Poltavchenko Aleksandr Georgievich, Zaitsev Boris Nikolaevich, Ersh Anna Vasilievna, Korneev Denis Vladimirovich, Taranov Oleg Sviatoslavovich, Filatov Pavel Vladimirovich, Nechitaylo Oleg Viacheslavovich
a Laboratory of Methods of Immunochemical Diagnostic, State Research Center of Virology and Biotechnology "Vector" , Koltsovo , Novosibirsk , Russia.
b Laboratory of Ultrastructure Research, State Research Center of Virology and Biotechnology "Vector" , Koltsovo , Novosibirsk , Russia.
J Immunoassay Immunochem. 2016;37(5):540-54. doi: 10.1080/15321819.2016.1174134.
This study performed a comparative estimation of the detection systems with the use of conjugates based on peroxidase, alkaline phosphatase, colloidal gold, and the amplification system "Super-CARD" for multiplex dot-immunoassay of antibodies. The results of the study show that the sensitivity of the detection system with colloidal gold was approximately 8 times higher than that of the system with amplification "Super-CARD", 30 times higher than that of the system with conjugate of alkaline phosphatase, and 250 times higher than the sensitivity of the system with the peroxidase conjugate. Gold immunosols limit the direct detection of human IgG of 10 pg with dynamic range of optical signal change from 5 ng to 10 pg. This limit corresponds to a range of concentrations of IgG from 2.5 μg/mL to 5 ng/mL and covers the range of specific IgG concentrations, to which a typical natural immune response leads. The most typical reasons for aggregate formation during obtainment of colloidal gold and the binding of colloidal gold with biocomponents were explored. The method of minimization of particle clusters formation was suggested as well as the method for increasing stability of diluted preparations of probe by means of sedimentation of aggregates from the ready-made product.
本研究对基于过氧化物酶、碱性磷酸酶、胶体金的结合物检测系统以及用于抗体多重斑点免疫测定的“超级卡片”扩增系统进行了比较评估。研究结果表明，胶体金检测系统的灵敏度比“超级卡片”扩增系统高约8倍，比碱性磷酸酶结合物系统高30倍，比过氧化物酶结合物系统的灵敏度高250倍。金免疫溶胶可直接检测低至10 pg的人IgG，光信号变化的动态范围为5 ng至10 pg。此检测限对应于2.5 μg/mL至5 ng/mL的IgG浓度范围，涵盖了典型自然免疫反应所产生的特异性IgG浓度范围。探讨了在制备胶体金过程中形成聚集体以及胶体金与生物成分结合的最典型原因。提出了使颗粒聚集体形成最小化的方法，以及通过从成品中沉降聚集体来提高探针稀释制剂稳定性的方法。
