|Table of Contents|

The Evaluation Study of SYBR Green qPCR Rapid Detection Method for Staphylococcus aureus in Peanut Products(PDF)

南京师范大学学报（工程技术版）[ISSN:1006-6977/CN:61-1281/TN]

Issue:
2023年02期
Page:
69-76
Research Field:
食品科学与工程
Publishing date:

Info

Title:
The Evaluation Study of SYBR Green qPCR Rapid Detection Method for Staphylococcus aureus in Peanut Products
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)
Keywords:
staphylococcus aureus; peanut product; SYBR Green quantitative real-time PCR(qPCR); rapid detection; evaluation study
PACS:
TS207.4
DOI:
10.3969/j.issn.1672-1292.2023.02.010
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.

References:

[1]惠媛媛,彭海帅,王毕妮,等. 基于适配体的光学和电化学法对食源性致病菌检测的研究进展[J]. 中国农业科学,2021,54(11):2419-2433.
[2]李莹. 中国食品安全及其监管制度研究[D]. 长春:吉林大学,2014.
[3]张宁,王建华. 食品微生物标准和食品安全[J]. 品牌与标准化,2014(8):23.
[4]VAIYAPURI M,JOSEPH T C,RAO B M,et al. Methicillin-resistant Staphylococcus aureus in seafood:Prevalence,laboratory detection,clonal nature,and control in seafood chain[J]. Journal of Food Science,2016,84(12):3341-3351.
[5]DA SILVA A C,RODRIGUES M X,SILVA N C C. Methicillin-resistant Staphylococcus aureus in food and the prevalence in Brazil:a review[J]. Brazilian Journal of Microbiology,2020,51(1):347-356.
[6]RUBAB M,SHAHBAZ H M,OLAIMAT A N,et al. Biosensors for rapid and sensitive detection of Staphylococcus aureus in food[J]. Biosensors & Bioelectronics,2018,105:49-57.
[7]SHIMAA E,MOHAMMED Z. Ultrasensitive peptide-based multiplexed electrochemical biosensor for the simultaneous detection of Listeria monocytogenes and Staphylococcus aureus[J]. Mikrochimica Acta:An International Journal of Physical and Chemical Methods of Analysis,2020,187(9):486.
[8]李传友,曾宝锋,赵燕英,等. 恒温隔绝式聚合酶链式反应检测食品中金黄色葡萄球菌[J]. 食品科学,2022,43(2):339-345.
[9]宋明辉,施春雷,李琼琼,等. 速冻面米制品中金黄色葡萄球菌和肠毒素A基因三重PMA-qPCR快检方法的建立[J]. 中国食品学报,2020,20(7):244-252.
[10]PAPADOPOULOS P,ANGELIDIS A S,PAPADOPOULOS T,et al. Staphylococcus aureus and methicillin-resistant S.aureus(MRSA)in bulk tank milk,livestock and dairy-farm personnel in north-central and north-eastern Greece:Prevalence,characterization and genetic relatedness[J]. Food Microbiology,2019,84:103249.
[11]国家卫生和计划生育委员会,国家食品药品监督管理总局. 食品安全国家标准 食品微生物学检验 金黄色葡萄球菌检验:GB 4789.10—2016[S]. 北京:中国标准出版社,2016.
[12]LEE W,PARK Y,SHRIVASTAVA S,et al. A fully integrated bacterial pathogen detection system based on count-on-a-cartridge platform for rapid,ultrasensitive,highly accurate and culture-free assay[J]. Biosensors and Bioelectronics,2020,152:112007.
[13]王海军,马福敏,张革新,等. 快速检测在食品安全监管中应用的研究[J]. 食品研究与开发,2013,34(19):118-121.
[14]汪华,孔英戈,潘萍,等. 快速检测技术在食用农产品安全监管中的应用[J]. 食品安全导刊,2016(33):12-13.
[15]李菊,谢建军,王璐,等. 基于质量评价的孔雀石绿快检试剂盒筛选策略[J]. 食品安全质量检测学报,2020,11(3):875-882.
[16]YANG Y,SUN X D,YUAN Y W,et al. Detection of Staphylococcus aureus in dairy products by polymerase chain reaction assay[J]. Agricultural Sciences in China,2007,6(7):857-862.
[17]廖光华,代佳君,邓钰蓱,等. 基于nuc基因的奶牛乳房炎性金黄色葡萄球菌PCR检测方法的建立[J]. 塔里木大学学报,2019,31(4):7-11.
[18]郭慧芳,李宁,王白玉,等. 鸡Caspase-1基因SYBR GreenⅠ荧光定量PCR检测方法的建立[J]. 中国预防兽医学报,2021,43(5):521-526.
[19]王礼. 食品中金黄色葡萄球菌的检测及防控技术[J]. 中国食品,2021(17):142-143.
[20]LOIR Y L,BARON F,GAUTIER M. Staphylococcus aureus and food poisoning[J]. Genetics & Molecular Research,2003,2(1):63-76.
[21]SHEET O H,GRABOWSKI N T,KLEIN G,et al. Development and validation of a loop mediated isothermal amplification(LAMP)assay for the detection of Staphylococcus aureus in bovine mastitis milk samples[J]. Molecular and Cellular Probes,2016,30(5):320-325.
[22]后来旺,李达容,邓波,等. 金黄色葡萄球菌RAA-LFD快速检测方法的建立与应用[J]. 食品科学,2022,43(4):331-339.
[23]郭建平,万佳佳,陆兆新,等. 基于可视化环介导等温扩增技术快速检测金黄色葡萄球菌[J]. 食品科学,2019,40(20):325-331.
[24]母润红,聂丹丹,赵明,等. 微滴式数字聚合酶链反应检测金黄色葡萄球菌和蜡样芽孢杆菌[J]. 吉林医药学院学报,2021,42(2):81-83.
[25]孙冰清,姜芹,张文刚,等. 基于qPCR检测金黄色葡萄球菌3种肠毒素基因的方法研究[J]. 食品安全质量检测学报,2020,11(9):2798-2805.

Memo

Memo:
-
Last Update: 2023-06-15