。提示此法稳定、可靠、特异 ,基本符合临床使用要求 ,可用于慢性肝病的诊断。     

多通道电化学传感器检测人血清总前列腺特异性抗原

目的建立一种简便、灵敏和准确的多通道电化学传感器分析方法(ECISA),用于人血清总前列腺特异性抗原(tPSA)的检测,并进行性能评价。方法基于抗体真空组装技术,将抗PSA抗体在16通道碳印刷电极阵列表面进行自组装,结合双抗体夹心免疫分析原理和多通道电化学传感技术对185份人血清中的tPSA进行检测,并将结果与电化学发光免疫分析法(ECLIA)进行相关性分析。结果多通道电化学传感技术检测tPSA标准曲线的线性范围为0.05~50 ng/mL;tPSA的最低检测限为10 pg/mL;定量限为61 pg/mL;检测tPSA批内变异系数(CV)为8.00%~8.50%;与ECLIA定量结果相关性良好(r2=0.941,P0.01)。结论 ECISA技术检测tPSA的灵敏度和特异性均较高,且与ECLIA结果有较好的相关性,能满足临床对tPSA检测的要求,具有潜在的临床推广应用价值,并具备用于其他肿瘤标志物检测的前景。     

A disposable multianalyte electrochemical immunosensor array for automated simultaneous determination of tumor markers

BACKGROUND: Automated and convenient multianalyte detection with high throughput is increasingly needed in clinical diagnosis. We developed a disposable 4-by-2 array for programmed simultaneous amperometric immunoassay of 4 tumor markers. METHODS: We used a screen-printed technique, 1-step immobilization method, and flow injection technique. We immobilized carcinoembryonic antigen, alpha-fetoprotein, beta-human choriogonadotropin, and carcinoma antigen 125 as model analytes in a redox mediator-grafted, biopolymer-modified, screen-printed carbon electrode array to capture corresponding horseradish peroxidase-labeled antibodies in competitive immunoreactions. The simultaneous multianalyte immunoassay was automatically carried out to amperometrically monitor the mediator-catalyzed enzymatic response to hydrogen peroxide, which decreased in proportion to the concentrations of analytes in samples. RESULTS: The multianalyte immunosensor array had a throughput of 60 samples/h and allowed simultaneous detection of carcinoembryonic antigen, alpha-fetoprotein, beta-human choriogonadotropin, and carcinoma antigen 125 in clinical serum samples with concentrations up to 188 microg/L, 250 microg/L, 266 IU/L, and 334 kIU/L, respectively. The detection limits (limits of the blank, mean of blank plus 3 SD) were 1.1 micro/L, 1.7 microg/L, 1.2 IU/L, and 1.7 kIU/L. The inter- and intraassay imprecision (CVs) of the immunosensor arrays were <7.8% and <9.0%, respectively. The immunosensor arrays were stable for 28 days. CONCLUSIONS: This newly constructed immunosensor array provides a simple, automated, simultaneous multianalyte immunoassay with high throughput, short analytical time, and sufficiently low detection limits for clinical application. This method offers the capability of miniaturizing the multianalyte detection device.     

Cloning,expression, and purification of the hemolysin/cytolysin (HlyE antigen) from Salmonella enterica serovar Typhi: potential application for immunoassay development

The hemolysin (HlyE) protein of Salmonella enterica serovar Typhi was reported to be antigenic. This work describes the cloning, expression, and purification of a hexahistidine-tagged HlyE protein under native conditions. Immunoblot analysis and a competitive enzyme-linked immunosorbent assay using sera from typhoid patients showed the presence of HlyE-specific antibodies in circulation.     

