快速胶体硒法检测抗-HIV(1+2)抗体

人类免疫缺陷病毒 (HIV)抗体测定是鉴别HIV病毒感染血液的最合适指标 ,检测HIV抗体最常用的方法有三种 :酶联免疫吸附试验 (ELISA)、颗粒凝集试验、特异性快速试验。特异性快速试验方法简单、快速、不需特殊仪器。本文对HIV抗体胶体硒法试剂检测抗 -HIV( 1 +2 )抗体的灵敏度、特异性、反应时间做了初步探讨 ,现报告如下 :1　材料与方法1 .1　检测参考品HIV抗体国家参考品 (中国药品生物制品检定所 ,批号 :2 0 0 1 0 7)。该参考品有 45支样品组成 ,其中N1～N2 0 为HIV抗体阴性参考品 ,P1～P18为HIV-1型阳性样品 ,P19～P2 0 为…     

新疆伊宁县婚前检查者人类免疫缺陷病毒HIV(1+2型)抗体感染情况分析

来医院婚前1332名检查者自然人群静脉采血2～3ml分离血清后进行HIV抗体检测时发现其中的15例HIV抗体阳性情况,现报告其血清学检测结果并对这一结果出现的原因做出初步分析。     

HIV(1+2)抗体/P24抗原联合法与胶体硒法在临床检测HIV中的应用研究

目的比较第4代酶联免疫吸附测定(ELISA)[HIV(1+2)抗体与P24抗原联合检测]和胶体硒法在临床检测HIV中的特点。方法分别用第4代ELISA检测试剂和胶体硒试剂检测HIV抗体,初筛阳性样本送重庆市疾病预防控制中心进行Western blot确证试验。结果第4代ELISA试剂检测、胶体硒检测及Western blot检测HIV的阳性率分别为0.34%、0.29%及0.27%。第4代ELISA的HIV漏检率(1.3%)低于胶体硒法(4.9%),而其检测HIV的假阳性率(0.03%)高于胶体硒试剂(0.01%)。结论第4代ELISA和胶体硒法联合应用可提高HIV检测的准确性。     

胶体金免疫层析法在早孕检测中的应用

目的:免疫层析法是将特异的抗体先固定于硝酸纤维素膜的某一区带,当该干燥的硝酸纤维素膜一端浸入样品(尿液或血清)后,由于毛细管作用,样品沿该膜向前移动,当移动至固定有抗体的区域时,样品中相应的抗原与该抗体发生特异性结合,若用免疫胶体金染色可使该区域显色。胶体金标记稳定、易贮存,常用于定性或半定量的快速免疫检测方法中.在免疫层析快速诊断技术中常用于β-HCG的特异性免疫诊断早孕检测等,具有快速、准确和无污染等优点。     

胶体金检测抗环瓜氨酸肽抗体性能评价

目的对胶体金免疫层析（GICA）技术检测患者血清抗环瓜氨酸肽（CCP）抗体进行性能验证和评价。方法采用酶联免疫吸附试验（ELISA）和GICA,同时对104例可疑类风湿性关节炎（RA）患者进行抗-CCP抗体检测,计算两种方法检测结果的阴性符合率、阳性符合率、总符合率以及Kappa值。用±20％临界值浓度标本各重复检测20次,记录阴性和阳性结果数,验证GICA法重复性。结果两种方法检测结果比较,阴性符合率、阳性符合率、总符合率、Kappa值分别为92％、100％、96％、0．92。GICA法检测＋20％临界值浓度标本结果阳性数为20次,阳性率100％;-20％临界值浓度标本结果阴性数为15次,阴性率为75％。结论GICA法与ELISA法检测抗-CCP抗体结果一致性良好,而且GICA方法简单、快速,敏感性和特异性较高,是早期筛查类风湿性关节炎值得推广的方法。     

胶体金免疫层析法检测梅毒螺旋体特异性抗体的方法学评价

目地 :对胶体金免疫层析法检测梅毒螺旋体特异性抗体进行方法学评价。方法 :采用 GICA法 RPR法 TPPA法对2 2 9例血清样品同时测定 ,并结合物理现象作观察 ,对试条的敏感度、特异度、渗透速度、固相膜均一性等进行评价。结果 :GICA检测不同期梅毒的优缺点同当今使用的方法一致     

