免疫印迹法检测老年2型糖尿病患者血清胰岛自身抗体的临床价值

目的 评价免疫印迹法检测老年2型糖尿病患者胰岛自身抗体(IAA)的临床价值. 方法 采用免疫印迹(IB)法和酶联免疫吸附(ELISA)法分别检测350例老年2型糖尿病患者和120例健康对照组血清谷氨酸脱羧酶抗体(GADA)、胰岛细胞抗体(ICA)和IAA,比较不同方法检测胰岛相关抗体的阳性率. 结果 在老年2型糖尿病患者中,IB法检测GADA阳性患者为56例,ELISA法检测阳性患者为38例,IB法检测的阳性率要明显高于ELISA法,而2种方法检测ICA、IAA的阳性率比较无明显差异.2种方法检测病程≥5年患者GADA、ICA的阳性率均低于病程＜5年患者,但IB法检测GADA的阳性率要高于ELISA法. 结论 IB法在检测老年2型糖尿病患者IAA尤其是GADA的敏感性要优于ELISA法.     

免疫人群乙肝抗体阳性率调查及乙肝核心抗体检测方法探讨

目的 调查我院检测人群乙型肝炎病毒表面抗体(HBsAb)及乙型肝炎病毒核心抗体(HBcAb)阳性率,并探讨检测方法选择及检测操作模式对免疫人群乙肝核心抗体结果的影响。方法 统计2012—2013年用化学发光微粒子免疫检测法(CMIA)和酶联免疫竞争抑制一步法(ELISA)检测乙肝人群的HBsAb及HBcAb阳性率,并按≤2岁、2~20岁、>20岁3个年龄分段比较;收集92例免疫人群样本用CMIA和3种ELISA法检测HBcAb;用同种ELISA法试剂,改变操作模式进行HBcAb检测。结果 3组年龄段检测人群HBsAb两种检测方法统计的阳性率无统计学差异,≤2岁组阳性率最高;HBcAb用CMIA法统计的阳性率在≤2岁组和2~20岁组的免疫人群组低于ELISA法。 92例样本用CMIA法检测HBcAb阳性率为2.2%;用3种ELISA法试剂检测阳性率分别为79.3%、82.6%及94.6%;3种ELISA法试剂检测阳性率无统计学差异。92例样本改变操作模式,用同种ELISA法检测试剂检测结果有19例为阴性,差异有统计学意义。结论 检测人群HBsAb检出率在2岁及以下人群最高,随年龄增长抗体下降。 HBcAb用CMIA法检测在免疫人群中检出阳性率明显低于EIISA法。ELISA竞争抑制一步法检测HBcAb易造成假阳性,如改用HBcAb单项加样操作或选用化学发光微粒子免疫检测法(CMIA)检测,可明显减少假阳性。     

不同血型成人胰腺用于ICA检测的比较研究

目的应用免疫组化法(ABC法)检测ICA,比较研究A型、B型、AB型与O型4种不同血型成人胰腺抗原底物对ICA检测的特异性和敏感性的影响。方法取24例1型糖尿病病人的ICA阳性不同JDF单位的血清标本和23例正常人ICA阴性血清标本,以A型、B型、AB型与O型4种不同血型成人胰腺的冰冻切片为抗原底物,应用ABC免疫组化法检测ICA。结果以O型血成人胰腺为对照,应用3例A型和B型血胰腺和2例AB型血胰腺为抗原底物进行ICA检测,24例ICA阳性血清和23例ICA阴性血清检测结果的特异性和敏感性基本一致,未见假阳性或假阴性结果。结论本文研究结果证明A型、B型或AB型血成人胰腺与O型血成人胰腺一样,可用于ICA免疫组化(ABC法)检测,不同血型人胰腺作为抗原底物不影响ICA免疫组化法检测的特异性和敏感性。     

