Search for Author, Title, Keyword
Rapid and Specific Detection of Escherichia coli O157:H7 in Ground Beef Using Immunomagnetic Separation Combined with Loop-Mediated Isothermal Amplification
 
More details
Hide details
Publication date: 2018-06-30
 
Pol. J. Food Nutr. Sci. 2018;68(2):115–123
 
KEYWORDS
ABSTRACT
Escherichia coli O157:H7 is well known for many foodborne outbreaks that lead to fatal infections in human being worldwide. The objective of this study was to develop a rapid and sensitive method for detection of EHEC O157:H7 from ground beef using a method that combined immunomagnetic separation (IMS) with loop-mediated isothermal amplification (LAMP). The EHEC O157:H7 cells were separated with Dynabeads coated with anti-EHEC O157:H7 after a short enrichment for 4 h. Then, EHEC O157:H7 was identified by LAMP assay for amplifying and detecting the rfbE gene, which is highly conserved in all EHEC O157:H7 strains and exhibits strain-specific gene expression. The LAMP method results analyzed with real time turbidity measurements showed a high specificity and sensitivity, with a positive detection rate of amplification of EHEC O157:H7 DNA diluted to a minimum equivalent concentration of 1.8 × 101 CFU/mL, which was 10 times more sensitive than the conventional PCR assay. The IMS followed with LAMP could capture and detect a bacterial concentration as low as 3×101 CFU/mL from the meat samples, which was close to the sensitivity of LAMP assay with pure culture. IMS combined with realistic LAMP method is a simple, rapid, highly specific gene amplification technology that is suitable for implementing as a screening assay in basic laboratory and field test for detecting food contamination.
 
REFERENCES (48)
1.
Aydin M., Herzig G.P., Jeong K.C., Dunigan S., Shah P., Ahn S., Rapid and sensitive detection of Escherichia coli O157:H7 in milk and ground beef using magnetic bead-based immunoassay coupled with tyramide signal amplification. J. Food Prot., 2014, 77, 100-105.
 
2.
Bapanpally C., Maganty G., Khan S., Kasra A., Morey A., Immunomagnetic separation and visual fluorescence detection of E. coli O157 using AOAC approved SAS Molecular Tests. J. AOAC Int., 2014, 97, 1073-1077.
 
3.
Bell B.P., Griffin P.M., Lozano P., Christie D.L., Kobayashi J.M., Tarr P.I., Predictors of hemolytic uremic syndrome in children during a large outbreak of Escherichia coli O157:H7 infections. Pediatrics, 1997, 100, E12.
 
4.
Bilge S.S., Vary J.C., Jr, Dowell S.F,, Tarr PI., Role of the Escherichia coli O157:H7 O side chain in adherence and analysis of an rfb locus. Infect. Immun., 1996, 64, 4795-4801.
 
5.
Buesa J.B., Collado P., López-Andújar P., Abu-Mallouh R., Rodríguez Díaz J., García Díaz A., Molecular epidemiology of calici viruses causing outbreaks and sporadic cases of acute gastroenteritis in Spain. J. Clin. Microbiol., 2002, 40, 2854-2859..
 
6.
CDC, Centers for Disease Control and Prevention, Importance of culture confirmation of Shiga toxin-producing Escherichia coli infection as illustrated by outbreaks of gastroenteritis— New York and North Carolina, 2005. MMWR Morb Mortal Wkly Rep., 2006, 55, 1042-1045.
 
7.
Chapman P.A., Ellin M., Ashton R., A comparison of immunomagnetic separation and culture, RevealTM 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., 2001, 32, 171-175.
 
8.
Chase-Topping M., Gally D., Low C., Matthews L., Woolhouse M., Super-shedding and the link between human infection and livestock carriage of Escherichia coli O157. Nat. Rev. Microbiol., 2008, 6, 904-912.
 
