[1]赵 萍,陈 飞,周家宏.花生制品中金黄色葡萄球菌SYBR Green qPCR快检方法的评价研究[J].南京师范大学学报(工程技术版),2023,23(02):069-76.[doi:10.3969/j.issn.1672-1292.2023.02.010]
　Zhao Ping,Chen Fei,Zhou Jiahong.The Evaluation Study of SYBR Green qPCR Rapid Detection Method for Staphylococcus aureus in Peanut Products[J].Journal of Nanjing Normal University(Engineering and Technology),2023,23(02):069-76.[doi:10.3969/j.issn.1672-1292.2023.02.010]
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花生制品中金黄色葡萄球菌SYBR Green qPCR快检方法的评价研究
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南京师范大学学报（工程技术版）[ISSN:1006-6977/CN:61-1281/TN]

卷:
23卷
期数:
2023年02期
页码:
069-76
栏目:
食品科学与工程
出版日期:
2023-06-15

文章信息/Info

Title:
The Evaluation Study of SYBR Green qPCR Rapid Detection Method for Staphylococcus aureus in Peanut Products
文章编号:
1672-1292(2023)02-0069-08
作者:
赵 萍1; 2; 陈 飞2; 3; 周家宏3
(1.南京师范大学食品与制药工程学院,江苏 南京 210023)
(2.南京师范大学南京市特色生物资源功能成分开发工程研究中心,江苏 南京 210023)
(3.南京师范大学生命科学学院,江苏 南京 210023)
Author(s):
Zhao Ping1; 2; Chen Fei2; 3; Zhou Jiahong3
(1.School of Food Science and Pharmaceutical Engineering,Nanjing Normal University,Nanjing 210023,China)
(2.Nanjing Engineering Research Center for Functional Components Development of Featured Biological Resources,Nanjing Normal University,Nanjing 210023,China)
(3.School of Life Sciences,Nanjing Normal University,Nanjing 210023,China)
关键词:
金黄色葡萄球菌; 花生制品; SYBR Green实时荧光定量PCR; 快速检测; 评价研究
Keywords:
staphylococcus aureus; peanut product; SYBR Green quantitative real-time PCR(qPCR); rapid detection; evaluation study
分类号:
TS207.4
DOI:
10.3969/j.issn.1672-1292.2023.02.010
文献标志码:
A
摘要:
金黄色葡萄球菌是花生及其制品中常见的一种食源性致病菌,针对金黄色葡萄球菌的快速检测方法对实现花生及其制品中的食品安全防控有重要意义. 根据金黄色葡萄球菌的耐热核酸酶基因(nuc)设计了3对特异性引物,同时优化了退火温度,成功建立了一种特异性引物的SYBR Green实时荧光定量PCR(qPCR)快速检测方法,进一步对该方法的特异性、重现性及灵敏度进行了验证. 以8种模拟污染花生加工产品为实验对象,对该方法与荧光定量PCR试剂盒-探针法及商品化Baird-Parker干粉培养基法进行了评价研究. 结果表明,建立的SYBR Green qPCR快速检测方法特异性和重现性良好,最低检测限为8.99 copies/μL,其灵敏度比荧光定量PCR试剂盒-探针法高2个数量级. SYBR Green qPCR快速检测方法具有成本低、检测快速、特异性好、灵敏度高的优点,可用于花生及其制品中金黄色葡萄球菌的快速检测,也可为防控其他食品中的金黄色葡萄球菌污染提供参考.
Abstract:
Staphylococcus aureus is a common causative pathogen in peanuts and peanut products. Rapid detection of S.aureus is of great significance for the prevention and control of microbial contamination of peanut products. In this study,we design three pairs of specific primers according to the nuc gene of S.aureus,and a SYBR Green qPCR rapid detection method is established by optimizing the annealing temperature. The specificity,repeatability and sensitivity of the method are further evaluated. Finally,eight simulated contaminated peanut products are used as experimental objects to evaluate SYBR Green qPCR method,fluorescence quantitative PCR kit-probe method and commercial Baird-Parker dry powder medium method. The results show that the established method can specificity and repeatability detect S.aureus,the minimum detection limit is 8.99 copies/μL,and its sensitivity is two orders of magnitude higher than that of probe PCR kit method. The SYBR Green qPCR method has the advantage of economy,rapidity,specificity and high sensitivity,is suitable for the rapid detection of S.aureus in peanuts and peanut products,and can also provide reference for the prevention and control of S.aureus contamination in other foods.

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备注/Memo

备注/Memo:
收稿日期:2021-11-21.
基金项目:“十三五”国家重点研发计划重点专项项目(2018YFC1603400).
通讯作者:陈飞,博士,副教授,研究方向:食品生物技术. E-mail:chenfei@njnu.edu.cn
更新日期/Last Update: 2023-06-15