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  被引量:7

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

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作  者:陈长强[1] 杨晶 顾志冬[1] 万颖蕾[1] 毛客自[1] 冯晓静[1] 徐建新 樊绮诗[1] 

机构地区:[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

出  处:《检验医学》2013年第9期835-840,共6页Laboratory Medicine

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(SAADOT)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。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

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 

分 类 号:R446.61[医药卫生—诊断学]

 

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