9.
Desmarchelier P.M., Bilge S.S., Fegan N., Mills L., Vary J.C.Jr., Tarr P.I., A PCR specific for Escherichia coli O157 based on the rfb locus encoding O157 lipopolysaccharide. J. Clin. Microbiol., 1998, 36, 1801-1804.
 
10.
Fan H., Long B., Wu X., Bai Y., Development of a Loop-Mediated Isothermal Amplification Assay for sensitive and rapid detection of Cronobacter sakazakii. Foodborne Pathog. Dis., 2012, 9, 1111-1118.
 
11.
Francois P., Tangomo M., Hibbs J., Bonetti E.J., Boehme C.C., Notomi T., Perkins M.D., Schrenzel J., Robustness of a loop-mediated isothermal amplification reaction for diagnostic applications. FEMS Immunol. Med. Microbiol., 2011, 62, 41–48.
 
12.
Fratamico P.M., Bagi L.K., Cray W.C. Jr., Narang N., Yan X., Medina M., Liu Y., Detection by multiplex real-time polymerase chain reaction assays and isolation of Shiga toxin-producing Escherichia coli serogroups O26, O45, O103, O111, O121, and O145 in ground beef. Foodborne Pathog. Dis., 2011, 8, 601–607.
 
13.
Gianviti A., Rosmini F., Caprioli A., Corona R., Matteucci MC., Principato F., Luzzi I., Rizzoni G., Haemolytic-uraemic syndrome in childhood: surveillance and case-control studies in Italy. Italian HUS Study Group. Pediatr. Nephrol., 1994, 8, 705–9.
 
14.
Gould L.H., Bopp C., Strockbine N., Atkinson R., Baselski V., Body B., Carey R., Crandall C., Hurd S., Kaplan R., Neill M., Shea S., Somsel P., Tobin-D'Angelo M., Griffin P.M., Gerner-Smidt P., Centers for Disease Control and Prevention (CDC), Recommendations for diagnosis of Shiga toxin–producing Escherichia coli infections by clinical laboratories. MMWR Recommend Rep., 2009, 58, 1–14.
 
15.
Han F., Ge B., Quantitative detection of Vibrio vulnificus in raw oysters by real-time loop-mediated isothermal amplification. Int. J. Food Microbiol., 2010, 142, 60–66.
 
16.
Hara-Kudo Y., Nemoto J., Ohtsuka K., Segawa Y., Takatori K., Kojima T., Ikedo M., Sensitive and rapid detection of Vero toxin-producing Escherichia coli using loop-mediated isothermal amplification. J. Med. Microbiol., 2007, 56, 398-406.
 
17.
Hara-Kudo Y., Konishi N., Ohtsuka K., Hiramatsu R, Tanaka H, Konuma HK, Takatori K., Detection of verotoxigenic Escherichia coli O157 and O26 in food by plating methods and LAMP method: A collaborative study. Int. J. Food Microbiol., 2008, 122, 156-161.
 
18.
Hill J., Beriwal S., Chandra I., Paul V.K., Kapil A., Singh T., Wadowsky R.M., Singh V., Goyal A., Jahnukainen T., Johnson J.R., Tarr P.I., Vats A., Loop-mediated isothermal amplification assay for rapid detection of common strains of Escherichia coli. J. Clin. Microbiol., 2008, 46, 2800–2804.
 
19.
Jeníková G., Pazlarová J., Demnerová K., Detection of Salmonella in food samples by the combination of immunomagnetic separation and PCR assay. Int. Microbiol., 2000, 3, 225–229.
 
20.
Jeon S.J., Elzo M., Dilorenzo N., Lamb G.C., Jeong K.C., Evaluation of animal genetic and physiological factors that affect the prevalence of Escherichia coli O157 in cattle. PLoS One, 2013, 8, e55728.
 
21.
Jiang R., Long B., Zeng G., Wang D., Fan H., Wu X., Loop-mediated isothermal amplification for detecting enterohemorrhagic Escherichia coli O157:H7: a comparison with PCR. Nan Fang Yi Ke Da Xue Xue Bao., 2012, 32, 1026-1030.
 
