摘要
目的建立检测甲胎蛋白(AFP)的磁分离酶联免疫法(Magnetic affinity immunoassay,MAIA)。方法采用双抗体夹心法,用异硫氰酸荧光素(fluorescein isothiocyanate,FITC)和碱性磷酸酶(alkaline phosphatase,AP)分别标记识别不同表位的两种抗AFP单克隆抗体(mAb)4A3和5H7,以偶联羊抗FITC mAb包被的免疫磁珠作为固相载体,单磷酸酚酞(phenolphthalein monophosphate,PMP)做显色底物,建立检测AFP的MAIA法。结果成功实现了用MAIA法对AFP的定量检测,其检测灵敏度0.2 IU/ml,线性范围0.2-510 IU/ml,批内CV5.5%-9.3%,批间CV1.6%-9.7%。添加回收率101.4%-104.9%,稀释回收率78.3%-123%。与雅培AxSYM及i2 000的检测结果对比,相关系数分别为0.984和0.985。结论MAIA法的性能优于现有的ELISA和RIA试剂盒,接近进口同类试剂的水平,有助于为市场提供一种高质量而价廉的AFP测定试剂盒。
Objective To estabhsh MAIA for detecting AFP.Methods The inananoassay was based on antibody sandwich immtmoassay, two monoclonal antibodies（4A3 and 5H7） were used for conjugating with FITC and with alkaline phosphatase（AP） respectively, which incorporated magnetic solid phase separation. Magnetic beads were coupled with sheep anti-FITC antibody as solid pliase, and phenolphthalein monophosphate（PMP） was used as substrate to set up Magnetic Affinity/mmanoassay（MA/A） for detecting AFP. Results Established the MAIA for detcting AFP successfully. The sensitivity of the MAIA method was 0.2 IU/ml, the linear range was 0.2 - 510IU/ml, the intra-and inter-assay variation were 5.5 % - 9.3 % and 1.6 % - 9.7 % respectively, the addition and dilution recovery rate were in the range of 101.4% - 104.9% and 78.3% - 123% respectively. The correlation coefficient was 0.984 and 0.985 in comparison with Abbott AxSYM and i2 000 respectively. Conclusion AFP MAIA kit is better than AFP RIA and ELISA kit,which can provide a quality and cheaper AFP kit for market.
出处
《中国实验诊断学》
2008年第11期1427-1430,共4页
Chinese Journal of Laboratory Diagnosis
基金
国家自然基金资助项目(2006AA02090802)
关键词
甲胎蛋白
磁分离酶联免疫法
碱性磷酸酶
Alpha-fetoprotein
agnetic Affinity Immunoassay
alkaline phosphatase