胶体金免疫层析法检测血清β-HCG的性能评价

目的 评价胶体金免疫层析法"早早孕"试纸条检测血清中人绒毛膜促性腺激素β亚单位(β-HCG)的性能.方法 通过与化学发光法比较,从灵敏度、特异度、重复性、检出限等方面评价杭州艾康生物技术公司生产的"早早孕"人绒毛膜促性腺激素试纸条(胶体金免疫层析法).结果 "早早孕"试纸条检测血清β-HCG的最低检出限为25 mIU/mL,以化学发光法为真值,其灵敏度为84.6%,特异性为100%,尤登指数为84.6%,准确度为93.3%.试纸条浸入标本的最佳时间为10 s,观察结果最佳时间为3 min,检测结果在10 min之内稳定.该方法重复性良好,实验不受溶血、脂血的干扰.结论 杭州艾康生物技术公司生产的"早早孕"试纸条(胶体金免疫层析法)检测血清β-HCG的特异度高,重复性好,准确度高.     

ELISA法与金标层析法检测HIV抗体的适用性探讨

目的将金标层析法与ELISA法检测HIV—Ab的结果进行分析比对，以探讨两种方法检测HIV抗体在日常工作中的适用性。方法对1208例血液样本用金标层析法与ELISA法检测其HIV抗体。结果两种方法检测HIV抗体的结果差异无显著性，ELISA法的特异性高于金标层析法。结论两种方法均可适用于医学检测，金标层析法适合于急诊、流动采血点、基层单位的推广使用，而ELISA法适合于大中型医院HIV抗体筛查和艾滋病流行病学监测等。     

ELISA法初筛献血员anti-HIV(1/2型)初探

我科用4种不同厂家生产的酶联免疫吸附(ELISA)试剂2 277人次献血员初筛anti-HIV(1/2型)抗体,结果报道如下. 1 材料与方法 1.1 标本的采集与处理 于献血前采集小标本,空腹肘静脉血2ml,不抗凝;采血后留取2ml不抗凝标本.标本充分吸收后,离心得到血清,以消除纤维蛋白的干扰.检测在4h内进行完毕.     

创伤弧菌快速检测试纸条的研制

摘要：目的研制一种敏感、特异、快速的免疫试纸条,用于创伤弧菌检测。方法用创伤弧菌（ATCC27562）菌体抗原免疫
家兔,制备兔抗创伤弧菌多克隆抗体;采用柠檬酸盐还原法制备胶体金颗粒并标记多克隆抗体,纯化后滴加在玻璃纤维素
膜上;在硝酸纤维素膜上包被兔抗创伤弧菌多克隆抗体和羊抗兔IgG,分别作为检测线和质控线;将处理后的硝酸纤维膜、
玻璃纤维素膜与样品垫、吸水纸组装成试纸条,4 ℃保存并定期检测;设阳性及阴性对照,对本试纸条检测的灵敏度、特异
性进行检测。结果将试纸条插入样品溶液中,20~30 min内即可显示结果,其检测灵敏度为2×106 CFU/ml;特异性检测显
示本试纸条与溶血性葡萄球菌、金黄色葡萄球菌、副溶血弧菌等12种菌株均无交叉反应;稳定性检测表明该试纸条可4 ℃保
存4个月。结论本研究构建出创伤弧菌多克隆检测试纸条,具有较高的灵敏度和较好的特异性,简便,20~30 min即可获
得检测结果,可用于创伤弧菌的快速检测。     

Development of novel-nanobody-based lateral-flow immunochromatographic strip test for rapid detection of recombinant human interferon α2b