金标法初筛试剂与诊断试剂检测乙肝表面抗原的结果分析

目的:分析金标法初筛试剂与诊断试剂检测无偿献血者乙肝表面抗原的阳性率、假阳性率及假阴性率的结果。方法:金标法初筛试纸条检测68 326例,诊断试纸条检测55 637例,乙肝表面抗原初检阴性者与初检阳性者均用ELISA法做复检。结果:初筛试剂检测68 326例的HBsAg阳性率为5.96%、假阳性率5.8%、假阴性率2.84%,诊断试剂检测55 637例的阳性率为2.77%、假阳性率2.6%、假阴性率0.47%。结论:金标法初筛试剂检测无偿献血者乙肝表面抗原的阳性率、假阳性率及假阴性率均高于诊断试剂。     

免疫金银染色法、斑点ELISA和斑点免疫金银染色法检测华支睾吸虫病人血清中抗体的研究

用免疫金银染色法(IGSS)、斑点ELISA和斑点免疫金银染色法(Dot-IGSS)三种方法,同时检测40份华支睾吸虫病人血清,阳性率分别为100％、90％和95％,检测40份正常人血清,阴性率分别为100％、97.5％和97.5％,显示三种方法均有较高的敏感性和特异性。     

基因芯片技术检测慢性乙型肝炎肝硬化患者肝组织及血清HBVDNA研究

目的研究DNA芯片技术在检测乙型肝炎和肝硬化患者肝组织及血清中HBVDNA的应用，并与雅培试剂、免疫组织化学和原位分子杂交法比较。方法用点样仪将PCR扩增的HBVDNA探针制成基因芯片。对15例慢性乙型肝炎(下称慢乙肝)患者的血清及肝活检组织、99例乙型肝炎后肝硬化肝组织，分别用基因芯片、原位分子杂交法、免疫组织化学法、雅培试剂检测HBVDNA、HBcAg、HBsAg、HBeAg。结果15例慢乙肝HBsAg、HBeAg阳性患者的乙型肝炎血清基因芯片检测均阳性。15例肝组织标本中，免疫组化法HBcAg阳性15例，HBVDNA原位分子杂交阳性14例。基因芯片检测阳性14例。99例乙型肝炎后肝硬化肝组织标本中，HBcAg阳性67例、HBVDNA阳性53例，基因芯片检测阳性46例；32例HBcAg、HBVDNA阴性组织基因芯片检测均阴性。结论肝炎基因诊断芯片可以检测肝组织及血清HBVDNA，其诊断准确率高，假阳性率低。     

ELISA和TRUST检测梅毒螺旋体方法比较分析

目的 比较ELISA(酶联免疫吸附试验)、TRUST(甲苯胺红不加热血清试验)法诊断梅毒螺旋体感染的方法学差异.方法 用目前国内最常用的诊断梅毒螺旋体感染的TRUST试剂及ELISA试剂检测722例标本,同时与TPPA(梅毒螺旋体明胶凝集试验)的检测结果进行比较,从而得到各试验的假阴性率和假阳性率.结果 对722份样本的检测中,ELISA和TPPA的阳性符合率为98.26％,假阴性率和假阳性率分别为0.46％和1.91％,2种方法差异无统计学意义(P ＞0.05); TRUST和TPPA的阳性符合率为84.39％,假阴性率和假阳性率分别为21.08％和5.10％;ELISA和TURST检出率差异有统计学意义(P＜0.01).结论 ELISA测定梅毒螺旋体的方法在日常大量标本检验时优于TRUST.     

胶体金免疫层析试纸条对云南鼠疫现场材料的检测

目的评价检测鼠疫抗原及抗体胶体金免疫层析试纸条(G ICA及RG ICA)对现场材料的检测效果。方法采用G ICA分别对采自云南省的607份血清标本(查鼠疫抗体)及572份鼠脏器标本(查鼠疫抗原)进行检测,同时以血凝试验(IHA及R IHA)与酶联试验(ELISA及F1-ELISA)作为对照。结果(1)在对607份血清标本的检测中,G ICA、IHA及ELISA三种方法的符合率中度,而G ICA的敏感性比IHA与ELISA分别高111%和90%;(2)在对572份鼠脏器标本的检测中,RG ICA、R IHA及F1-ELISA三种方法的符合率中度,而RG ICA的敏感性比R IHA与F1-ELISA分别高100%和14.3%。结论检测鼠疫的G ICA及RG ICA特异性强、灵敏度高、简便快速,有较大的推广应用价值。     

