应用多重实时荧光PCR检测蜡样芽胞杆菌毒力基因

Development of a mutiplex real-time PCR assay for detection of the toxin gene to Bacillus cereus

摘要: 目的:建立一种快速、准确、简便的蜡样芽胞杆菌毒力基因检测方法,为蜡样芽胞杆菌更深入的研究提供依据和便利。方法:采用TaqMan探针法设计了三重hblA,hblC,hblD基因,三重nheA,nheB,nheC基因,三重ces,entFM,bceT基因的多重实时荧光PCR体系检测蜡样芽胞杆菌毒力基因。结果:多重体系对蜡样芽胞杆菌毒力基因的检测具有良好的特异性和灵敏性,其最低检出限为63 CFU/mL。结论:利用上述体系对蜡样芽胞杆菌的毒力基因进行检测能够省时省力,特异性、灵敏性良好,具有实际应用价值。

Abstract: Objective:To develop a fast,accurate and simple method to detect the toxin genes in Bacillus cereus,and provide basis and convience for a deeper research. Methods:Three multiplex real-time fluorescence PCR systems were developed by TaqMan Probe method for the detection of toxin genes of hblA-hblC-hblD for the first PCR reaction system,nheA-nheB-nheC for the second,entFM-bceT-ces for the third. Results:The detection results of toxin genes in Bacillus cereus indicated excellent specificity and sensitivity,detection of limit of bacterial concentration was 63 CFU/mL. Conclution:The systems above had the characteristics of time saving and labor saving. They had excellent specificity and sensitivity.