环介导等温核酸扩增检测缓冲液成分的探索

Study on the reaction system of loop-mediated isothermal amplification

目的探索环介导等温扩增(LAMP)检测反应体系缓冲液,为我国野战/灾害等紧急输血的血液筛查提供技术支持。方法通过对LAMP体系不同组分的梯度分析对检测结果的影响进行研究。结果扩增效率随着反应体系内Mg^(2+)浓度的增高而降低;LAMP反应的扩增效率随着反应体系内Betain浓度的增高而增高。随着Calcein浓度的降低,LAMP反应体系内的绿色荧光逐渐减弱。结论建立的反应体系具有较好的反应灵敏度及特异性,与成品化的试剂相似,可适用于临床血液病原体筛查。

Objective To explore loop-mediated isothermal amplification （LAMP） reaction system and provide technical support for field or disasters in emergency transfusion of blood screening. Methods The test results on the gradient LAMP different system components were anayzed. Results With the increasing of Mg^2＋ concentration, the efficiency of the amplification reaction system decreased; the amplification efficiency of the LAMP reaction increased with the increasing of the Betain concentration. With decreasing of Calcein concentration, green fluorescent in LAMP reaction system was gradually weakened. Conclusion The reaction system has good sensitivity and specificity which may be suitable for clinical blood screening.