全自动酶法测定血清总和游离卡尼汀浓度及临床应用

目的　建立自动化测定血清总和游离L 卡尼汀浓度的酶分光光度法 (CAT BTND)。方法　对卡尼汀酯水解条件及 pH和L 卡尼汀乙酰基转移酶浓度等反应条件作最佳化研究 ,同时对该法进行方法学评估 ,并用该法测定了 119名正常人的血清样本。结果　该法最低检出限为 4 81μmol/L ,线性范围可达 150 μmol/L(r =0 999)。批内和批间的平均CV分别为游离卡尼汀 5 52 %和5 90 % ,辛酯卡尼汀为 5 78%和 6 14 %。平均回收率为 98 5%和 98 4%。在胆红素≤ 12 0 μmol/L ,血红蛋白≤ 7g/L和抗坏血酸≤ 50 0 μmol/L的情况下 ,对测定结果无显著性影响。正常参考范围 :成人 :男 (3 2名 )总卡尼汀 (48 5± 17 7) μmol/L ,游离卡尼汀 (3 9 2± 14 2 ) μmol/L ,卡尼汀酯 (9 3±5 3 ) μmol/L ;女 (3 1名 )总卡尼汀 (42 7± 17 3 ) μmol/L ,游离卡尼汀 (3 2 4± 18 0 ) μmol/L ,卡尼汀酯(10 5± 5 5) μmol/L。儿童 (年龄 9～ 12岁 ) :男 (3 1名 )总卡尼汀 (57 3± 19 3 ) μmol/L ,游离卡尼汀(45 1± 14 4) μmol/L ,卡尼汀酯 (12 3± 10 4) μmol/L ;女 (2 5名 )总卡尼汀 (59 4± 15 7) μmol/L ,游离卡尼汀 (46 6± 12 6) μmol/L ,卡尼汀酯 (12 8± 7 8) μmol/L。 结论　该法正确快速 ,可作为L 卡尼汀缺乏症     

血清淀粉样蛋白A与C反应蛋白联合检测的临床应用价值

目的评估血清淀粉样蛋白A(SAA)与C反应的蛋白(CRP)联合检测的临床应用价值。方法采用胶体金法和免疫散射比浊法对82例感染性疾病组(包括15例肺炎、23例急性支气管炎、17例急性阑尾炎、16例急性胆囊炎、11例胰腺炎)患者及92名正常对照组的血清SAA和CRP水平进行检测,并比较两者间的浓度中位数及各疾病组的阳性检出率。结果感染性疾病组患者血清SAA与CRP浓度中位数水平分别为149.50 mg/L、34.20 mg/L,明显高于正常对照组分别为4.60 mg/L、3.82 mg/L,差异有统计学意义(P0.05);感染性疾病组血清SAA,与CRP阳性检出率分别为87.80%与79.26%。结论感染性疾病时,SAA与CRP水平变化趋势基本一致,且较CRP更为敏感。SAA与CRP都可作为一种监测炎症的有价值的标志物[1],SAA与CRP联合检测可以更有效的提高感染性疾病的检出率,为临床医生提供更敏感的诊断依据。     

甲状腺球蛋白抗体和微粒体抗体定量检测在自身免疫性甲状腺炎诊断中的应用

目的探讨甲状腺球蛋白抗体（TGAb）和甲状腺微粒体抗体（TMAb）定量榆测在自身免疫性甲状腺炎诊断中的应用。方法对219例甲状腺肿大患者和500例正常血清用酶免疫法（EIA）定苗检测甲状腺球蛋白抗体和甲状腺微粒体抗体。结果甲状腺肿大患者TGAb和TMAb定量检测结果分别为（295&#177;648）IU／mL和（309&#177;536）IU／mL,检出率分别为61％（134／219）和62％（136／219）,经过2种抗体的检测,最后有126例确诊为自身免疫性甲状腺炎（AITT）。500例健康对照TGAb和TMAb定量检测分别为（9&#177;4）IU/mL和（14&#177;6）IU／mL。结论EIA定量榆测血清TGAb和TMAb操作简便,结果准确,符合临床常规使用要求,对自身免疫性甲状腺炎的诊断、疗效观察具有重要临床应用价值。     

Relative immunogenicity of blood group antigens: First report in a Korean population