Recombinant human interferon α2b (rhIFNα2b) is widely used as an antiviral therapy agent for the treatment of hepatitis B and hepatitis C. The current identification test for rhIFNα2b is complex. In this study, an anti-rhIFNα2b nanobody was discovered and used for the development of a rapid lateral flow strip for the identification of rhIFNα2b. RhIFNα2b was used to immunize an alpaca, which established a phage nanobody library. After five steps of enrichment, the nanobody I22, which specifically bound rhIFNα2b, was isolated and inserted into the prokaryotic expression vector pET28a. After subsequent purification, the physicochemical properties of the nanobody were determined. A semiquantitative detection and rapid identification assay of rhIFNα2b was developed using this novel nanobody. To develop a rapid test, the nanobody I22 was coupled with a colloidal gold to produce lateral-flow test strips. The developed rhIFNα2b detection assay had a limit of detection of 1 μg/mL. The isolation of I22 and successful construction of a lateral-flow immunochromatographic test strip demonstrated the feasibility of performing ligand-binding assays on a lateral-flow test strip using recombinant protein products. The principle of this novel assay is generally applicable for the rapid testing of other commercial products, with a great potential for routine use in detecting counterfeit recombinant protein products.     

抗β2GPI抗体测定在系统性红斑狼疮患者中的临床意义

目的探讨抗β2糖蛋白I抗体(aβ2GPI)在系统性红斑狼疮(SLE)诊治中的临床意义及与疾病活动性的关系。方法采用酶联免疫吸附法(ELISA)测定68例SLE患者及23例健康对照者血清中aβ2GPI浓度,同时记录SLE患者各项实验室指标和其他自身抗体的水平。对照分析aβ2GPI阳性与阴性患者实验室检测指标结果的差异。结果SLE患者中,aβ2GPI抗体阳性率为25%(17/68),健康对照组中未检出aβ2GPI。患者的SLE疾病活动性指数(SLEDAI)与aβ2GPI抗体浓度无相关性。aβ2GPI阳性SLE患者血小板减少、凝血酶原时间(PT)延长的发生率高于aβ2GPI阴性组(<0.05)。aβ2GPI检测结果与aCL检测结果间存在正相关性。结论部分SLE病例中可检出抗β2GPI抗体。抗β2GPI抗体浓度与SLE疾病活动性无明显相关。抗β2GPI抗体与SLE患者发生血小板减少、凝血酶原时间延长等事件相关。     

肿瘤及对照人群血清细小病毒H1抗体检测

细小病毒能抑制自发和诱发的肿瘤形成，具有明显的抗肿瘤作用。人群血清流行病学研究表明细小病毒ＡＡＶ感染和宫颈癌的低发生率有关。本文用血凝抑制试验检测肿瘤患者乾和健康对照人群血清细小病毒Ｈ１抗体。结果表明，４１４例肿瘤患者血清中３例抗细小病毒Ｈ１抗体阳性（０．７５％），对照血清４２０例中有２１例阳性（５．０％）。肿瘤患者血清细小病毒Ｈ１抗体阳性率明显低于对照人群（Ｐ〈０．００１）。提示细小病毒Ｈ１感染     

Rapid immunochromatographic strip test for the detection of albuminuria and brief literature review on albuminuria screening

Albuminuria is a powerful predictor of cardiovascular events in diabetic and hypertensive patients as well as in the general population. In the present study we evaluated the diagnostic performance of the new PreventID Albumin test, a semi-quantitative immunochromatographic dipstick device for the rapid detection of low concentrations of urinary albumin (microalbuminuria). 88 randomly selected fresh urine samples from the central clinical laboratory of the Charité, Berlin/Germany were analysed. The diagnostic accuracy of the PreventID Albumin test for the detection of urinary albumin excretion >18 mg/l was assessed by comparing the results with urinary albumin excretion determined by immunoturbidimetry as golden standard. In comparison with immunoturbidimetry the PreventID Albumin test had a sensitivity of 96.2% and specificity of 97.1%. False negative results were found in 2.3% and false positive results were obtained in 1.1% of specimens. These findings suggest that the PreventID Albumin test may be a useful and valid method for the screening of albuminuria. However, it should not be regarded as a diagnostic test. Positive results should be followed by quantification of urinary albumin or albumin/creatinine ratio by a laboratory based method.     

