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Loop-mediated isothermal amplification (LAMP): a sensitive molecular tool for detection of Staphylococcus aureus in meat and dairy product

  • Food Microbiology - Research Paper
  • Published:
Brazilian Journal of Microbiology Aims and scope Submit manuscript

Abstract

Staphylococcus aureus–mediated food poisoning is a primary concern worldwide. The presence of the organism in food is an indicative of poor sanitation during production, and it is essential to have efficient methods for detecting this pathogen. A novel molecular diagnostic technique called loop-mediated isothermal amplification (LAMP) serves as a rapid and sensitive detection method, which amplifies nucleic acids at isothermal conditions. In this study, a LAMP-based diagnostic assay was developed to detect Staphylococcus aureus (S. aureus) using two target genes femA and arcC. The optimum reaction temperature was found to be 65 °C and at 60 °C for femA and arcC genes, respectively. The developed assay specifically amplified DNA from S. aureus, not from other related bacterial species and compared to PCR, and a 100-fold higher sensitivity was observed. Furthermore, the LAMP assay could detect the pathogen from food samples mainly meat and dairy samples when analyzed in both intact and enriched conditions. Thirteen samples were found positive for S. aureus with LAMP showing a greater number of positive samples in comparison to PCR. This study established a highly sensitive and a rapid diagnostic procedure for the detection and surveillance of this major foodborne pathogen.

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References

  1. Le Loir Y, Baron F, Gautier M (2003) Staphylococcus aureus and food poisoning. Genet Mol Res 2(1):63–76

    PubMed  Google Scholar 

  2. Mead PS, Slutsker L, Dietz V, McCaig LF, Bresee JS, Shapiro C, Griffin PM, Tauxe RV (1999) Food-related illness and death in the United States.". Emerg Infect Dis 5(5):607

    Article  CAS  Google Scholar 

  3. De Boer E, Zwartkruis-Nahuis JT, Wit B, Huijsdens XW, De Neeling AJ, Bosch T, Van Oosterom RA, Vila A, Heuvelink AE (2011) Prevalence of methicillin-resistant Staphylococcus aureus in meat. Int J Food Microbiol 134:52–56

    Article  Google Scholar 

  4. Ausubel FM, Kingston R, Moore RE, Seidman DD, Smith, J G JA, Struhl K, Ferreira Behravesh CB, Williams, IT, Tauxe RV (2012) Emerging foodborne pathogens and problems: expanding prevention efforts before slaughter or harvest. Improving Food Safety through a One Health Approach Workshop Summary.

  5. Dwivedi HP, Jaykus LA (2011) Detection of pathogens in foods: the current state-of-the-art and future directions. Crit Rev Microbiol 37:40–63

    Article  CAS  Google Scholar 

  6. Alarcon B, Vicedo B, Aznar R (2006) PCR-based procedures for detection and quantification of Staphylococcus aureus and their application in food. J Appl Microbiol 2:352–364

    Article  Google Scholar 

  7. Su J, Liu X, Cu H, Li Y, Chen D, Li Y, Yu G (2014) Rapid and simple detection of methicillin-resistance staphylococcus aureus by orfX loop-mediated isothermal amplification assay. BMC Biotechnol 14:8

    Article  Google Scholar 

  8. Rodríguez-Lázaro D, Oniciuc EA, García PG, Gallego D, Fernández-Natal I, Dominguez-Gil M, Eiros-Bouza JM, Wagner M, Nicolau AI, Hernández M (2017) Detection and characterization of Staphylococcus aureus and methicillin-resistant S aureus in foods confiscated in EU borders. Frontiers in microbiology 8:1344

    Article  Google Scholar 

  9. Pereira V, Lopes C, Castro A, Silva J, Gibbs P, Teixeira P (2009) Characterization for enterotoxin production, virulence factors, and antibiotic susceptibility of Staphylococcus aureus isolates from various foods in Portugal. Food Microbiol 26(3):278–282

    Article  CAS  Google Scholar 

  10. Goto M, Hayashidani H, Takatori K, Hara-Kudo Y (2007) Rapid detection of enterotoxigenic Staphylococcus aureus harbouring genes for four classical enterotoxins, SEA, SEB, SEC and SED, by loop-mediated isothermal amplification assay. Lett Appl Microbiol 45:100–107

    Article  CAS  Google Scholar 

  11. Byrne B, Gilmartin N, Lakshmanan RS, O’Kennedy R (2015) Antibodies, enzymes, and nucleic acid sensors for high throughput screening of microbes and toxins in food. High throughput screening for food safety assessment: 25–80.