BackgroundThe immunogenicity of a blood group antigen is a measure of its likelihood of inducing alloantibodies. Although the immunogenicity of blood group antigens has been analyzed in Caucasian populations, immunogenicity to date has not been analyzed in Asian subjects. The present study therefore evaluated the relative immunogenicity of blood group antigens in a Korean population.Study design and methodsAll available data of unexpected antibody identification tests performed at Asan Medical Center between 1997 and 2021 were analyzed. The relative immunogenicity of a blood group antigen relative to K antigen was calculated based on relative numbers of alloantibodies and the probabilities of antigen-negative recipients receiving antigen-positive RBC units.ResultsA total of 3898 antibody identification results were included, with 1632 (41.9 %) from male patients. The ranking of antigen immunogenicity was: E > c > e > C > K > Jk(a) > Lu(a) > S > Fy(a) > Fy(b) > Jk(b) > Di(b) > Di(a) in the total population and E > c > e > C > Jk(a) > Fy(a) > Fy(b) > S > K > Lu(a) > Jk(b) > Di(b) > Di(a) in male patients.DiscussionThe rank order of immunogenicity for blood group antigens in this study provides information about relative immunogenecity in Koreans. These findings also provide supporting evidence regarding antigen selection for extended antigen-matched transfusions in recipients of multiple transfusions.     

酶抑制法测定m-AST试剂性能评价及其在肝病中的意义

目的评价酶抑制法测定天门冬氨酸氨基转移酶（AST）线粒体同工酶（m—AST）活性的性能,并探讨m—AST在肝脏疾病中的临床价值。方法采用酶抑制法检测m—AST活性[即采用蛋白酶完全抑制AST细胞质同工酶（c—AST）的活性,然后用速率法进行检测],并与免疫抑制法比较。采用酶抑制法检测259例肝病患者（包括急性肝炎43例、慢性病毒性肝炎95例、肝衰竭20例、重型肝炎11例、肝硬化代偿期40例、肝硬化失代偿期9例、原发性肝癌41例）及220名健康体检者（正常对照组）m—AST活性,并与AST比较。结果酶抑制法批内变异系数（CV）为0．59％～2．23％,批间CV为5．24％～6．23％;回收率为101．6％～108．0％,平均为104．97％;线性方程Y=0．997X一1．51,r=0．9999,m—AST活性在450U／L内线性良好;可完全抑制1500U／L的c—AST活性;与免疫抑制法结果呈高度正相关（r=0．9998）;溶血对m—AST检测结果干扰较大;以95％可信区间（孟±1．96s）初步确定m—AST参考区间男性为3．1～9．5U／L,女性为2．5～8．7U／L。肝病患者m—AST活性均高于正常对照组（P〈0．05）,并与AST呈正相关。正常对照组m—AST／AST比值为0．30±0．07,肝病患者m—AST／AST比值均低于正常对照组（P〈0．05）,但变化不如m—AST活性明显。结论酶抑制法测定m．AST活性自动化程度高,结果准确可靠,重复性好,线性范围宽。m—AST活性能反映肝细胞损伤的严重程度,对肝脏疾病的临床分类和预后具有重要价值。     

Antinuclear antibodies (ANA) screening by enzyme immunoassay with nuclear HEp-2 cell extract and recombinant antigens: analytical and clinical evaluation