22.
Karch H., Tarr P.I., Bielaszewska M., Enterohaemorrhagic Escherichia coli in human medicine. Int. J. Med. Microbiol., 2005, 295, 405-418.
 
23.
Kou X., Wu Q., Zhang J., Fan H., Development and application of a loop-mediated isothermal amplification assay on rapid and sensitive detection of rotavirus in fecal samples and artificially seeded oysters. Food Contr., 2014, 41, 151-157.
 
24.
Mckee M.L., O’Brien A.D., Investigation of enterohemorrhagic Escherichia coli O157:H7 adherence characteristics and invasion potential reveals a new attachment pattern shared by intestinal E.coli. Infect Immun., 1995, 63, 2070-2074.
 
25.
Mori Y., Nagamine K., Tomita N., Notomi T., Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem. Biophys. Res. Commun., 2001, 289, 150–154.
 
26.
Mori Y., Notomi T., Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases. J. Infect. Chemother., 2009, 15, 62–69.
 
27.
Nagamine K., Watanage K., Ohtsuka K., Hase T., Notomi T., Loop-mediated isothermal amplification reaction using a nondenatured template. Clin. Chem., 2001, 47, 1742-1743.
 
28.
Nagamine K., Hase T., Notomi T., Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol. Cell. Probes, 2002, 16, 223–229.
 
29.
Nemoto J., Sugawara C., Akahane K., Hashimoto K., Kojima T., Ikedo M., Konuma H., Hara-Kudo Y., Rapid and specific detection of the thermostable direct hemolysin gene in Vibrio parahaemolyticus by loop-mediated isothermal amplification. J. Food Prot., 2009, 72, 748–754.
 
30.
Notomi T., Okayama H., Masubuchi H., Yonekawa T., Watanabe K., Amino N., Hase T., Loop-mediated isothermal amplification of DNA. Nucleic Acids Res., 2000, 28, E63.
 
31.
Onoue Y., Konuma H., Nakagawa H., Hara-Kudo Y., Fujita T., Kumagai S., Collaborative evaluation of detection methods for Escherichia coli O157:H7 from radish sprouts and ground beef. Int. J. Food Microbiol., 1999, 46, 27-36.
 
32.
Orskov F., Orskov I., Escherichia coli serotyping and disease in man and animals. Can. J. Microbiol., 1992, 38, 699-704.
 
33.
Pruimboom-Brees I.M., Morgan T.W., Ackermann M.R., Nystrom E.D., Samuel J.E., Cornick N.A., Moon H.W., Cattle lack vascular receptors for Escherichia coli O157:H7 Shiga toxins. Proc. Natl. Acad. Sci. USA., 2000, 97, 10325-10329.
 
34.
Ravan H.,, Amandadi M., Sanadgol N., A highly specific and sensitive loop-mediated isothermal amplification method for the detection of Escherichia coli O157:H7. Microb. Pathogen., 2016, 91, 161-165.
 
35.
Saito R., Misawa Y., Moriya K., Koike K., Ubukata K., Okamura N., Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of Mycoplasma pneumonia. J. Med. Microbiol., 2005, 54, 1037-1041.
 
36.
Sattabongkot J., Tsuboi T., Han E.T., Bantuchai S., Buates S., Loop-mediated isothermal amplification assay for rapid diagnosis of malaria infections in an area of endemicity in Thailand. J. Clin. Microbiol., 2014, 52, 1471–1477.
 
37.
Sen K., Ashbolt N.J., Environmental Microbiology: Current Technology and Water Application. 2011, Caister Academic Press, Norfolk, UK, ISBN; 978(1): 904455-70-7.
 
38.
Sharma V.K., Dean-Nystrom E.A., Detection of enterohemorrhagic Escherichia coli O157:H7 by using a multiplex real-time PCR assay for genes encoding intimin and Shiga toxins. Vet. Microbiol., 2003, 93, 247-260.
 