  12. Hennekinne JA, De Buyser ML, Dragacci S (2012) Staphylococcus aureus and its food poisoning toxins: characterization and outbreak investigation. FEMS microbiology reviews 36:815–836

    Article  CAS  Google Scholar 

  13. Tomita N, Mori Y, Kanda H, Notomi T (2008) Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products. Nat Protoc 3:877–882

    Article  CAS  Google Scholar 

  14. Sowmya N, Thakur MS, Manonmani HK (2012) Rapid and simple DNA extraction method for the detection of enterotoxigenic Staphylococcus aureus directly from food samples: comparison of PCR and LAMP methods. Journal of applied microbiology 113:106–113

    Article  CAS  Google Scholar 

  15. Shriver-Lake LC, Shubin YS, Ligler FS (2003) Detection of staphylococcal enterotoxin B in spiked food samples. J Food Prot 66:1851–1856

    Article  CAS  Google Scholar 

  16. Sudershan RV, Naveen Kumar R, Kashinath L, Bhaskar V, Polasa K (2014) Foodborne infections and intoxications in Hyderabad India. Epidemiology Research International, 2014.

  17. Straub TM, Chandler DP (2003) Towards a unified system for detecting waterborne pathogens. J Microbiol Methods 53:185–197

    Article  Google Scholar 

  18. Koreen L, Ramaswamy SV, Graviss EA, Naidich S, Musser JM, Kreiswirth BN (2004) spa typing method for discriminating among Staphylococcus aureus isolates: implications for use of a single marker to detect genetic micro-and macro variation. J Clin Microbiol 42:792–799

    Article  CAS  Google Scholar 

  19. Girish PS, Anjaneyulu ASR, Viswas KN, Shivakumar BM, Anand M, Patel M, Sharma B (2005) Meat species identification by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of mitochondrial 12S rRNA gene. Meat Sci 70:107–112

    Article  CAS  Google Scholar 

  20. Johnson WM, Tyler SD, Ewan EP, Ashton FE, Pollard DR, Rozee KR (1991) Detection of genes for enterotoxins, exfoliative toxins, and toxic shock syndrome toxin 1 in Staphylococcus aureus by the polymerase chain reaction. J Clin Microbiol 29(3):426–430

    Article  CAS  Google Scholar 

  21. Omoe K, Ishikawa M, Shimoda Y, Hu DL, Ueda S, Shinagawa K (2002) Detection of seg, seh, and sei genes in Staphylococcus aureus isolates and determination of the enterotoxin productivities of S. aureus isolates harbouring seg, seh, or sei genes. J Clin Microbiol 40:857–862

    Article  CAS  Google Scholar 

  22. Pereira V, Lopes C, Castro A, Silva J, Gibbs P, Teixeira P (2009) Characterization for enterotoxin production, virulence factors, and antibiotic susceptibility of Staphylococcus aureus isolates from various foods in Portugal. Food Microbiol 26:278–282

    Article  CAS  Google Scholar 

  23. Zhang T, Wang C, Wei X, Zhao X, Zhong X (2012) Loop-mediated isothermal amplification for detection of Staphylococcus aureus in dairy cow suffering from mastitis. Journal of Biomedicine and Biotechnology: 435982.

