斑点免疫金渗滤法定量测定血清淀粉样蛋白A方法的建立及性能评估

doi:10.3969/j.issn.1673-8640.2013.09.024

检验医学 ›› 2013, Vol. 28 ›› Issue (9): 835-840.DOI: 10.3969/j.issn.1673-8640.2013.09.024

• 技术研究与评价.论著 • 上一篇下一篇

斑点免疫金渗滤法定量测定血清淀粉样蛋白A方法的建立及性能评估

陈长强^¹,杨晶^²,顾志冬^¹,万颖蕾^¹,毛客自^¹,冯晓静^¹,徐建新^²,樊绮诗^¹

1.上海交通大学医学院附属瑞金医院检验科,上海 200025;
2.上海奥普生物医药有限公司研发中心,上海 201201

收稿日期:2013-09-25 修回日期:2013-09-25 出版日期:2013-09-15 发布日期:2013-09-25
通讯作者: 樊绮诗,联系电话:021-67888236。
作者简介:陈长强,男,1986年生,硕士,技师,主要从事临床检验工作。

Establishment and performance evaluations of dot immuno-gold filtration assay for the quantitative determination of serum amyloid A

CHEN Changqiang^¹,YANG Jing^²,GU Zhidong^¹,WAN Yinglei^¹,MAO Kezi^¹,FENG Xiaojing^¹,XU Jianxin^²,FAN Qishi^¹.

1.Department of Clinical Laboratory, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China;
2.Shanghai Upper Bio-Tech Pharma Co.,Ltd., Shanghai 201201, China

Received:2013-09-25 Revised:2013-09-25 Online:2013-09-15 Published:2013-09-25

摘要/Abstract

摘要：

目的建立斑点免疫金渗滤法(DIGFA)定量测定血清淀粉样蛋白A(SAA)并评估其分析性能。方法选用2株商品化抗SAA单克隆抗体,分别结合于硝酸纤维素膜和胶体金,制备免疫金渗滤试剂板条(SAA-DOT)以定量检测SAA。按照EP文件,评估SAA-DOT的分析性能(检测限、精密度、准确性、线性范围等)。结果 SAA-DOT最低检测限为5 mg/L;10和100 mg/L的批内变异系数(CV)分别为9.07%和10.21%,天间CV分别为11.33%和11.53%;线性范围为5～230 mg/L;SAA-DOT的测定结果与商品化乳胶增强速率散射比浊法(LERN)试剂具有良好的相关性(r=0.995,P<0.05)。表观健康者血清SAA浓度中位数为5.7 mg/L,95%可信区间为0~9.6 mg/L。结论基于DIGFA的SAA-DOT简便、快速,能在5 min内完成检测,各项分析性能指标达到国家食品药品监督管理局(SFDA)对体外诊断试剂的要求,可用于临床患者血清标本的检测。

关键词: 斑点免疫金渗滤法, 血清淀粉样蛋白A, 即时检验, 乳胶增强速率散射比浊法

Abstract:

Objective To establish a dot immuno-gold filtration assay (DIGFA) for the quantitative determination of serum amyloid A (SAA),and evaluate its analysis performance. Methods Serum DIGFA-based SAA detection reagent (SAA-DOT) was established using 2 anti-SAA monoclonal antibodies being coated with nitrocellulose filter and labeled by colloidal gold, respectively. According to EP files, the SAA-DOT analysis performances (such as detection limit,precision,accuracy,linear range,et al) were evaluated. Results The detection limit of SAA-DOT was 5mg/L. The within-run coefficients of variation (CV) were 9.07% and 10.21% respectively, whereas the inter-day CV were 11.33% and 11.53% respectively, when serum SAA concentrations were 10 and 100 mg/L. The detection linear range was from 5 mg/L to 230 mg/L. SAA-DOT had a good correlationship with latex-enhanced rate nephelometry (LERN) reagent (r=0.995,P<0.05). The median of serum SAA was 5.7 mg/L, and the 95% confidence interval was 0-9.6 mg/L in healthy subjects. Conclusions The DIGFA-based SAA-DOT is convenient, and the reaction can be finished within 5 min. Its performance meets the State Food and Drug Administrration (SFDA) requirements for in vitro diagnostic reagents,which is suitable for the clinical applifcation of serum detection.

Key words: Dot immuno-gold filtration assay, Serum amyloid A, Point-of-care test, Latex-enhanced rate nephelometry

中图分类号:

R446.61

陈长强,杨晶,顾志冬,万颖蕾,毛客自,冯晓静,徐建新,樊绮诗. 斑点免疫金渗滤法定量测定血清淀粉样蛋白A方法的建立及性能评估[J]. 检验医学, 2013, 28(9): 835-840.

CHEN Changqiang,YANG Jing,GU Zhidong,WAN Yinglei,MAO Kezi,FENG Xiaojing,XU Jianxin,FAN Qishi.. Establishment and performance evaluations of dot immuno-gold filtration assay for the quantitative determination of serum amyloid A[J]. , 2013, 28(9): 835-840.

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斑点免疫金渗滤法定量测定血清淀粉样蛋白A方法的建立及性能评估

Establishment and performance evaluations of dot immuno-gold filtration assay for the quantitative determination of serum amyloid A

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