PDF(Pubmed)
Abstract：
Recombinase polymerase amplification (RPA) is an isothermal in vitro nucleic acid amplification technique that has been adopted for simple, robust, rapid, reliable diagnostics of nematodes. In this study, the real-time RPA assay and RPA assay combined with lateral flow dipsticks (LF-RPA) have been developed targeting the ITS rRNA gene of the British root-knot nematode, Meloidogyne artiellia. The assay provided specific and rapid detection of this root-knot nematode species from crude nematode extracts without a DNA extraction step with a sensitivity of 0.125 second-stage juvenile (J2) specimen per a reaction tube for real-time RPA during 11 min and a sensitivity of 0.5 J2 specimens per a reaction tube for LF-RPA during 25 min. The RPA assays were validated with a wide range of non-target root-knot nematodes. The LF-RPA assay has great potential for nematode diagnostics in the laboratory having minimal available equipment.
摘要：
重组酶聚合酶扩增(RPA)是一种等温的体外核酸扩增技术,健壮,快速,可靠的线虫诊断。在这项研究中,已针对英国根结线虫的ITSrRNA基因开发了实时RPA测定和RPA测定结合侧流试纸（LF-RPA），南方根结线虫。该测定法提供了从粗线虫提取物中对这种根结线虫物种的特异性和快速检测，而无需进行DNA提取步骤，每个反应管对11分钟内实时RPA的灵敏度为0.125第二阶段少年（J2）样本，并且在25分钟内每个反应管对LF-RPA的灵敏度为0.5J2样本。用多种非目标根结线虫验证了RPA测定。LF-RPA测定在具有最少可用设备的实验室中具有用于线虫诊断的巨大潜力。
