摘要
为建立检测沙门氏菌的快速、特异、灵敏的荧光定量PCR方法,以沙门氏菌inv A基因为目的基因设计引物,并进行特异性、灵敏度和重复性实验。结果显示,建立的SYBR GreenⅠ荧光定量PCR检测沙门氏菌的方法具有极强的特异性,其中沙门氏菌检出限为86copies/m L,标准曲线的相关系数为0.999,扩增效率为104%,不同浓度质粒重复性扩增实验Ct值的变异系数均<3%,符合WHO对中等实验室提出的标准(CV 3%),显示了良好的重复性。结果表明该方法具有快速、简单、灵敏度高、特异性强等优点,可应用于食品卫生监管、商品检验检疫以及临床诊断等领域。
According to the invA gene of Salmonella spp.,a pair of primers was designed to establish a real-time PCR.The rapid, specific, sensitive detection of Salmonella spp. was achieved, and the evaluation of detection sensitivity,specificity and repeatability was carried out.This method had good specificity.The limit of detection of the method was 86copies,correlation coefficient of the standard curve was 0.999 and amplification efficiency was 104%.Different concentrations of plasmid repetitive amplification coefficients of variation were less 3% ,accord with standard preferred for moderately laboratory by WHO, indicating a good reliability.The SYBR Green I real-time PCR detection technology was rapid,simple, specific,and sensitive,which may be widely applied in the detection of Salmonella spp. in the field of food sanitation supervision, commodity inspection and detection, clinical diagnosis, etc.
出处
《食品工业科技》
CAS
CSCD
北大核心
2015年第3期147-150,共4页
Science and Technology of Food Industry
基金
国家“十二五”支撑计划(2012BAD12B06)
国家星火计划项目(2012GA600001)
