Food and Fermentation Industries >

2023 , Vol. 49 >Issue 22: 279 - 286

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.035031

Simultaneous detection of Listeria monocytogenes and Staphylococcus aureus based on dual DARQ-LAMP method

  • CHEN Qingying ,
  • LIU Dan ,
  • ZHANG Xingguo ,
  • WEI Jinyuan ,
  • ZHONG Qingping
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  • (Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University, Guangzhou 510642, China)

Received date: 2023-02-06

  Revised date: 2023-02-20

  Online published: 2023-12-25

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Abstract

Loop-mediated isothermal amplification (LAMP) is an effective method for rapid detection of foodborne pathogens. In this study, DARQ-LAMP (detection of amplification by release of quenching and loop-mediated isothermal amplification) method was established for simultaneous detection of Listeria monocytogenes and Staphylococcus aureus. The primers were designed for the specific genes hlyA of L. monocytogenes and nuc of S. aureus, respectively, and the reaction conditions were optimized. The specificity and sensitivity of the method were also evaluated. Results showed that L. monocytogenes and S. aureus could be detected simultaneously in 60 minutes, and the detection limits were 7.3×102 and 2.3×102 copies/mL, respectively. The method was of good specificity and sensitivity, and the detection limits of 2.8×104 and 2.4×104 CFU/mL could be achieved in real samples. This dual DARQ-LAMP assay could detect two kinds of foodborne pathogens simultaneously, which would shorten the detection time and improve the detection efficiency, providing a new idea for the study of multiple LAMP detection of foodborne pathogens.

Key words： foodborne pathogens; Listeria monocytogenes; Staphylococcus aureus; detection of amplification by release of quenching and loop-mediated isothermal amplification (DARQ-LAMP); rapid detection

Cite this article

CHEN Qingying , LIU Dan , ZHANG Xingguo , WEI Jinyuan , ZHONG Qingping . Simultaneous detection of Listeria monocytogenes and Staphylococcus aureus based on dual DARQ-LAMP method[J]. Food and Fermentation Industries, 2023 , 49(22) : 279 -286 . DOI: 10.13995/j.cnki.11-1802/ts.035031

