人甲胎蛋白时间分辨免疫荧光分析试剂盒的研制

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (03): 313-317.

人甲胎蛋白时间分辨免疫荧光分析试剂盒的研制

杭建峰¹, 吴英松¹, 余伟鸿², 徐伟文¹, 王从容³, 裘宇容³, 李明¹

1. 南方医科大学热带医学研究所, 广东, 广州, 510515;
2. 广州市达瑞抗体工程技术有限公司, 广东, 广州, 510663;
3. 南方医科大学南方医院检验科, 广东, 广州, 510515

出版日期:2005-03-20 发布日期:2005-03-20
基金资助:
收稿日期:2004-9-2。
基金项目:广州市研究开发基地建设项目(2003U130021)
作者简介:杭建峰(1976- ),男,在读硕士,电话:020-61648321,E-mail:hangjianfeng_ren@163.
通讯作者:李明(1959-),男,医学博士,教授

Preparation of time-resolved fluoroimmunoassay kit for human alpha-fetoprotein detection

HANG Jian-feng¹, WU Ying-song¹, YU Wei-hong², XU Wei-wen¹, WANG Cong-rong³, QIU Yu-rong³, LI Ming¹

1. 南方医科大学热带医学研究所, 广东, 广州, 510515;
2. 广州市达瑞抗体工程技术有限公司, 广东, 广州, 510663;
3. 南方医科大学南方医院检验科, 广东, 广州, 510515

Online:2005-03-20 Published:2005-03-20

摘要/Abstract

摘要： 目的研制人甲胎蛋白(hAFP)时间分辨免疫荧光分析(TRFIA)试剂盒。方法采用双抗体夹心法建立AFP TRFIA试剂盒,对试剂盒的各项指标进行评价。结果试剂盒的可测范围为1~1 000U/ml, 灵敏度为0.17U/ml,精密度良好,批内和批间的精密度分别为3.3%~5.9%,3.7%~ 6.5%。与CEA、CA12-5、CA19-9、CA15-3、白蛋白无交叉反应。稳定性试验表明试剂可以在4℃稳定1年,37℃稳定7d。426份正常血清标本测试该试剂盒的正常参考值范围是0~12U/ml。用本试剂盒与国外同类试剂盒同时检测60份血清标本,其相关系数为0.995。结论试剂盒各项指标(灵敏度、精密度、特异性、稳定性、准确度)均达到临床检测要求,可替代国外同类产品试剂盒。

Abstract: Objective To prepare time-resolved fluoroimmunoassay (TRFIA) kit for detecting human alpha-fetoprotein (hAFP).Methods Sandwich TRFIA kit for hAFP detection was developed using monoclonal antibody of anti-AFP.Results AFP-TRFIA kit was capable of detecting AFP within the range of 1-1 000 U/ml with sensitivity of 0.17 U/ml and without cross-reactivity with CEA, CA12-5, CA15-3, or CA19-9. The intra- and inter-assay coefficients of variation were (3.3-5.9)% and (3.7-6.5)%, respectively. The prepared AFP-TRFIA reagent could withstand preservation at 4℃ for 1 year and at 37℃ for 7 days with the cutoff value of 12U/ml in healthy subjects (n=426). The correlation coefficient of the detection results between this kit and commercially available AFP kit (Wallac OY, Finland) for 60 blood samples was 0.995.Conclusion The prepared TRFIA kit for hAFP detection meets the demand of clinical application with good sensitivity, precision, specificity, stability and accuracy.

中图分类号:

R392-33

杭建峰¹, 吴英松¹, 余伟鸿², 徐伟文¹, 王从容³, 裘宇容³, 李明¹. 人甲胎蛋白时间分辨免疫荧光分析试剂盒的研制[J]. 南方医科大学学报, 2005, 25(03): 313-317.

HANG Jian-feng¹, WU Ying-song¹, YU Wei-hong², XU Wei-wen¹, WANG Cong-rong³, QIU Yu-rong³, LI Ming¹. Preparation of time-resolved fluoroimmunoassay kit for human alpha-fetoprotein detection[J]. Journal of Southern Medical University, 2005, 25(03): 313-317.

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