39.
Taguchi T., Arakaki A., Takeyama H., Haraguchi S., Yoshino M., Kaneko M., Detection of Cryptosporidium parvum oocysts using a microfluidic device equipped with the SUS micromesh and FITC. Biotechnol. Bioeng., 2007, 96, 272-280.
 
40.
Wang F., Jiang L., Ge B., Loop-mediated isothermal amplification assays for detecting shiga toxin-producing Escherichia coli in ground beef and human stools. J. Clin. Microbiol., 2012, 50, 91-97.
 
41.
Wang Y., Wang Y., Luo L., Liu D., Luo X., Xu Y., Hu S., Niu L., Xu J., Ye C., Rapid and sensitive detection of Shigella spp. and Salmonella spp. by Multiple Endonuclease Restriction Real-Time Loop-Mediated Isothermal Amplification Technique. Front. Microbiol., 2015, 6, 1400.
 
42.
Wani S.A., Hussain I., Nabi A., Fayaz I., Nishikawa Y., Variants of eae and stx genes of atypical enteropathogenic Escherichia coli and non-O157 Shiga toxin-producing Escherichia coli from calves. Lett. Appl. Microbiol., 2007, 45, 610-615.
 
43.
Wolf S., Williamson W.M., Hewitt J., Rivera-Aban M., Lin S., Ball A., Scholes P., Greening GE., Sensitive multiplex real—time reverse transcription-PCR assay for the detection of human and animal noroviruses in clinical and environmental samples. Appl. Environ. Microbiol., 2007, 73, 5464—5470.
 
44.
Wright D.J., Chapman P.A., Siddons C.A., Immunomagnetic separation as a sensitive method for isolating E. coli O157 from food samples. Epidemiol. Infect., 1994, 113, 31-39.
 
45.
Xiong Y., Wang P., Lan R., Ye C., Wang H., Ren J., Jing H., Wang Y., Zhou Z., Bai X., Cui Z., Luo X., Zhao A., Wang Y., Zhang S., Sun H., Wang L., Xu J., A novel Escherichia coli O157:H7 clone causing a major hemolytic uremic syndrome outbreak in China. PLoS One, 2012, 7, e36144.
 
46.
Yamazaki W., Ishibashi M., Kawahara R., Inoue K., Development of a loop-mediated isothermal amplification assay for sensitive and rapid detection of Vibrio parahaemolyticus. BMC Microbiol., 2008, 8, 163.
 
47.
Zeng J., Wei H., Zhang L., Liu X., Zhang H., Cheng J., Ma D., Zhang X., Fu P., Liu L., Rapid detection of Vibrio parahaemolyticus in raw oysters using immunomagnetic separation combined with loop-mediated isothermal amplification. Int. J. Food Microbiol., 2014, 174, 123-128.
 
48.
Zhao X., Li Y., Wang L., You L., Xu Z., Li L., He X., Liu Y., Wang J., Yang L., Development and application of a loop-mediated isothermal amplification method on rapid detection Escherichia coli O157 strains from food samples. Mol. Biol. Rep., 2010, 37, 2183-2188.
 
 
CITATIONS (3):
1.
An evaluation of immunomagnetic separation-real-time PCR (IMS-RTiPCR) combined assay for rapid and specific detection of Escherichia coli O157:H7 in raw milk and ground beef
TABAN MERCANOGLU, Sait AYTAC
Food Science and Technology
 
2.
Immunomagnetic separation: An effective pretreatment technology for isolation and enrichment in food microorganisms detection
Zhouli Wang, Rui Cai, Zhenpeng Gao, Yahong Yuan, Tianli Yue
Comprehensive Reviews in Food Science and Food Safety
 
3.
Newly isolated Coliphages for Bio-controlling Multidrug Resistant Escherichia coli strains
Mostafa Elbahnasawy, ElSayed ElSayed, Mohamed Azzam
Environmental Nanotechnology, Monitoring & Management
 
eISSN:2083-6007
ISSN:1230-0322