检测鼠疫抗体胶体金免疫层析试纸条的研制

目的建立快速、简易的检测鼠疫F1抗体的胶体金免疫层析法(G ICA)。方法(1)采用胶体金颗粒标记纯化F1抗原,并将标记物喷于玻璃纤维;同时将纯化F1抗原喷线固定于硝酸纤维素膜上,用于F1抗体的捕捉;按常规组装成检测鼠疫抗体免疫层析试纸条。(2)采用该试纸条与血凝法对同一份兔抗F1抗体进行检测,以评价该试纸条的敏感性。(3)采用该试纸条对44株非鼠疫菌的免疫鼠血清进行检测,以评价该试纸条的特异性。(4)采用该试纸条、血凝法及ELISA对607份血清标本进行检测,以评价该试纸条对现场材料的检测效果。结果(1)该试纸条可在15 m in之内完成检测;(2)在敏感性上,该试纸条对同一份免疫兔血清的检测较血凝法高一个滴度;(3)对被试的44株所选菌株的免疫鼠血清的检测均为阴性;(4)在对607份血清标本的检测中,免疫层析试纸条、血凝及ELISA三种方法的符合率中度,而免疫层析试纸条的敏感性分别比血凝与ELISA高111%和90%。结论以纯化的鼠疫F1抗原为基础建立的G ICA检测鼠疫F1抗体的方法特异性强、灵敏度高、简便快速,无需特殊仪器设备,有较大的推广应用价值。     

免疫印迹法检测胰岛自身抗体系列在糖尿病分型诊断中的应用

目的探讨免疫印迹法检测胰岛自身抗体系列——抗谷氨酸脱羧酶抗体(GADA)、抗胰岛细胞抗体(ICA)、抗胰岛素自身抗体(IAA)在糖尿病分型诊断中的应用及临床意义。方法应用免疫印迹法联合检测187例糖尿病患者及30例健康体检者血清中的GADA、ICA、IAA。结果应用免疫印迹法检测T1DM组中GADA、ICA、IAA阳性率分别为65.22%、53.62%和28.99%,显著高于T2DM组(P<0.05)及正常对照组(P<0.05);T2DM组GADA、ICA、IAA阳性率分别为12.71%、11.02%和5.93%,与正常对照组比较有统计学差异(P<0.05)。结论免疫印迹法检测胰岛自身抗体系列对于临床糖尿病的诊断及分型具有重要的临床应用价值。     

检测华支睾吸虫特异性IgG4生物素-亲和素复合酶联免疫吸附法的建立及检测效能分析

目的探讨亲和素-生物素复合酶联免疫吸附法(ABC-ELISA)检测特异性Ig G4抗体用于诊断华支睾吸虫病的效果。方法建立检测华支睾吸虫特异性抗体Ig G4的ABC-ELISA法(Ig G4-ABC-ELISA),以此方法检测华支睾吸虫病、日本血吸虫病、卫氏并殖吸虫病、弓形虫病、棘球蚴病、囊尾蚴病和曼氏裂头蚴病患者血清样本。以Ig G4-ELISA和Ig GELISA法为对照,比较3种方法用于诊断华支睾吸虫病的敏感性、特异性、阳性预测值、阴性预测值及诊断效率。结果成功建立了用于检测华支睾吸虫特异性抗体的Ig G4-ABC-ELISA法,用此方法检测华支睾吸虫病患者血清特异性抗体Ig G4的敏感性为90.0%,特异性为98.2%,阳性预测值为93.8%,阴性预测值为97.0%,诊断效率为96.3%;Ig G4-ELISA法检测华支睾吸虫病患者血清特异性抗体敏感性为86.0%,特异性为98.2%,阳性预测值为93.5%,阴性预测值为95.9%,诊断效率为95.4%;Ig G-ELISA法检测华支睾吸虫病患者血清特异性抗体Ig G的敏感性为94.0%,特异性为88.1%,阳性预测值为70.1%,阴性预测值为98.0%,诊断效率为89.4%。Ig G4-ABC-ELISA法检测华支睾吸虫病患者血清特异性抗体的敏感性高于Ig G4-ELISA法(P0.05),特异性高于Ig G-ELISA法(P0.05)。结论 Ig G4-ABC-ELISA法检测华支睾吸虫特异性抗体IgG4具有高度敏感性与特异性,在华支睾吸虫病诊断中具有较好应用价值。     