References

[1] 马佳睿, 谢婧, 李瑞乾, 等. LAMP在食源性致病菌检测中的应用进展[J]. 黑龙江畜牧兽医, 2021(17):39-43.
MA J R, XIE J, LI R Q, et al. Application progress on LAMP in detection of food-borne pathogens[J]. Heilongjiang Animal Science and Veterinary Medicine, 2021(17):39-43.
[2] ZHAO X H, LIN C W, WANG J, et al. Advances in rapid detection methods for foodborne pathogens[J]. Journal of Microbiology and Biotechnology, 2014, 24(3):297-312.
[3] LAW J W F, AB MUTALIB N S, CHAN K G, et al. Rapid methods for the detection of foodborne bacterial pathogens: Principles, applications, advantages and limitations[J]. Frontiers in Microbiology, 2015, 5:770.
[4] 刘洪蕾, 王真. 食源性单核细胞增生李斯特氏菌检测方法研究进展[J]. 动物医学进展, 2022, 43(10):111-116.
LIU H L, WANG Z. Progress on detection methods of foodborne Listeria monocytogenes[J]. Progress in Veterinary Medicine, 2022, 43(10):111-116.
[5] NOTOMI T, OKAYAMA H, MASUBUCHI H, et al. Loop-mediated isothermal amplification of DNA[J]. Nucleic Acids Research, 2000, 28(12): e63.
[6] LI Y M, FAN P H, ZHOU S S, et al. Loop-mediated isothermal amplification (LAMP): A novel rapid detection platform for pathogens[J]. Microbial Pathogenesis, 2017, 107:54-61.
[7] LEE J E, MUN H, KIM S R, et al. A colorimetric Loop-mediated isothermal amplification (LAMP) assay based on HRP-mimicking molecular beacon for the rapid detection of Vibrio parahaemolyticus[J]. Biosensors and Bioelectronics, 2020, 151:111968.
[8] ARUTSELVAN R, REDDY M K, MAKESHKUMAR T. Rapid detection of tomato leaf curl Bengaluru virus through loop mediated isothermal amplification assay[J]. VirusDisease, 2017, 28(3):303-308.
[9] YEASMIN S, TAKABATAKE R, KAGIYA Y, et al. Evaluation of a species-specific, stable, and endogenous sequence of eggplant (Solanum melongena) using LAMP for the detection of genetically modified eggplants[J]. Food Hygiene and Safety Science, 2021, 62(6):180-186.
[10] MAYBORODA O, KATAKIS I, O′SULLIVAN C K. Multiplexed isothermal nucleic acid amplification[J]. Analytical Biochemistry, 2018, 545:20-30.
[11] OSCORBIN I P, BELOUSOVA E A, ZAKABUNIN A I, et al. Comparison of fluorescent intercalating dyes for quantitative loop-mediated isothermal amplification (qLAMP)[J]. BioTechniques, 2016, 61(1):20-25.
[12] QUYEN T L, NGO T A, BANG D D, et al. Classification of multiple DNA dyes based on inhibition effects on real-time loop-mediated isothermal amplification (LAMP): Prospect for point of care setting[J]. Frontiers in Microbiology, 2019, 10:2234.
[13] CHUANG T L, WEI S C, LEE S Y, et al. A polycarbonate based surface plasmon resonance sensing cartridge for high sensitivity HBV loop-mediated isothermal amplification[J]. Biosensors and Bioelectronics, 2012, 32(1):89-95.
[14] ALMASI M A, ALMASI G. Colorimetric immunocapture loop mediated isothermal amplification assay for detection of Impatiens necrotic spot virus (INSV) by GineFinderTM dye[J]. European Journal of Plant Pathology, 2018, 150(2):533-538.
[15] TANNER N A, ZHANG Y H, EVANS T C Jr. Simultaneous multiple target detection in real-time loop-mediated isothermal amplification[J]. BioTechniques, 2012, 53(2):81-89.
[16] KÜHBACHER A, NOVY K, QUEREDA J J, et al. Listeriolysin O-dependent host surfaceome remodeling modulates Listeria monocytogenes invasion[J]. Pathogens and Disease, 2018, 76(8): fty082.
[17] HOEGH S V, SKOV M N, BOYE K, et al. Variations in the Staphylococcus aureus-specific nuc gene can potentially lead to misidentification of meticillin-susceptible and-resistant S. aureus[J]. Journal of Medical Microbiology, 2014, 63(7):1020-1022.
[18] 张小雨, 王雯雯, 马臣杰, 等. 金黄色葡萄球菌LAMP检测方法的建立及应用[J]. 农业生物技术学报, 2021, 29(9):1845-1854.
ZHANG X Y, WANG W W, MA C J, et al. Establishment and application of loop-mediated isothermal amplification assay for detection of Staphylococcus aureus[J]. Journal of Agricultural Biotechnology, 2021, 29(9):1845-1854.
[19] 刘丹, 谢加玲, 张孟雨, 等. 基于实时荧光环介导等温扩增技术同时检测副溶血弧菌与金黄色葡萄球菌[J]. 食品与发酵工业, 2022, 48(22):255-262.
LIU D, XIE J L, ZHANG M Y, et al. Simultaneous detection of Vibrio parahaemolyticus and Staphylococcus aureus by real-time LAMP[J]. Food and Fermentation Industries, 2022, 48(22):255-262.
[20] 纪懿芳, 胡文忠, 姜爱丽, 等. LAMP技术快速检测海产品副溶血弧菌的条件优化[J]. 食品工业科技, 2015, 36(20):59-63; 67.
JI Y F, HU W Z, JIANG A L, et al. Optimize of Loop-mediated isothermal amplification technology in detection of Vibrio parahaemolyticus[J]. Science and Technology of Food Industry, 2015, 36(20):59-63; 67.
[21] 胡元庆, 沈子晨, 李凤霞, 等. 基于blaCARB-17基因建立水产品中副溶血弧菌的环介导等温扩增技术检测方法[J]. 食品与发酵工业, 2020, 46(23):198-206.
HU Y Q, SHEN Z C, LI F X, et al. blaCARB-17-based LAMP assay for detecting Vibrio parahaemolyticus in aquatic products[J]. Food and Fermentation Industries, 2020, 46(23):198-206.
[22] ZHANG L, ZENG J, MA D, et al. Application and evaluation of loop-mediated isothermal amplification method for detecting of Listeria monocytogenes in food[J]. Chinese Journal of Preventive Medicine, 2014, 48(3): 213-217.
[23] NATHANIEL B R, GHAI M, DRUCE M, et al. Development of a loop-mediated isothermal amplification assay targeting lmo0753 gene for detection of Listeria monocytogenes in wastewater[J]. Letters in Applied Microbiology, 2019, 69(4):264-270.
[24] WANG C, LI R, QUAN S, et al. GMO detection in food and feed through screening by visual loop-mediated isothermal amplification assays[J]. Analytical and Bioanalytical Chemistry, 2015, 407(16):4829-4834.
[25] 徐文文, 梁玉林, 王云霞, 等. 二重环介导等温扩增法快速检测乳粉中沙门氏菌和金黄色葡萄球菌[J]. 食品与发酵工业, 2021, 47(2):241-246.
XU W W, LIANG Y L, WANG Y X, et al. Rapid detection of Salmonella and Staphylococcus aureus in milk powder by double loop-mediated isothermal amplification[J]. Food and Fermentation Industries, 2021, 47(2):241-246.
[26] DING X, YIN K, LI Z Y, et al. Cleavable hairpin beacon-enhanced fluorescence detection of nucleic acid isothermal amplification and smartphone-based readout[J]. Scientific Reports, 2020, 10:18819.
[27] NANAYAKKARA I A, WHITE I M. Demonstration of a quantitative triplex LAMP assay with an improved probe-based readout for the detection of MRSA[J]. The Analyst, 2019, 144(12):3878-3885.
[28] 冯可, 胡文忠, 姜爱丽, 等. 鲜切果蔬中4种病原微生物多重PCR检测技术[J]. 食品科学, 2018, 39(6):276-283.
FENG K, HU W Z, JIANG A L, et al. Multiplex PCR method for detection of four foodborne pathogens on fresh-cut fruits and vegetables[J]. Food Science, 2018, 39(6):276-283.
[29] DUARTE C, SALM E, DORVEL B, et al. On-chip parallel detection of foodborne pathogens using loop-mediated isothermal amplification[J]. Biomedical Microdevices, 2013, 15(5):821-830.
[30] 吕观, 常彦磊, 石磊. 免疫磁珠-环介导等温扩增快速检测牛肉中的鼠伤寒沙门氏菌与金黄色葡萄球菌[J]. 肉类研究, 2019, 33(7):42-48.
LÜ G, CHANG Y L, SHI L. Rapid detection of Salmonella typhimurium and Staphylococcus aureus in beef by immunomagnetic separation combined with loop-mediated IsothermaL amplification method[J]. Meat Research, 2019, 33(7):42-48.
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