Agricultural Research

, Volume 8, Issue 4, pp 490–496 | Cite as

Rapid Detection of Shiga toxin-Producing E. Coli in Animal Origin Foods Using Loop-Mediated Isothermal Amplification (LAMP) Assay

  • P. Baraily
  • R. J. Zende
  • D. P. KshirsagarEmail author
  • V. M. Vaidya
  • R. N. Waghamare
  • A. M. Paturkar
  • R. P. Todankar
  • A. H. Shirke
Full-Length Research Article


The aim of this study was comparative evaluation of loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) assay for rapid and inexpensive detection of shiga toxin-producing E. coli in animal origin foods by targeting stx1 and stx2 genes. The LAMP assay was performed using a water bath. The standardized LAMP assay was evaluated on 122 E. coli field isolates obtained from various animal origin food samples to ensure its reliability and usefulness. The result showed that conventional PCR could detect 68 (55.73%) and 75 (61.47%) positive E. coli isolates for stx1 and stx2 genes. Whereas, LAMP showed higher sensitivity by detecting 79 (64.75%) and 87 (71.31%) positive isolates of E. coli for stx1 and stx2 genes, respectively. LAMP assay was found to be highly specific and 10 times more sensitive as it could detect 1.11 × 102 cfu/ml for both stx1 and stx2 genes of E. coli isolates, whereas conventional PCR could detect 1.85 x 103 cfu/ml for both stx1 and stx2 genes of E. coli isolates. The rapidness, sensitivity, specificity, easiness and cost-effectiveness of LAMP assays will be very useful for the detection of foodborne pathogens for improving food sanitation and maintaining food safety.


Loop-mediated isothermal amplification (LAMP) Shiga toxin-producing E. coli Food safety Polymerase chain reaction (PCR) Sensitivity Specificity 