A Comparison of Line Blots,Enzyme-linked Immunosorbent,and RNA-immunoprecipitation Assays of Antisynthetase Antibodies in Serum Samples from 44 Patients

Objective To determine the differences between anti-aminoacyl tRNA synthetase (ARS) antibodies among line blots, enzyme-linked immunosorbent assay (ELISA) anti-ARS tests, and RNA-immunoprecipitation (IP) assays. Methods Sera from patients with confirmed or suspected antisynthetase syndrome (ASS) that were positive for either the anti-ARS test or the line-blot assay were used to perform an RNA-IP assay and ELISA to detect individual anti-ARS antibodies. Results Among the 44 patients, 10 were positive only in line-blot assays, 6 were positive only in the anti-ARS test, and 28 were positive in both assays. We compared the accuracy of these assays against the gold standard RNA-IP assay. The κ coefficient was 0.23 in the line-blot assay, but this increased to 0.75 when the cut-off was increased from 1+ to 2+. The κ coefficient was 0.73 in the anti-ARS test. The κ coefficient was 0.85 for positivity in both assays. Patients with ASS that was positive in an RNA-IP assay more frequently had mechanic''s hand (62.1% vs. 20%: p=0.031), myositis (51.7 vs. 10%: p=0.028) and more ASS symptoms than those who were positive only in line-blot assays (3.48 vs. 2.2: p=0.019). Conclusions Clinicians need to understand the features of each assay and determine diagnoses by also considering clinical presentations. Diagnoses should not be judged based only on the results of line-blot assays due to the risk of a misdiagnosis from false positives.     

ABC-ELISA法诊断肠螨病的研究

目的　探讨生物素 -亲和素酶联免疫吸附试验 (ABC -ELISA)在肠螨病诊断中的应用价值。方法　采集 4 8例肠螨病患者血清 ,用ABC -ELISA法检测血清中螨特异性抗体IgG ,并与葡萄球菌A -蛋白酶联免疫吸附试验 (SPA -ELISA)进行比较。结果　用ABC -ELISA法和SPA -ELISA法检测 4 8例肠螨病患者血清中的螨特异性抗体IgG阳性率分别为89 5 8% (43/ 4 8)和 5 6 2 5 % (2 7/ 4 8) ,两者比较 ,差异具显著性 (χ2 =13 5 0 ,P <0 0 1)。结论　ABC -ELISA法是实验室诊断肠螨病的一种有效方法     

谷胱甘肽-S-转移酶多克隆抗体免疫金测定法检测日本血吸虫循环抗原的研究

目的探讨谷胱甘肽－Ｓ－转移酶（ｒＳｊ２６ＧＳＴ）多克隆抗体诊断日本血吸虫病的应用价值。方法以抗ｒＳｊ２６ＧＳＴ多克隆抗体力捕获并检测抗体，建立双抗体夹心斑点免疫金测定技术，用于检测日本血吸虫循环抗原，并以斑点免疫金测定法和ＥＬＩＳＡ检测抗体作对照。结果３种方法检测急性血吸虫病的敏感性均为１００％；对慢性血吸虫病的敏感性分别为７６．４％、９８．１％和９６．２％；特异性分别为９５．０％、９８．０％和９６．０％。结论ｒＳｊ２６ＧＳＴ抗原对日本血吸虫病具有诊断价值，可望有较好的应用前景。     

A new ELISA assay for diagnosis of acquired von Willebrand Syndrome

The pathophysiology of acquired von Willebrand syndrome (AVWS), a rare bleeding disorder, is not fully understood. Circulating antibodies to Von Willebrand factor (VWF) are found in patients with AVWS associated with lymphoproliferative disorders but these autoantibodies are difficult to detect with routine laboratory tests and neutralisation assays. We have developed a simple enzyme-linked immunosorbent assay (ELISA) to detect serum antibody binding to VWF protein immobilized on polystyrene plates. Ten patients with AVWS were studied, eight of whom also had lymphoproliferative disorders. We found antibodies in eight patients; all of them were positive for IgG and five were also positive for IgM. This simple method appears to be more sensitive than functional assays, which failed to identify two of the patients who were positive with the ELISA. In conjunction with other tests, this ELISA method may be useful for demonstrating the immunological mechanism underlying some cases of AVWS. Such patients would qualify for intravenous immunoglobulin therapy, which can correct the clotting disorder.     

