European Food Research and Technology

, Volume 232, Issue 5, pp 769–776

Development and evaluation of a loop-mediated isothermal amplification assay for the rapid detection of Staphylococcus aureus in food

  • Hong Yang
  • Xiaoyan Ma
  • Xianzhou Zhang
  • Yu Wang
  • Wei Zhang
Original Paper

Abstract

A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Staphylococcus aureus in food was developed and evaluated. Heat-stable nuclease (nuc) gene was the target sequence amplified by LAMP. LAMP includes primer design, amplification at 64 °C, macroscopic observation, and estimation of the results. The LAMP detection of S. aureus was sensitive up to 1.25 CFU per reaction tube, with a detection limit of 10.3 CFU per reaction tube in the artificial contamination test, and the experiment took less than 1 h. Sensitivity of the method in Staphylococcus aureus detection was 100%, with 97.93% specificity and a 98.5% match ratio when compared using Chinese National Standard. The LAMP method is superior to the Chinese National Standard method in terms of sensitivity, cost, testing time, and convenience.

Keywords

Loop-mediated isothermal amplification LAMP Detection Staphylococcus aureus Food 

References

  1. 1.
    Tirado C, Schimdt K (2001) WHO surveillance programme for control of food-borne infections and intoxications: preliminary results and trends across greater Europe. J Infect 43:80–84Google Scholar
  2. 2.
    Atanassova V, Meindl A, Ring C (2001) Prevalence of Staphylococcus aureus and staphylococcal enterotoxins in raw pork and uncooked smoked ham-a comparison of classical culturing detection and RFLP-PCR. Int J Food Microb 68:105–113CrossRefGoogle Scholar
  3. 3.
    Chen Y (2007) A loop-mediated isothermal amplification method for detection of Staphylococcus aureus. Tianjin University, ChinaGoogle Scholar
  4. 4.
    Balaban N, Rasooly A (2000) Staphylococcal enterotoxins. Int J Food Microbiol 61(1):1–10CrossRefGoogle Scholar
  5. 5.
    Le Loir Y, Baron F, Gautier M (2003) Staphylococcus aureus and food poisoning. Genet Mol Res 2(1):63–76Google Scholar
  6. 6.
    Martin MC, Fueyo JM, Gonzalez-Hevia MA et al (2004) Genetic procedures for identification of enterotoxigenic strains of Staphylococcus aureus from three food poisoning outbreaks. Int J Food Microbiol 94(3):279–286CrossRefGoogle Scholar
  7. 7.
    Shriver-Lake LC, Shunbin YS, Ligler FS (2003) Detection of Staphylococcal enterotoxin B in spiked food samples. J Food Prot 66(10):1851–1856Google Scholar
  8. 8.
    Notomi T, Okayama H, Masubuchi H et al (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28(12):E63–E63CrossRefGoogle Scholar
  9. 9.
    Nagamine K, Watanabe K, Ohtsuka K et al (2001) Loop-mediated isothermal amplification reaction using a nondenatured template. Clin Chem 47(9):1742–1743Google Scholar
  10. 10.
    Mori Y, Nagamine K, Tomita N et al (2001) Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun 289(1):150–154CrossRefGoogle Scholar
  11. 11.
    Eiken Chemical Co. Ltd (2003) The principles of LAMP method. http://loopamp.eiken.co.ip/e/tech/index.html
  12. 12.
    Mori Y, Hirano T, Notomi T (2006) Sequence specific visual detection of LAMP reactions by addition of cationic polymers. BMC Biotechnol 6:3CrossRefGoogle Scholar
  13. 13.
    Nagamine K, Hase T, Notomi T (2002) Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol Cell Probes 16(3):223–229CrossRefGoogle Scholar
  14. 14.
    Ushio M, Yui I, Yoshida N et al (2005) Detection of respiratory syncytial virus genome by subgroups A, B specific reverse transcription loop-mediated isothermal amplification (RT-LAMP). J Med Virol 77(1):121–127CrossRefGoogle Scholar
  15. 15.
    Mori Y, Kitao M, Tomita N et al (2004) Real-time turbidimetry of LAMP reaction for quantifying template DNA. J Biochem Biophys Methods 59(2):145–157CrossRefGoogle Scholar
  16. 16.
    Okafuji T, Yoshida N, Fujino M et al (2005) Rapid diagnostic method for detection of mumps virus genome by loop-mediated isothermal amplification. J Clin Microbiol 43(4):1625–1631CrossRefGoogle Scholar
  17. 17.
    Fukuda S, Takao S, Kuwayama M et al (2006) Rapid detection of norovirus from specimens by real-time reverse transcription-loop-mediated isothermal amplification assay. J Clin Microbiol 44(4):1376–1381CrossRefGoogle Scholar
  18. 18.
    Dukes JP, King DP, Alex Andersen S (2006) Novel reverse transcription loop-mediated isothermal amplification for rapid detection of foot-and-mouth disease virus. Arch Virol 151(6):1093–1096CrossRefGoogle Scholar
  19. 19.
    Suzuki R, Yoshikawa T, Ihira M et al (2006) Development of the loop-mediated isothermal amplification method for rapid detection of cytomegalovirus DNA. J Virol Methods 132:216–221CrossRefGoogle Scholar
  20. 20.
    Song TY, Toma C, Nakasone N et al (2005) Sensitive and rapid detection of Shigella and enteroinvasive Escherichia coli by a loop-mediated isothermal amplification method. FEMS Microbiol Lett 243(2):259–263CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Hong Yang
    • 1
  • Xiaoyan Ma
    • 1
  • Xianzhou Zhang
    • 1
  • Yu Wang
    • 1
  • Wei Zhang
    • 1
  1. 1.College of Food Science and TechnologyAgricultural University of HeBeiBaodingPeople’s Republic of China

Personalised recommendations