  24. Su J, Liu X, Cui H, Li Y, Chen D, Li Y, Yu G (2014) Rapid and simple detection of methicillin-resistance staphylococcus aureus by orfX loop-mediated isothermal amplification assay. BMC Biotechnol 14:8

    Article  Google Scholar 

  25. Koide Y, Maeda H, Yamabe K, Naruishi K, Yamamoto T, Kokeguchi S, Takashiba S (2010) Rapid detection of mecA and spa by the loop-mediated isothermal amplification (LAMP) method. Lett Appl Microbiol 50(4):386–392. https://doi.org/10.1111/j.1472-765X.2010.02806.x

    Article  CAS  PubMed  Google Scholar 

  26. Chen C, Zhao Q, Guo J, Li Y, Chen Q (2017) Identification of methicillin-resistant Staphylococcus aureus (MRSA) using simultaneous detection of mecA. nuc. and femB by loop-mediated isothermal amplification (LAMP). Curr Microbiol 74(8):965–971. https://doi.org/10.1007/s00284-017-1274-2

    Article  CAS  PubMed  Google Scholar 

  27. Sowmya N, Thakur M, Manonmani H (2012) Rapid and simple DNA extraction method for the detection of enterotoxigenic Staphylococcus aureus directly from food samples: comparison of PCR and LAMP methods. J Appl Microbiol 113:106–113. https://doi.org/10.1111/j.1365-2672.2012.05315.x

    Article  CAS  PubMed  Google Scholar 

  28. Hu DL, Li S, Fang R, Ono HK (2021) Update on molecular diversity and multipathogenicity of staphylococcal superantigen toxins. Animal Diseases 1:7. https://doi.org/10.1186/s44149-021-00007-7

    Article  Google Scholar 

  29. Li Y, Yang L, Fu J, Yan M, Chen D, Zhang L (2017) The novel loop-mediated isothermal amplification based confirmation methodology on the bacteria in Viable but Non-Culturable (VBNC) state. Microb Pathog 111:280–284. https://doi.org/10.1016/j.micpath.2017.09.007

    Article  CAS  PubMed  Google Scholar 

  30. Pang B, Yao S, Xu K, Wang J, Song X, Mu Y, Zhao C, Li J (2019) A novel visual-mixed-dye for LAMP and its application in the detection of foodborne pathogens. Anal Biochem 574:1–6. https://doi.org/10.1016/j.ab.2019.03.002

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We are grateful to the Department of Science and Technology (DST), Government of India for providing the fund under women Scientist Scheme (WOS-A): SR/WOS-A/LS-1065/2014 (G). The corresponding author also sincerely acknowledged Prof. Dr. Indrani Karunasagar for her constant guidance throughout the project.

Funding

Women Scientist Scheme (WOS-A): SR/WOS-A/LS-1065/2014 (G). Department of Science and Technology (DST), Government of India.

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Authors and Affiliations

  1. Nitte University Centre for Science Education and Research, Nitte (Deemed To Be University), Mangalore, India

    Manal Hassan, Rajeshwari Vittal, Juliet Mohan Raj & Gunimala Chakraborty

  2. Division of Molecular Genetics and Cancer, Nitte University Centre for Science Education and Research Paneer Campus, Nitte (Deemed To Be University), Karnataka, 575 018, Mangalore, India

    Gunimala Chakraborty

Authors
  1. Manal Hassan
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  2. Rajeshwari Vittal
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  3. Juliet Mohan Raj
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  4. Gunimala Chakraborty
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Contributions

MH: conducts the research and investigation process, specifically performing the experiments, or data collection, and manuscript preparation.

RV: conducts the research and investigation process.

JMR: experiments design and conducts the research and investigation process and manuscript preparation.

GC: conceptualization of the research design, funding acquisition, and manuscript preparation and supervision of the research.

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Correspondence to Gunimala Chakraborty.

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The authors declare no competing interests.

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Hassan, M., Vittal, R., Raj, J.M. et al. Loop-mediated isothermal amplification (LAMP): a sensitive molecular tool for detection of Staphylococcus aureus in meat and dairy product. Braz J Microbiol 53, 341–347 (2022). https://doi.org/10.1007/s42770-021-00659-0

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  • DOI: https://doi.org/10.1007/s42770-021-00659-0

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