Polyclonal antibodies against potato spindle tuber viroid RNA.

Yuri Drygin, Tatiana Gasanova, Konstantin Butenko
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引用次数: 1

Abstract

Ribonucleic acid (RNA) can act as a hapten in the direct immunization of animals. For antigen synthesis, 65 mg of viroid RNA were obtained by in vitro transcription of the recombinant DNA. We received a reasonable immune response in mice and rabbits with synthesized conjugate viroid RNA-lysozyme. Analyses of polyclonal mouse and rabbit antisera as well as estimates of antibody specificity were performed by dot-Enzyme Linked Immunosorbent Assay (ELISA), sandwich ELISA, and northern immunoblotting. Antiserum obtained showed strong cross-reactions with cellular RNA. The viroid polyclonal antibody cross-reactions with cellular RNAs were depleted via titration antibodies by the plant cellular or commercial yeast RNA. We successfully used antibodies against the viroid RNA-lysozyme antigen to detect the wild-type potato viroid and diagnose potato viroid infection. We presume that intrinsic cross-reactions of RNA antibodies are potentially dangerous after nucleic acid vaccination. Research into the specificity of antibodies against viral RNAs is underway.

马铃薯梭形块茎类病毒RNA多克隆抗体。
核糖核酸(RNA)在动物直接免疫中可作为半抗原。在抗原合成方面,通过体外转录重组DNA获得65 mg的类病毒RNA。合成的共轭类病毒rna -溶菌酶在小鼠和家兔中获得了合理的免疫应答。采用点酶联免疫吸附试验(ELISA)、夹心酶联免疫吸附试验(sandwich ELISA)和northern免疫印迹法对多克隆小鼠和兔抗血清进行分析,并估计抗体特异性。所得抗血清与细胞RNA有较强的交叉反应。与细胞RNA交叉反应的类病毒多克隆抗体通过植物细胞或商业酵母RNA的滴定抗体被耗尽。利用类病毒rna溶菌酶抗原抗体成功检测了野生型马铃薯类病毒,并对马铃薯类病毒感染进行了诊断。我们推测核酸接种后RNA抗体的内在交叉反应具有潜在的危险性。针对病毒rna的抗体特异性的研究正在进行中。
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