蔬菜栽培土壤中沙门菌荧光定量PCR检测方法  

Quantitative Real-time PCR Detection Method of Salmonella in Vegetable Cultivated Soil

作　　者：吕新 刘兰英 李玥仁 LÜXin;LIU Lanying;LI Yueren(Institute of Quality Standards&Testing Technology for Agro-products,Fujian Academy of Agricultural Sciences,Fuzhou 350003;Fujian Key Laboratory of Agro-products Quality&Safety,Fuzhou 350003)

机构地区：[1]福建省农业科学院农业质量标准与检测技术研究所,福州350003 [2]福建省农产品质量安全重点实验室,福州350003

出　　处：《食品工业》2023年第7期321-325,共5页The Food Industry

基　　金：福建省科技计划公益类专项(2020R1022005);福建省农业科学院科技创新团队建设项目(CXTD2021011-3);福建省“5511”协同创新工程项目(XTCXGC2021020)。

摘　　要：以沙门菌特有侵袭蛋白A(invA)基因为靶基因,建立基于SYBR Green I嵌合荧光法的蔬菜栽培土壤中沙门菌实时荧光定量PCR(qPCR)检测方法。通过特异性试验和灵敏度试验检测,结果显示:该qPCR方法对沙门菌具有良好的特异性,其最低检出限为0.1 pg/μL的沙门菌DNA;所制作的qPCR扩增标准曲线在2~2×10^(5) CFU/g浓度之间有较好的线性关系,相关系数为0.998,可用于沙门菌的定量检测分析;该qPCR方法与国标方法对比分析蔬菜栽培土壤中沙门菌污染时具有相同的准确性,但检测时间从5~7 d缩短至6 h内,可满足蔬菜栽培土壤中沙门菌的定量检测需求。A pair of qPCR primers were designed based on the sequence of Salmonella specific invasion protein A(invA)gene,and a quantitative real-time PCR(qPCR)detection method for Salmonella in vegetable cultivated soil was established based on SYBR Green I chimeric fluorescence method.The results of the specificity test showed that the qPCR assay had good specificity;The results of the sensitivity test showed that the qPCR assay could detect Salmonella DNA at a minimum of 0.1 pg/μL;The standard curve of qPCR amplification revealed excellent linearity between 2 CFU/g and 2×10^(5) CFU/g concentration,and the PCR threshold cycle with correlation coefficient was 0.998,which could be used for quantitative analysis of Salmonella;Compared with the national standard method,the qPCR method had the same accuracy for Salmonella detection in vegetable cultivation soil,but the detection time was shortened from 5-7 d to 6 h,which could meet the needs of quantitative detection of Salmonella in vegetable cultivation soil.

关 键 词：实时荧光定量PCR 栽培土壤 沙门菌 

分 类 号：S63[农业科学—蔬菜学]