摘要
研制一种低成本、快速、准确的三抗体夹心酶联免疫(Three antibodies sandwich ELISA,TAS-ELISA)检测试剂盒。采用戊二醛法、改良过碘酸钠法分别制备碱性磷酸酶(Alkaline Phosphatase,AP)、辣根过氧化物酶(Horseradish Peroxidase,HRP)标记的山羊抗兔IgG,比较单核细胞增生性李斯特菌(Listeria monocytogenes,LM)TAS-ELISA试剂盒两种酶标抗体的检测效果,并考察了试剂盒的检测灵敏度、特异性、稳定性、保存周期等指标。结果表明,采用改良过碘酸钠法制备的HRP-IgG效果好,克分子比最高可达2.9,酶结合率达27%;试剂盒检测周期6h,检测成本为3.5元/孔,特异性实验、干扰实验、重复性实验、稳定性实验表明采用该方法制备的酶标抗体,特异性好、结果稳定可靠。自制TAS-ELISA试剂盒检测性能可达到商业化试剂盒水平,为该产品的产业化提供了一定的理论依据。
To develop a low-cost,rapid and accurate TAS-ELISA KIT.This study preparated AP-IgG and HRP-IgG by glutaraldehyde method and improved periodate method respectively and compared two enzyme conjugates of Listeria monocytogenes TAS-ELISA KIT,further investigated some indexes including the sensitivity,specificity,stability,shelf life.The results showed that HRP-IgG preparated with improved periodate method was more effective,the molar ratios was 2.9,the rate of enzyme-labeled was 27%.The detection time of KIT was 6h,the detection cost was $3.5/well.Specific experiment,interference experiment,repetitive experiment and stable experiment indicated that the HRP-IgG preparated with improved periodate method was specific and reliable.The performance of self-made TAS-ELISA KIT could reach commercial KIT,it also could provide theoretical basis for industrialization of the product.
出处
《食品工业科技》
CAS
CSCD
北大核心
2012年第15期301-306,310,共7页
Science and Technology of Food Industry
基金
质检总局科研项目(GSCIQ_2010IK220)