OBJECTIVES: Immunofluorescence assay (IFA) has been the standard method for antinuclear antibodies (ANA). To simplify and standardize the ANA test, generic ANA solid phase enzyme immunoassay has been promoted. The objective of the present work has been to study the relationship with IFA and the clinical usefulness of a generic EIA for ANA (COBAS Core HEp-2 ANA EIA, Roche Diagnostics). DESIGNS AND METHODS: We studied 74 healthy individuals, 119 patients with defined systemic autoimmune diseases, 26 patients with other autoimmune diseases, and 490 routine samples sent to laboratory for ANA analysis. RESULTS: Precision study showed intra-assay coefficient of variations (CVs) below 8% and inter-assay CVs below 10%. In relation to IFA, a 0.6 kappa index of agreement was obtained. COBAS-ANA concentrations increased according to IFA titer and greatest COBAS-ANA responses were obtained with pure or mixed homogeneous patterns and centromeric patterns. Analysis of COBAS-ANA response to particular antigenic specificities showed that SS-B, Scl-70 and U1sn-RNP specificities were saturating at high concentrations, whereas Jo-1, SS-A and nuclear and centromeric specificities exhibited lower responses. Elevated serum concentrations of IgG and IgM did not interfere COBAS-ANA, but high serum rheumatoid factor (RF) concentrations produced a decrease of ANA. For systemic lupus erythematosus (SLE) patients, the COBAS-ANA best efficiency was obtained with a cut-off of 0.9, with a sensitivity of 97% and a specificity of 88%, whereas the best IFA-ANA efficiency was obtained with a 1:80 dilution, giving a sensitivity of 90% and a specificity of 99%. There were no differences between areas under ROC curves for COBAS-ANA and IFA-ANA. For other systemic and nonsystemic autoimmune diseases sensitivity and specificity of COBAS-ANA were similar or higher than that of 1:160 IFA-ANA titer. CONCLUSION: Sensitivity and specificity of COBAS Core ANA-EIA for SLE and other systemic and nonsystemic autoimmune diseases, together with performance characteristics make it an adequate automated system for ANA screening.     

MSCT与MRI在脑膜瘤诊断及手术入路评估中的临床应用价值

目的分析脑膜瘤的CT及MRI特征,评估CT及MRI在脑膜瘤术前定位、定性诊断及制定手术方案中的价值。方法回顾性分析经手术病理证实的27例脑膜瘤患者的CT及MRI资料,并与手术及术后病理结果对照。结果 27例脑膜瘤患者,MSCT平扫诊断率70.37%,MSCTA清晰显示肿瘤相邻的动脉与骨性解剖标志之间的三维关系;MRI平扫27例定位为颅内脑实质外肿瘤;MRI增强扫描诊断率96.30%,其中15例静脉窦旁脑膜瘤清晰显示肿瘤与静脉窦的相邻关系及静脉窦受压闭塞程度。依据MRI增强扫描及MSCTA定位诊断结果制定手术入路及方式,其中1例鞍旁脑膜瘤包绕颈内动脉虹吸部,另1例视神经管区域脑膜瘤包埋视神经部分予以残留;2例与矢状窦关系密切行脑膜瘤大部切除;余23例脑膜瘤完全切除。术后均无明确偏瘫、失明、失语等神经系统医源性损害症状。术后观察或随访1~24个月,无死亡病例。结论CT及MRI在脑膜瘤诊断、评估手术入路、保护肿瘤相邻动脉及静脉窦、减少医源性神经功能损害方面有重要价值。     

A modified method for the determination of Na-K-ATPase

A modified method for determination of adenosine triphosphatase (ATPase) activities, with special reference to Na-K-ATPase, in human erythrocytes using 2 mmol/l ethylenediamine tetraacetic acid in the hemolyzing solution is presented. This modification gives stable day to day activities with a good power of discrimination. Small blood volumes and only standard laboratory equipment are needed.     

血清酶测定结果可溯源性研究

目的　通过对酶标准品的测定 ,探讨血清酶测定结果的可溯源性。方法　向参加研究的实验室 (50家 )各发放酶校准品 1支 ,各实验室用常规方法测定校准品 ,记录检验结果 ;计算校准品测定结果总体均值与标示值间的偏倚。结果　丙氨酸氨基转移酶 (ALT)、天门冬氨酸氨基转移酶(AST)、肌酸激酶 (CK)和乳酸脱氢酶 (LD)的测定结果均值与标准品标示值的相对偏差≤± 10 % ,分别为 6 3 % ,5 5% ,-5 9%和 -5 0 % ;碱性磷酸酶 (ALP)的实验室检测结果与标准品标示值间的相对偏倚为 -3 6 6% ,淀粉酶测定结果的实验室间CV为 2 9 2 %。结论　我国临床实验室对血清酶的测定结果有向国际标准溯源的基础