摘要
本文以羧基96孔板为分离载体,核酸适配体作为分子特异性识别元件,聚苯乙烯微球作为放大载体,辣根过氧化物酶为标记物,构建了化学发光(CL)高灵敏度凝血酶检测新技术.实验结果表明:该放大技术不但灵敏度高,且抗干扰能力强,其他蛋白质如IgG、IgM、IgA、IgE、IFN均无明显干扰.聚苯乙烯微球放大体系中凝血酶的线性范围为7.8～250pmol/L,最低检测浓度可达3.9pmol/L;而不放大检测技术的线性范围为0.94～30nmol/L,最低检测浓度为0.46nmol/L,放大体系将检测灵敏度提高100多倍.综合而言,基于适配体识别和聚苯乙烯微球放大的凝血酶CL检测新技术具有通量大、简单快速和灵敏度高的特点,有望在凝血酶高通量检测领域获得应用.
A novel chemiluminescence（CL） immunoassay for the amplified determination of thrombin is proposed,by using two aptamer sequences as recognition elements,96-well microplate as a reaction carrier,polystyrene microspheres as an amplification platform and horseradish peroxidase（HRP） as a CL label.The results show that the amplified CL technique couples the high sensitivity of CL analysis with effective discrimination against unwanted proteins such as IgG,IgM,IgA,IgE and IFN.An excellent linearity from 7.8 to 250 pM with the lowest detection limit of 3.9 pM was reached for the amplified technique whereas a good linear correlation from 0.98 to 30 nM with a detection limit of 0.46 nM was achieved for the nonamplified method,which was more than 100-fold less sensitive than using the amplified approach.Overall,this proposed detection scheme is time-effective with high sensitivity and throughout.These features,as well as its operation convenience,make it a promising alternative to thrombin detection methods.
出处
《中国科学：化学》
CAS
CSCD
北大核心
2011年第10期1622-1628,共7页
SCIENTIA SINICA Chimica
基金
重大新药创制专项(2009ZX09301-011)
国家重大基础研究计划(2007CB935800)
国家自然科学基金(20975026)
教育部博士点基金(20090071110056)资助
