1. bookVolume 26 (2018): Edition 2 (April 2018)
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Magazine
eISSN
2284-5623
Première parution
08 Aug 2013
Périodicité
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Anglais
access type Accès libre

xTAG Luminex multiplex assay for rapid screening of verocytotoxin-producing Escherichia coli strains

Publié en ligne: 17 May 2018
Volume & Edition: Volume 26 (2018) - Edition 2 (April 2018)
Pages: 211 - 219
Reçu: 29 Jan 2018
Accepté: 05 Apr 2018
Détails du magazine
License
Format
Magazine
eISSN
2284-5623
Première parution
08 Aug 2013
Périodicité
4 fois par an
Langues
Anglais
Abstract

The O26 verocytotoxin-producing Escherichia coli (VTEC)-associated outbreak of hemolytic uremic syndrome (HUS) cases in Romania during 2016 showed the need to improve the current methodology of non-O157 VTEC detection and surveillance. An in-house assay based on xTAG Luminex technology was optimized to identify seven of the most relevant diarrheagenic E.coli serogroups (O-specific wzx genes), two convenient VTEC virulence markers (eaeA and ehxA genes), and a species-specific control gene (uidA). Twenty-nine strains previously characterized in terms of serogroup and virulence genes were tested with the optimized protocol and the results were as expected. The ratio of sample signal to background varied from 66.7 (ehxA) to 7.6 (uidA) for positive samples, with a cut-off of 3. Sensitivity varied depending on the target to be amplified from approximately 102 genomic copies to approximately 104 genomic copies per reaction, respectively. The current approach seems an affordable alternative to commercially available assays that can be further exploited to improve existing autochthonous strategies to prevent future VTEC outbreaks.

Keywords

1. Etcheverría AI, Padola NL. Shiga toxin-producing Escherichia coli: factors involved in virulence and cattle colonization. Virulence. 2013;4:366-72. DOI: 10.4161/viru.2464210.4161/viru.24642371412823624795Ouvrir le DOISearch in Google Scholar

2. Croxen MA, Law RJ, Scholz R, Keeney KM, Wlodarska M, Finlay BB. Recent advances in understanding enteric pathogenic Escherichia coli. Clin Microbiol Rev. 2013;26(4):822-80. DOI: 10.1128/CMR.00022-1310.1128/CMR.00022-13381123324092857Ouvrir le DOISearch in Google Scholar

3. Kaper JB, Nataro JP, Mobley HL. Pathogenic Escherichia coli. Nat Rev Microbiol. 2004;2(2):123-40. DOI: 10.1038/nrmicro81810.1038/nrmicro81815040260Ouvrir le DOISearch in Google Scholar

4. Scheutz F, Strockbine NA. Escherichia. In: Garrity GM, Brenner DJ, Krieg NR, Staley JT, editors. Bergey’s manual of systematic bacteriology. New York: Springer; 2005. P. 607-624.Search in Google Scholar

5. Fratamico PM, DebRoy C, Liu Y, Needleman DS, Baranzoni GM, Feng P. Advances in molecular serotyping and subtyping of Escherichia coli. Front Microbiol. 2016;7:644. DOI: 10.3389/fmicb.2016.0064410.3389/fmicb.2016.00644485340327199968Ouvrir le DOISearch in Google Scholar

6. Iguchi A, Iyoda S, Kikuchi T, Ogura Y, Katsura K, Ohnishi M et al. A complete view of the genetic diversity of the Escherichia coli O-antigen biosynthesis gene cluster. DNA Res 2015;22(1):101-7. DOI: 10.1093/dnares/dsu04310.1093/dnares/dsu043437998125428893Ouvrir le DOISearch in Google Scholar

7. Johnson KE, Thorpe CM, Sears CL. The emergingclinical importance of non-O157 Shiga toxin-producing Escherichia coli. Clin Infect Dis 2006;43:1587-95.DOI: 10.1086/50957310.1086/50957317109294Ouvrir le DOISearch in Google Scholar

8. Muniesa M, Hammerl JA, Hertwig S, Appel B, Brüssow H. Shiga toxin-producing Escherichia coli O104:H4: a new challenge for microbiology. Appl Environ Microbiol 2012;78(12):4065-73. DOI: 10.1128/AEM.00217-1210.1128/AEM.00217-12337053422504816Search in Google Scholar