Autoantibodies to M2 mitochondrial autoantigens in normal human sera by immunofluorescence and novel assays

Primary biliary cirrhosis (PBC) is characterized by the presence of antimitochondrial antibodies (anti-M2), directed against the E2 subunits of the 2-oxo-acid dehydrogenase complexes (2-OADC), chiefly pyruvate dehydrogenase complex (PDC-E2). We present here a detailed study, based on a large panel of normal sera, of the specificity of tests for anti-M2 by immunofluorescence and for anti-PDC by other assays for the diagnosis of PBC. The assays for anti-PDC included immunoblotting with bovine heart mitochondria, ELISA using recombinant PDC-E2 and an enzyme inhibition assay using purified porcine PDC. The positivity rates for normal sera were 0 (0/170), 2 (4/201), 1.5 (3/198) and 0% (0/186) for immunofluorescence, immunoblotting, ELISA and the enzyme inhibition assay, respectively. The seven positive reactions detected either by immunoblotting (n= 4) or ELISA (n= 3) were negative by the other three assays and in no instance did biochemical indices give any indication of chronic liver disease. Thus, as judged by reactivity with normal sera, the specificity of a positive test for the antibody to the major M2 autoantigen (PDC-E2) is 100% for immunofluorescence and the enzyme inhibition assay, 98% for immunoblotting and 98.5% for ELISA.     

胰岛细胞抗体ABC法检测及临床意义初步探讨

胰岛素依赖型糖尿病（ＩＤＤＭ）与机体的免疫功能紊乱有密切的关系。胰岛细胞抗体（ＩＣＡ）是ＩＤＤＭ患者主要的免疫学标志之一。可采用免疫组织化学技术作ＩＣＡ定性检测。我们首次应用“Ｏ”型血正常人胰腺石蜡切片作为抗原，ＡＢＣ法（卵白素－生物素化过氧化物酶复合物法）检测血清中ＩＣＡ。结果：１７例ＩＤＤＭ患者，阳性检出率５２．９４％；２０例ＮＩＤＤＭ及２０例非糖尿病患者无阳性反应。与国外报道比较，其方法的可     

Comparison of two second-generation anti-hepatitis C virus ELISA on 21431 US blood donor samples

SUMMARY. We have compared two different second-generation (2.O) enzyme-linked immunosorbent assays (ELISA) for the presence of antibodies to hepatitis C virus (anti-HCV) in blood from volunteer, unpaid donors. At two separate blood centres, a total of 21431 donor samples were tested with Abbott Anti-HCV 2.0 ELISA and Ortho Anti-HCV 2.0 ELISA. Samples found to be repeatedly reactive were tested by supple-mental/investigational assays, MATRIX HCV (Abbott) and anti-HCV RIBA II (Ortho/Chiron), to 'confirm' the presence of anti-HCV. Discordant ELISA samples were additionally tested by the polymerase chain reaction (PCR) for the presence of HCV RNA. The Abbott anti-HCV assay had a repeatedly reactive rate of 0.59% (127/21431) and the Ortho anti-HCV assay 0.51% (110/21431). Overall agreement between assays was 99.76%. 72/127 (56.7%) of Abbott repeatedly reactive samples confirmed on MATRIX and 61/127 (48.0%) on RIBAII; 70/110 (63.6%) of Ortho repeatedly reactivate samples confirmed on MATRIX and 61/110 (55.5%) on RIBA II. Discordant ELISA samples tested by PCR yielded negative results. Hence the two ELISA had equal sensitivity, as defined by detection of true positive samples: the slightly lower specificity of the Abbott Anti-HCV 2.0 ELISA may be owing to culling of donors with a false positive test by Ortho's Anti-HCV 1.0 and 2.0 ELISA tests (the routine tests in place at each blood centre). A sample found to be repeatedly reactive by two different ELISA tests for anti-HCV is likely to be a true positive and may not require further 'confirmatory' testing.