This work was supported by grants from Indian Council of Agricultural Research, New Delhi, under the Project “All India Co-Ordinated Research Project on Post Harvest Engineering and Technology” implemented at Bombay Veterinary College, Parel, Mumbai.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Barman NN, Deb R, Ramamurthy T, Sharma RK, Borah P, Wani SA, Kalita D (2007) Molecular characterization of Shiga toxin producing E. coli isolates from pigs oedema. Indian J Med Res 127:602–606Google Scholar
  2. 2.
    Bettelheim KA (2000) Role of non-O157 VTEC. J Appl Microbiol 88:38–50CrossRefGoogle Scholar
  3. 3.
    Burns AJ, Herbert TM, Ward SM, Sanders KM (1997) Interstitial cells of Cajal in the guinea-pig gastrointestinal tract as revealed by C-Kit immunohistochemistry. Cell Tissue Res 290:11–20CrossRefGoogle Scholar
  4. 4.
    Hara-Kudo Y, Nemoto J, Ohtsuka K, Segawa Y, Takatori K, Kojima T, Ikedo M (2007) Sensitive and rapid detection of verotoxin producing E. coli using loop-mediated isothermal amplification, World health organization (WHO) Fact Sheet. J Med Microbiol 56:398–406CrossRefGoogle Scholar
  5. 5.
    Hazarika RA, Singh DK, Kapoor KN, Agrawal RK, Pandey AB, Purosottam (2007) Verotoxigenic E.coli (STEC) from beef and its products. Indian J Expt Biol 45:207–211Google Scholar
  6. 6.
    Hill J, Beriwal S, Chandra I, Paul VK, Kapil A, Singh AT, Wadowsky RM, Singh V, Goyal A, Jahnukainen T, Johnson JR, Tarr PI, Vats A (2008) Loop-mediated isothermal amplification assay for rapid detection of common strains of E. coli. J Clin Microbiol 46:2800–2804CrossRefGoogle Scholar
  7. 7.
    Mahony J, Chong S, Stone C, Chui L (2016) Evaluation of four loop-mediated isothermal amplification (LAMP) assays for identification of Shiga toxin producing E.coli O157 (STEC) and non-O157 Strains. Adv Mol Diag 1(1):3–6Google Scholar
  8. 8.
    Matise I, Shelton M, Phillips G and Will LA (1998). Sensitive PCR method for detection of E.coli 0157:H7 and other Shiga toxin-producing bacteria in ground meat. Department of Microbiol, Immunol and Preventive Medicine, Iowa State University. paper- 33Google Scholar
  9. 9.
    Nataro JP, Kaper JB (1998) Diarrheagenic E. coli. Clin Microbiol Rev 11:142–201CrossRefGoogle Scholar
  10. 10.
    Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28:63CrossRefGoogle Scholar
  11. 11.
    Ohtsuka K, Yanagawa K, Takatori K, Hara-Kudo Y (2005) Detection of Salmonella enterica in naturally contaminated liquid eggs by Loop-mediated Isothermal Amplification, and characterization of Salmonella isolates. Appl Environ Microbiol 71(11):6730–6735CrossRefGoogle Scholar
  12. 12.
    Rawool DB, Malik SVS, Barbuddhe SB, Shakuntala I, Aurora R (2007) A multiplex PCR for detection of virulence associated genes in Listeria monocytogenes. Int J Food Safety 9:56–62Google Scholar
  13. 13.
    Song T, Toma C, Nakasone N, Iwanaga M (2005) Sensitive and rapid detection of Shigella and enteroinvasive E.coli by a loop-mediated isothermal amplification method. FEMS Microbiol. 243:259–263CrossRefGoogle Scholar
  14. 14.
    Stratakos AC, Linton M, Millington S, Grant IR (2016) A loop-mediated isothermal amplification method for rapid direct detection and differentiation of non-pathogenic and verocytotoxigenic E. coli in beef and bovine faeces. J Appl Microbiol 122:817–828CrossRefGoogle Scholar
  15. 15.
    Tang T, Cheng A, Wang M, Li X, He Q, Jia R, Zhu D, Chen X (2012) Development and clinical verification of a loop-mediated isothermal amplification method for detection of Salmonella species in suspect infected ducks. Poult Sci 91:979–986CrossRefGoogle Scholar
  16. 16.
    Teh CSJ, Chua KH, Lim YAL, Lee SC, Thong KL (2014) Loop-mediated isothermal amplification assay for detection of generic and vero cytotoxin producing E. coli among indigenous individuals in Malaysia. Hindawi Publishing Corporation Scientific World Journal 2014:6. CrossRefGoogle Scholar
  17. 17.
    Tenaillon O, Skurnik D, Picard B, Denamur E (2010) The population genetics of commensal Escherichia coli. Nat Rev Microbiol 8:207–217CrossRefGoogle Scholar
  18. 18.
    Verma S, Kumar M, Kashyap S, Singh M, Venkatesh V (2013) Current scenario of E. coli and its serotype “O157:H7” in Indian subcontinent. Int J Innovative Res Sci Eng Technol 2(7):2642–2644Google Scholar
  19. 19.
    Vidal M, Kruger E, Duran C, Lagos R, Levine M, Prado V, Toro C, Vidal R (2005) Single multiplex PCR assay to identify simultaneously the six categories of diarrheagenic E.coli associated with enteric Infections. J Clin Microbiol 24:5362–5365CrossRefGoogle Scholar
  20. 20.
    Wang D, Liu F, Huo G, Ren D, Li Y (2009) Development and evaluation of loop- mediated isothermal amplification method for detecting E. coli O157 in raw milk. J Rapid Methods Autom Microbiol 17:55–66CrossRefGoogle Scholar
  21. 21.
    Wang F, Jiang L, Ge B (2011) Loop-mediated isothermal amplification assays for detecting shiga toxin-producing E. coli in ground beef and human stools. J Clin Microbiol 50:91–97CrossRefGoogle Scholar
  22. 22.
    Wang F, Jiang L, Yang Q, Prinyawiwatkul W, Ge B (2012) Rapid and specific detection of E. coli serogroups O26, O45, O103, O111, O121, O145 and O157 in ground beef, beef trim produce by loop-mediated isothermal amplification. Appl Environ Microbiol 78(8):2727–2736. CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    World Health Organization (2007) Food safety–foodborne diseases and value chain management for food safety. (“Forging links between agriculture and health” CGIAR on agriculture and health meeting in WHO/HQ)Google Scholar
  24. 24.
    Zende R, Kshirsagar D, Vaidya V, Waghamare R, Todankar R, Shirke A (2017) Loop-mediated isothermal amplification assay (LAMP): a rapid tool for diagnosis of foodborne and zoonotic pathogens: a review. Int J Livest Res 7(5):23–35Google Scholar

Copyright information

© NAAS (National Academy of Agricultural Sciences) 2018

Authors and Affiliations

  • P. Baraily
    • 1
    • 2
  • R. J. Zende
    • 1
    • 2
  • D. P. Kshirsagar
    • 1
    • 2
    Email author
  • V. M. Vaidya
    • 1
    • 2
  • R. N. Waghamare
    • 1
    • 2
  • A. M. Paturkar
    • 1
    • 2
  • R. P. Todankar
    • 1
    • 2
  • A. H. Shirke
    • 1
    • 2
  1. 1.Department of Veterinary Public Health & EpidemiologyBombay Veterinary CollegeParel, MumbaiIndia
  2. 2.Maharashtra Animal and Fishery Science UniversityNagpurIndia

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