基于重组酶聚合酶扩增技术的日本血吸虫感染性钉螺检测体系的建立与应用

Establishment and Application of Detection System for Oncomelania Snails Infected by Schistosoma Japonicum Based on Recombinase Polymerase Amplification

确定日本血吸虫感染性钉螺是进行日本血吸虫病疫情预警的重要依据.现有日本血吸虫感染性钉螺的检测方法存在费时费力、设备昂贵、操作复杂等局限,因此研发一种易于推广和使用的日本血吸虫感染性钉螺检测方法十分必要.设计和开发一套基于重组酶聚合酶扩增(RPA)技术的日本血吸虫感染性钉螺检测体系.该检测体系可以在恒温(39℃)条件下、快速(20 min内)检测出浓度仅为10^(3)copies/μL的目标基因;结合简单的钉螺基因组DNA提取方法,该体系可以方便、快捷地检测出1/20的日本血吸虫感染性钉螺(即20只阴性钉螺中混有1只日本血吸虫感染性钉螺).

The determination of oncomelania snails infected by Schistosoma japonicum is an important basis for early warning of Schistosoma japonicum epidemic.The current methods for determining oncomelania snails infected by Schistosoma japonicum have some certain limitations,such as time-consuming and laborious,requiring expensive equipment,being complicated to operate.This study designes and develops a detection system for oncomelania snails infected by Schistosoma japonicum based on Recombinase Polymerase Amplification(RPA).The detection system established in this study can quickly(within 20 minutes)detect the target gene of in 10^(3)copies/μl under constant temperature(39℃).Combined with a simple DNA extraction method from oncomelania snail genome,this system can easily and quickly detect oncomelania snails infected by Schistosoma japonicum in 1/20(i.e.1 positive mixed in 20 oncomelania snails).