Development of a filtration-based LAMP-LFA method as sensitive and rapid detection of O157:H7.

Jin-Hee Kim, Se-Wook Oh
Author Information
  1. Jin-Hee Kim: Department of Foods and Nutrition, Kookmin University, Seoul, 136-702 Korea.
  2. Se-Wook Oh: Department of Foods and Nutrition, Kookmin University, Seoul, 136-702 Korea. ORCID

Abstract

Rapid detection of low number of pathogenic bacteria in food is difficult. This study tested the filter-based loop-mediated isothermal amplification-lateral flow immunoassay (LAMP-LFA) method for rapid detection of pathogens in real food. O157:H7 was inoculated on 25 g of beef and the homogenized sample was filtered with 0.45 μm cellulose nitrate filter, and concentrated was recovered and DNA was extracted and analyzed by LAMP. LFA reaction was performed by hybridization of digoxygenin-labeled LAMP amplicon and biotinylated probe. The sensitivity of the filtered sample was 100 times more sensitive than that of the unfiltered sample. The total reaction time used for detection from sample preparation to confirmation of was within 3 h. These results suggest that the LAMP-LFA method can be used in real food systems as point-of-care testing for O157:H7 in beef.

Keywords

References

  1. J Food Prot. 1994 Apr;57(4):331-333 [PMID: 31113134]
  2. Mol Biol Rep. 2010 Jun;37(5):2183-8 [PMID: 19685165]
  3. J Food Sci. 2011 Jan-Feb;76(1):M41-6 [PMID: 21535692]
  4. Appl Environ Microbiol. 2002 Oct;68(10):4853-62 [PMID: 12324331]
  5. Int J Food Microbiol. 1992 Sep;17(1):37-45 [PMID: 1476866]
  6. J Food Sci. 2010 Apr;75(3):M165-72 [PMID: 20492306]
  7. Int J Food Microbiol. 2011 Jan 31;145(1):287-92 [PMID: 21276633]
  8. J Vet Sci. 2014;15(2):317-25 [PMID: 24675834]
  9. Appl Environ Microbiol. 2014 Apr;80(8):2516-25 [PMID: 24509927]
  10. Appl Environ Microbiol. 2011 Sep;77(17):6199-207 [PMID: 21764965]
  11. Lett Appl Microbiol. 2011 Apr;52(4):344-51 [PMID: 21244453]
  12. Lett Appl Microbiol. 2001 Mar;32(3):171-5 [PMID: 11264747]
  13. Food Microbiol. 2011 Sep;28(6):1238-41 [PMID: 21645826]
  14. Ann Agric Environ Med. 2015;22(3):429-32 [PMID: 26403108]
  15. Lett Appl Microbiol. 2010 Jan;50(1):82-8 [PMID: 19895420]
  16. Food Chem. 2015 Mar 1;170:470-83 [PMID: 25306373]
  17. Biosens Bioelectron. 2016 May 15;79:273-9 [PMID: 26710344]
  18. J Clin Microbiol. 2012 Jan;50(1):91-7 [PMID: 22031701]
  19. Mol Cell Probes. 2011 Aug;25(4):158-63 [PMID: 21513793]
  20. Parasitol Int. 2014 Dec;63(6):777-84 [PMID: 25038579]
  21. J Agric Food Chem. 2015 Jan 28;63(3):745-53 [PMID: 25539027]
  22. Food Microbiol. 2009 May;26(3):311-6 [PMID: 19269574]
  23. Appl Environ Microbiol. 2006 Jun;72(6):3896-900 [PMID: 16751494]

Word Cloud

Created with Highcharts 10.0.0detectionO157:H7samplefoodLAMP-LFAmethodisothermalflowrapidrealbeeffilteredLAMPreactionsensitiveusedRapidlownumberpathogenicbacteriadifficultstudytestedfilter-basedloop-mediatedamplification-lateralimmunoassaypathogensinoculated25 ghomogenized045 μmcellulosenitratefilterconcentratedrecoveredDNAextractedanalyzedLFAperformedhybridizationdigoxygenin-labeledampliconbiotinylatedprobesensitivity100 timesunfilteredtotaltimepreparationconfirmationwithin3 hresultssuggestcansystemspoint-of-caretestingDevelopmentfiltration-basedEscherichiacoliFiltrationLateralassayLoop-mediatedamplification

Similar Articles

Cited By