9. Peron E, Zaharia A, Zota LC, Severi E, Mårdh O, Usein C, et al. Early findings in outbreak of haemolytic uraemic yndrome among young children caused by Shiga toxin-producing Escherichia coli, Romania, January to February 2016. Euro Surveill 2016;21(11):30170. DOI: 10.2807/1560-7917.ES.2016.21.11.3017010.2807/1560-7917.ES.2016.21.11.3017027020906Ouvrir le DOISearch in Google Scholar

10. Usein CR, Ciontea AS, Militaru CM, Condei M, Dinu S, Oprea M, et al. Molecular characterisation of human Shiga toxin-producing Escherichia coli O26 strains: results of an outbreak investigation, Romania, February to August 2016. Euro Surveill 2017;22(47):17-00148. DOI: 10.2807/1560-7917.ES.2017.22.47.17-0014810.2807/1560-7917.ES.2017.22.47.17-00148571066029183554Search in Google Scholar

11. Luminex xMAP® Cookbook. A collection of methods and protocols for developing multiplex assays with xMAP Technology. 2rd Edition. Available at http://info.luminexcorp.com/en-us/download-the-xmap-cookbook Accessed November 14, 2016.Search in Google Scholar

12. DebRoy C, Roberts E, Valadez AM, Dudley EG, Cutter CN. Detection of Shiga toxin-producing Escherichia coli O26, O45, O103, O111, O113, O121, O145, and O157 serogroups by multiplex polymerase chain reaction of the wzx gene of the O-antigen gene cluster. Foodborne Pathog Dis 2011;8(5):651-2. DOI: 10.1089/fpd.2010.076910.1089/fpd.2010.076921548768Ouvrir le DOISearch in Google Scholar

13. Leszczyńska B, Ziółkowska H, Podsiadły E, Szych J, Rastawicki W, Demkow U, et al. Diagnostic value of serological tests against verotoxigenic Escherichia coli in hemolytic uremic syndrome in children. Adv Clin Exp Med. 2015;24(6):1031-6. DOI: 10.17219/acem/4193110.17219/acem/4193126771976Ouvrir le DOISearch in Google Scholar

14. Rice T, Quinn N, Sleator RD, Lucey B. Changing diagnostic methods and increased detection of verotoxigenic Escherichia coli, Ireland. Emerg Infect Dis. 2016;22(9):1656-7. DOI: 10.3201/eid2209.16047710.3201/eid2209.160477499436527322897Ouvrir le DOISearch in Google Scholar

15. European Centre for Disease Prevention and Control. Annual Epidemiological Report 2016 - Shigatoxin/ verocytotoxin-producing Escherichia coli infection. Available at http://ecdc.europa.eu/en/healthtopics/stec/Pages/Annual-epidemiological-report-2016.aspx Accessed December 20, 2017.Search in Google Scholar

16. Wessels E, Rusman LG, van Bussel MJ, Claas EC. Added value of multiplex Luminex Gastrointestinal Pathogen Panel (xTAG® GPP) testing in the diagnosis of infectious gastroenteritis. Clin Microbiol Infect 2014;20(3):O182-O187. DOI: 10.1111/1469-0691.1236410.1111/1469-0691.1236424131399Ouvrir le DOISearch in Google Scholar

17. Carter JM1 Lin A, Clotilde L, Lesho M. Rapid, multiplexed characterization of Shiga toxin-producing Escherichia coli (STEC) Isolates Using Suspension Array Technology. Front Microbiol 2016;7:439.Search in Google Scholar

18. Jiang L, Ren H, Zhou H, Qin T, Chen Y. Simultaneous detection of nine key bacterial respiratory pathogens using Luminex xTAG(®) technology. Int J Environ Res Public Health 2017;14(3):E223. DOI: 10.3390/ijerph1403022310.3390/ijerph14030223536905928241513Ouvrir le DOISearch in Google Scholar

19. Lin A, Nguyen L, Lee T, Clotilde LM, Kase JA, Son I, Carter JM, Lauzon CR. Rapid O serogroup identification of the ten most clinically relevant STECs by Luminex microbead-based suspension array. J Microbiol Methods 2011;87(1):105-10. DOI: 10.1016/j.mimet.2011.07.01910.1016/j.mimet.2011.07.01921835211Search in Google Scholar

20. Clotilde LM, Bernard C 4th, Hartman GL, Lau DK, Carter JM. Microbead-based immunoassay for simultaneous detection of Shiga toxins and isolation of Escherichia coli O157 in foods. J Food Prot 2011;74(3):373-9. DOI: 10.4315/0362-028X.JFP-10-34410.4315/0362-028X.JFP-10-34421375872Ouvrir le DOISearch in Google Scholar

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