Usefulness of enzyme immunoassay (EIA) for screening of anti HIV antibodies in urinary specimens: A comparative analysis

Background

Standard HIV testing is done using serum or plasma. FDA approved ELISA to screen urine for IgG antibodies to HIV-1 in 1996. It is a simple, noninvasive test and is appropriate for developing countries where health care personnel may not be professionally trained or where clean needles for drawing blood may not always be available.

Methods

436 individuals with high-risk behavior and strong clinical suspicion of HIV infection were screened for IgG antibodies to HIV-1 in urine by ELISA. Urine HIV testing was performed by enzyme immunoassay, at the ongoing Voluntary Confidential Counseling and Testing Center (VCCTC) at a large tertiary care microbiology lab. The individuals enrolled for the study had high-risk exposure to the virus and majorities were from a state with a high incidence of HIV infection. In all individuals, both serum and urine were tested for IgG antibodies to HIV-1.

Results

Overall, 135 individuals (30.96%) were HIV-positive, of whom 96 (71%) had never previously tested positive; 87% of those who tested positive received their results, and most were referred for medical care. Sensitivity, specificity and predictive values of HIV-1 urine ELISA test kit were determined. Sensitivity was found to be 89.6%; 95% CI [82.9–94.0], specificity 97.3%; 95% CI [94.6–98.8], positive predictive value 93.8%; 95% CI [87.8–97.1] and negative predictive value 95.4%; 95% CI [92.3–97.4].

Conclusion

Efficiency, sensitivity, and specificity of the urine-based screening for HIV-1 test kits were excellent as compared to the reference test.     

Usefulness of enzyme immunoassay (EIA) for screening of anti HIV antibodies in urinary specimens: A comparative analysis

Background

Standard HIV testing is done using serum or plasma. FDA approved ELISA to screen urine for IgG antibodies to HIV-1 in 1996. It is a simple, noninvasive test and is appropriate for developing countries where health care personnel may not be professionally trained or where clean needles for drawing blood may not always be available.

Methods

436 individuals with high-risk behavior and strong clinical suspicion of HIV infection were screened for IgG antibodies to HIV-1 in urine by ELISA. Urine HIV testing was performed by enzyme immunoassay, at the ongoing Voluntary Confidential Counseling and Testing Center (VCCTC) at a large tertiary care microbiology lab. The individuals enrolled for the study had high-risk exposure to the virus and majorities were from a state with a high incidence of HIV infection. In all individuals, both serum and urine were tested for IgG antibodies to HIV-1.

Results

Overall, 135 individuals (30.96%) were HIV-positive, of whom 96 (71%) had never previously tested positive; 87% of those who tested positive received their results, and most were referred for medical care. Sensitivity, specificity and predictive values of HIV-1 urine ELISA test kit were determined. Sensitivity was found to be 89.6%; 95% CI [82.9–94.0], specificity 97.3%; 95% CI [94.6–98.8], positive predictive value 93.8%; 95% CI [87.8–97.1] and negative predictive value 95.4%; 95% CI [92.3–97.4].

Conclusion

Efficiency, sensitivity, and specificity of the urine-based screening for HIV-1 test kits were excellent as compared to the reference test.     

抗中性粒细胞胞浆抗体3种检测方法的比较

目的:寻找敏感性、特异性较高又适宜我国目前推广、普及的抗中性粒细胞胞浆抗体(antineutrophilcytoplasmicantibodies,ANCA)的检测方法。方法:对598份血清应用间接免疫荧光(indirectimmunofluorescence,IIF)、总抗原ELISA、抗原特异性ELISA(MPO和PR3ELISA)等3种方法进行ANCA检测。结果:IIF法最为敏感,但IIF法易受抗核抗体的影响,故不宜单独应用。MPO和PR3ELISA检测,对于原发性小血管炎的诊断特异性高,但由于需要价格昂贵的特异性纯化抗原,目前难以在我国普及推广。总抗原ELISA对于抗MPO和抗PR3抗体阳性血清检测的敏感性可达92%。结论:IIF法与MPO和PR3ELISA联合应用是较为理想的检测方案;IIF法与总抗原ELISA联合应用也具有较好的敏感性和特异性,且价格便宜,适于现阶段在我国推广应用。