食品中亚利桑那菌环介导等温扩增检测方法的建立

Detection of Salmonella arizonae in food by loop-mediated isothermal amplification

应用环介导等温扩增(loop mediated isothermal amplification,LAMP)技术,研究建立一种快速、敏感、特异的检测亚利桑那菌(Salmonella arizonae)方法,为食品安全检测工作中亚利桑那菌检测提供一种快速准确工具。针对亚利桑那菌spv A基因中39个保守且特异的碱基序列,设计LAMP特异性引物,利用实时浊度仪对引物及反应条件进行优化,并对检测方法的灵敏度和特异性进行验证。该方法在63℃恒温下作用40 min,亚利桑那菌DNA获得高效率的特异性扩增;其检出限量为培养菌液浓度10~6CFU/m L,敏感性高;通过对25份沙门氏菌阳性食品的LAMP检测,检出2例亚利桑那菌。该文所建立的亚利桑那菌环介导等温扩增LAMP检测方法具有特异、灵敏、简便快捷等特点,适用于食品安全检测中亚利桑那菌的快速筛检。

The objective of the study is to establish a rapid detection of Salmonella arizonae by a loop-mediated isothermal amplification assay( LAMP). The primers were designed according to the spv A gene sequence and sensitivity,and specificity of the LAMP detection method was analyzed by real-time turbidity.The method of LAMP shows a highly efficient amplification for Salmonella arizonae target gene which performed at 63 ℃ for 40 min. The detection limit was 106CFU/m L of bacillus Concentration. 25 Salmonella positive food samples were detected by LAMP and 2 samples were identified Salmonella arizonae.The results suggested that LAMP method is effective,specific and sensitive,and it is suitable for the detection of Salmonella arizonae in food.