Pivovarov V D, Ryazantsev D Yu, Simonova M A, Yegorova T V, Khlgatian S V, Zavriev S K, Svirshchevskaya E V
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997 Russia.
Mechnikov Institute of Vaccines and Sera, Moscow, 105064 Russia.
Mol Biol (Mosk). 2018 Jul-Aug;52(4):727-734. doi: 10.1134/S0026898418040158.
Successful disease prevention and therapy critically depend on timely diagnosis of infections. Quantitative immuno-PCR (qiPCR) technology improves the sensitivity in the detection of antibodies to pathogens. A qiPCR-based assay was developed to determine IgG antibodies to Epstein-Barr virus (EBV) in the human blood serum. EBV nuclear protein 1 fragment (pEBV) was expressed in Escherichia coli. A synthetic single-stranded deoxyribonucleotide was conjugated to streptavidin, and the conjugate was used to detect рEBV-IgG1-biotin complexes by qiPCR. The IgG1 titers determined by qiPCR were compared to the results of enzyme-linked immunosorbent assay (ELISA). The sensitivity of qiPCR was one order of magnitude higher than that of ELISA. Thus, a highly sensitive qiPCR-based assay was developed to quantitate antibodies specific to the recombinant EBV antigen.
成功的疾病预防和治疗严重依赖于感染的及时诊断。定量免疫PCR（qiPCR）技术提高了病原体抗体检测的灵敏度。开发了一种基于qiPCR的检测方法来测定人血清中针对爱泼斯坦-巴尔病毒（EBV）的IgG抗体。EBV核蛋白1片段（pEBV）在大肠杆菌中表达。将合成的单链脱氧核糖核苷酸与链霉亲和素偶联，并用该偶联物通过qiPCR检测pEBV-IgG1-生物素复合物。将通过qiPCR测定的IgG1滴度与酶联免疫吸附测定（ELISA）的结果进行比较。qiPCR的灵敏度比ELISA高一个数量级。因此，开发了一种基于qiPCR的高灵敏度检测方法来定量重组EBV抗原特异性抗体。
