Recombinase polymerase amplification (RPA) is a technique that is used to specifically amplify a target nucleic acid sequence. Unlike the polymerase chain reaction (PCR), RPA is performed at a constant temperature between 37 and 42°C. Therefore, it can be potentially used for the onsite detection of various pathogens when combined with DNA extraction and amplicon detection techniques. In this study, we prepared recombinant recombinase and single-stranded DNA-binding protein from T4 phage and used them to examine the effects of reaction conditions and additives on the efficiency of RPA. The results revealed that the optimal pH was 7.5–8.0, optimal potassium acetate concentration was 40–80 mM, and optimal reaction temperature was 37–45°C although dimethyl sulfoxide at 5% v/v and formamide at 5% v/v inhibited the reaction. Our results suggest that RPA could be conducted using a wider range of optimal reaction conditions than those required for PCR and that RPA is highly suitable for point-of-care use.

重组酶聚合酶扩增（RPA）是用于特异性扩增靶核酸序列的技术。与聚合酶链反应（PCR）不同，RPA在37至42°C的恒定温度下进行。因此，与DNA提取和扩增子检测技术结合使用时，可潜在地用于各种病原体的现场检测。在这项研究中，我们从T4噬菌体中制备了重组重组酶和单链DNA结合蛋白，并用它们来检验反应条件和添加剂对RPA效率的影响。结果显示，尽管5％v / v的二甲亚砜和5％v / v的甲酰胺均被抑制，但最佳pH值为7.5-8.0，最佳乙酸钾浓度为40-80 mM，最佳反应温度为37-45°C。反应。