Skip to main content

Advertisement

SpringerLink
Log in

Development and application of a loop-mediated isothermal amplification method on rapid detection Escherichia coli O157 strains from food samples

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

We developed and evaluated the specificity and sensitivity of a loop-mediated isothermal amplification (LAMP) method for rapid detection of the food-borne Escherichia coli O157 strains. Six primers, including outer primers, inner primers and loop primers, were specially designed for recognizing eight distinct sequences on three targets, which were rfbE, stx1 and stx2. The detection limits were found to be 100, 100 and 10 fg DNA/tube for rfbE, stx1 and stx2, respectively. Application of LAMP assays were performed on 417 food-borne E. coli strains, the sensitivity of LAMP assays for the rfbE, stx1 and stx2 was 100, 95.3 and 96.3%, and the negative predictive value was 100, 96.7 and 97.1%, respectively; with a 100% specificity and positive predictive value for all three targets.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Baudouin V, Mariani-Kurkdjian P, Macher MA, Ait-Ifrane S, Garnier A, Bingen E, Loirat C, Deschenes G (2008) Treatment of a relapsing HUS secondary to prolonged carriage of E. coli O157: H7. Pediatr Nephrol 23:1599

    Google Scholar 

  2. Chapman PA, Ellin M, Ashton R (2001) A comparison of immunomagnetic separation and culture, Reveal(TM) and VIPTM for the detection of E. coli O157 in enrichment cultures of naturally-contaminated raw beef, lamb and mixed meat products. Lett Appl Microbiol 32:171–175

    Article  CAS  PubMed  Google Scholar 

  3. Crawford CG, Wijey C, Fratamico P, Tu SI, Brewster J (2000) Immunomagnetic-electrochemiluminescent detection of E. coli O157: H7 in ground beef. J Rapid Methods Autom Microbiol 8:249–264

    Article  CAS  Google Scholar 

  4. Gehring AG, Tu SI (2005) Enzyme-linked immunomagnetic electrochemical detection of live Escherichia coli O157: H7 in apple juice. J Food Protect 68:146–149

    CAS  Google Scholar 

  5. Horisaka T, Fujita K, Iwata T, Nakadai A, Okatani AT, Horikita T, Taniguchi T, Honda E, Yokomizo Y, Hayashidani H (2004) Sensitive and specific detection of Yersinia pseudotuberculosis by loop-mediated isothermal amplification. J Clin Microbiol 42:5349–5352

    Article  CAS  PubMed  Google Scholar 

  6. Jung BY, Jung SC, Kweon CH (2005) Development of a rapid immunochromatographic strip for detection of Escherichia coli O157. J Food Protect 68:2140–2143

    CAS  Google Scholar 

  7. Kaneko H, Kawana T, Fukushima E, Suzutani T (2007) Tolerance of loop-mediated isothermal amplification to a culture medium and biological substances. J Biochem Biophys Methods 70:499–501

    Article  CAS  PubMed  Google Scholar 

  8. Mori Y, Nagamine K, Tomita N, Notomi T (2001) Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun 289:150–154

    Article  CAS  PubMed  Google Scholar 

  9. Mullah B, Livak K, Andrus A, Kenney P (1998) Efficient synthesis of double dye-labeled oligodeoxyribonucleotide probes and their application in a real time PCR assay. Nucleic Acids Res 26:1026–1031

    Article  CAS  PubMed  Google Scholar 

  10. Nagamine K, Watanabe K, Ohtsuka K, Hase T, Notomi T (2001) Loop-mediated isothermal amplification reaction using a nondenatured template. Clin Chem 47:1742–1743

    CAS  PubMed  Google Scholar 

  11. Organization, World Health (2007) Food safety and foodborne illness. Available via DIALOG. http://www.who.int/mediacentre/factsheets/fs237/en/print.html

  12. Parida M, Horioke K, Ishida H, Dash PK, Saxena P, Jana AM, Islam MA, Inoue S, Hosaka N, Morita K (2005) Rapid detection and differentiation of dengue virus serotypes by a real-time reverse transcription-loop-mediated isothermal amplification assay. J Clin Microbiol 43:2895–2903

    Article  CAS  PubMed  Google Scholar 

  13. Rasekh J, Thaler AM, Engeljohn DL, Pihkala NH (2005) Food safety and inspection service policy for control of poultry contaminated by digestive tract contents: a review. J Appl Poult Res 14:603–611

