基于双纳米金标记的侧流免疫层析法检测人骨钙素蛋白  

A Dual Gold Nanoparticle Conjugate-Based Lateral Flow Assay(LFA)Method for the Analysis of Human Serum Osteocalcin

作　　者：孙浩[1] 高向东[1] 吕炜锋[1] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009

出　　处：《药物生物技术》2017年第2期99-105,共7页Pharmaceutical Biotechnology

摘　　要：传统的基于纳米金的侧流免疫层析法灵敏度不高。为了克服这个缺点并且不增加额外的步骤,该研究通过双纳米金标记技术结合生物素-链霉亲和素蛋白放大系统,并利用金标定量阅读仪达到快速定量检测骨代谢标志物(以骨钙素为例)的目的。该试纸条的第一结合垫包被链霉亲和素-纳米金探针,第二结合垫包被抗骨钙素单抗Ab14318-纳米金-生物素化单链核苷酸探针,硝酸纤维素膜上的检测线包被抗骨钙素单抗Ab14319,质控线上包被羊抗鼠Ig G。通过对该侧流免疫层析试纸条与电化学免疫测定法(ECLIA)的相关性研究发现,两种方法有强相关性。该研究表明该试纸条拥有良好灵敏度、准确度和可靠性。与电化学免疫测定法相比,该方法更加简单、快速,并且不需要复杂的仪器设备,非常适合在检测现场或基层社区医院使用,应用前景非常广泛。For improving sensitivity without an additional operation step in a gold nanoparticle （AuNP）-based lateral flow assay（LFA） ,a novel,rapid and simple high-sensitive method utilizing the biotin-streptavidin binding system and double gold nanoparti-cles with different sizes as label was developed. AuNP-streptavidin conjugate was encapsulated in first conjugate pad;AuNP-anti-osteocalein antibody Ab14318 conjugate was encapsulated in second conjugate pad. Anti-osteocalcin antibody Ab14319 and goat anti-mouse IgG was dispensed in test line and control line, respectively, on the NC membrane. Nanogold quantitative immunoassay analy-zer was used to read the trips rapidly and quantitatively. Parallel analyses of human serum samples by gold nanoparticle conjugate-based LFA and elimmunoassay （ECLIA）obtained data in good agreement. And this Dual gold nanoparticleconjugate-based lateral flow assay had a good specificity, sensitivity and reliability. Compared with electrochemilnoassay, the gold nanoparticle conjugate-based LFA is simpler and more rapid method and it does not require complex instrumentsand equipment. It is very suitable for community hospital and near-patient testing.

关 键 词：骨钙素 纳米金 侧向免疫层析 生物素 链霉亲和素 

分 类 号：O657[理学—分析化学]