Bodulev Oleg L, Gribas Anastasia V, Sakharov Ivan Yu
Department of Chemistry, Lomonosov Moscow State University, Leninskie gory, Moscow 119991, Russia.
Department of Chemistry, Lomonosov Moscow State University, Leninskie gory, Moscow 119991, Russia.
Anal Biochem. 2018 Feb 15;543:33-36. doi: 10.1016/j.ab.2017.11.026. Epub 2017 Dec 5.
A sensitive sandwich assay for hepatitis B virus (HBV) DNA detection based on use of commercial CL-ELISA microplates was developed. To reveal the target the covalent conjugate of reporter oligonucleotide and horseradish peroxidase (HRP) was synthesized. An employment of enhanced chemiluminescence reaction, where 3-(10'-phenothiazinyl)propionic acid/N-morpholinopyridine pair was used as enhancer of HRP-catalyzed chemiluminescence, permitted to measure the enzyme activity of the conjugate with high sensitivity. Under the favorable conditions the limit of detection and a linear range of the assay were 3 pM and 0.07-2.0 nM, respectively. The coefficient of variation (CV) for determination of HBV DNA concentrations within the working range was lower than 4%. The obtained results demonstrated that the developed assay had high sensitivity and precision.
基于商用化学发光酶联免疫吸附分析（CL-ELISA）微孔板开发了一种用于检测乙型肝炎病毒（HBV）DNA的灵敏夹心分析方法。为了揭示目标物，合成了报告寡核苷酸与辣根过氧化物酶（HRP）的共价缀合物。采用增强化学发光反应，其中3-（10'-吩噻嗪基）丙酸/N-吗啉吡啶对用作HRP催化化学发光的增强剂，从而能够高灵敏度地测量缀合物的酶活性。在有利条件下，该分析方法的检测限和线性范围分别为3 pM和0.07-2.0 nM。工作范围内测定HBV DNA浓度的变异系数（CV）低于4%。所得结果表明，所开发的分析方法具有高灵敏度和精密度。