    Google Scholar 

  14. Sanchez A, Navarro RB, Marquez RF, Quintana MF, Perez LS, Guilarte OD, Martinez HB, Bello LM (2008) Enterohemorrhagic Escherichia coli O157: H7 isolates from children in Cuba. Pediatr Infect Dis J 27:1122–1123

    Article  PubMed  Google Scholar 

  15. Shelton DR, Karns JS (2001) Quantitative detection of Escherichia coli O157 in surface waters by using immunomagnetic electrochemiluminescence. Appl Environ Microbiol 67:2908–2915

    Article  CAS  PubMed  Google Scholar 

  16. Su JY, Shi L, Yang LS, Xiao ZH, Li XH, Yamasaki S (2006) Analysis of integrons in clinical isolates of Escherichia coli in China during the last six years. FEMS Microbiol Lett 254:75–80

    Article  CAS  PubMed  Google Scholar 

  17. Tsugunori Notomi HOHM (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28:E63

    Article  Google Scholar 

  18. Tu SI, Golden M, Fett WF, Gehring A, Irwin P (2003) Rapid detection of outbreak Escherichia coli O157 and Salmonella on alfalfa sprouts by immunomagnetic capture and time-resolved fluorescence. J Food Saf 23:75–89

    Article  Google Scholar 

  19. Wang L, Li L, Alam MJ, Geng YH, Li ZY, Yamasaki SJ, Shi L (2008) Loop-mediated isothermal amplification method for rapid detection of the toxic dinoflagellate Alexandrium, which causes algal blooms and poisoning of shellfish. FEMS Microbiol Lett 282:15–21

    Article  CAS  PubMed  Google Scholar 

  20. Wang L, Shi L, Alam MJ, Geng Y, Li L (2008) Specific and rapid detection of foodborne Salmonella by loop-mediated isothermal amplification method. Food Res Int 41:69–74

    Article  Google Scholar 

  21. Xu Z, Shi L, Zhang C, Zhang L, Li X, Cao Y, Li L, Yamasaki S (2007) Nosocomial infection caused by class 1 integron-carrying Staphylococcus aureus in a hospital in South China. Clin Microbiol Infect 13:980–984

    Article  CAS  PubMed  Google Scholar 

  22. Xu ZB, Li L, Alam MJ, Zhang LY, Yamasaki S, Shi L (2008) First confirmation of integron-bearing methicillin-resistant Staphylococcus aureus. Current Microbiology 57:264–268

    Article  CAS  PubMed  Google Scholar 

  23. Xu ZB, Li L, Shirtliff ME, Alam MJ, Yamasaki S, Shi L (2009) Occurrence and characteristics of class 1 and 2 integrons in Pseudomonas aeruginosa isolates from patients in Southern China. J Clin Microbiol 47:230–234

    Article  CAS  PubMed  Google Scholar 

  24. Xu ZB, Shi L, Alam MJ, Li L, Yamasaki S (2008) Integron-bearing methicillin-resistant coagulase-negative staphylococci in South China, 2001–2004. FEMS Microbiol Lett 278:223–230

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by Science Foundation of Ministry of Education of China(706046), National Natural Science Foundation of China (20436020) and State Scholarship Fund of China Scholarship Council (2008615044).

Author information

Authors and Affiliations

  1. College of Light Industry and Food Sciences, South China University of Technology, 510640, Guangzhou, China

    Xihong Zhao, Lijun You, Zhenbo Xu, Lin Li, Xiaowei He & Liansheng Yang

  2. Guangzhou Medical College, Guangzhou, China

    Yanmei Li

  3. Food Safety Key Laboratory of Guangdong Province, College of Food Science, South China Agricultural University, Guangzhou, China

    Li Wang

  4. Department of Microbial Pathogenesis, Dental School, University of Maryland-Baltimore, Baltimore, MD, 21201, USA

    Zhenbo Xu

  5. Zhongshan Supervision Testing Institute of Quality & Metrology, Zhongshan, China

    Yao Liu

  6. Guangzhou Wondfo Biotechnology Company, Guangzhou, China

    Jihua Wang

Authors
  1. Xihong Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yanmei Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Li Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lijun You
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhenbo Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Lin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Xiaowei He
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yao Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jihua Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Liansheng Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Zhenbo Xu or Xiaowei He.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhao, X., Li, Y., Wang, L. et al. Development and application of a loop-mediated isothermal amplification method on rapid detection Escherichia coli O157 strains from food samples. Mol Biol Rep 37, 2183–2188 (2010). https://doi.org/10.1007/s11033-009-9700-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-009-9700-6

Keywords

Search

Navigation