Loop-mediated isothermal amplification for quick detection of Yersinia pestis

VernacularTitle:环介导恒温扩增技术快速检测鼠疫耶尔森菌
Author: Na FENG ; Yazhou ZHOU ; Yanxiao FAN ; Qiong WANG ; Yujing BI ; Yanping HAN ; Ruifu YANG ; Yusen ZHOU ; Xiaoyi WANG
Publication Type:Journal Article
Keywords: Yersinia pestis; loop-mediated isothermal amplification; detection
From: Military Medical Sciences 2015;(11):868-872
CountryChina
Language:Chinese
Abstract: Objective To establish a simple and quick loop-mediated isothermal amplification (LAMP)method for detection of Yersinia pestis.Methods LAMP Primers were designed based on the specific sequence 3a in Y.pestis chromosome.LAMP reaction results were detected using turbidity meter or visual method.The specificity of the constructed method was evaluated by detecting Y.pestis and its closely-related bacteria.The different dilution DNA template was detected with LAMP and PCR to evaluate the sensitivity of the method.Results Thirty strains of bacteria closely related to Y.pestis were detected by the constructed LAMP,and all the results were negative,indicating that the method had a very high specificity.The detection sensitivity of this LAMP assay was 20 pg of DNA per reaction,which was ten-fold that of the regular PCR.The detection reaction was completed in 25 min.Conclusion This LAMP method is quick,sensitive, specific and simple,which is expecked to become an effective method for rapid detection of Y.pestis on the scene.