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Sharp RC, Naser ES, Alcedo KP, et al. Development of multiplex PCR and multi-color fluorescent in situ hybridization (. Gut Pathog. 2018;10:51doi: 10.1186/s13099-018-0278-1.
Sharp, R. C., Naser, E. S., Alcedo, K. P., Qasem, A., Abdelli, L. S., & Naser, S. A. (2018). Development of multiplex PCR and multi-color fluorescent in situ hybridization (. Gut pathogens, 1051. https://doi.org/10.1186/s13099-018-0278-1
Sharp, Robert C, et al. "Development of multiplex PCR and multi-color fluorescent in situ hybridization (." Gut pathogens vol. 10 (2018): 51. doi: https://doi.org/10.1186/s13099-018-0278-1
Sharp RC, Naser ES, Alcedo KP, Qasem A, Abdelli LS, Naser SA. Development of multiplex PCR and multi-color fluorescent in situ hybridization (. Gut Pathog. 2018 Dec 05;10:51. doi: 10.1186/s13099-018-0278-1. eCollection 2018. PMID: 30534203; PMCID: PMC6280354.
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Gut Pathog. 2018 Dec 05;10:51. doi: 10.1186/s13099-018-0278-1. eCollection 2018.

Development of multiplex PCR and multi-color fluorescent in situ hybridization (.

Gut pathogens

Robert C Sharp, Ebraheem S Naser, Karel P Alcedo, Ahmad Qasem, Latifa S Abdelli, Saleh A Naser

Affiliations

  1. Division of Molecular Microbiology, Burnett School of Biomedical Sciences, University of Central Florida College of Medicine, 4110 Libra Drive, Orlando, FL USA.

PMID: 30534203 PMCID: PMC6280354 DOI: 10.1186/s13099-018-0278-1

Abstract

BACKGROUND: Several pathogens have been debated to play a role in inflammatory bowel disease (IBD) including Crohn's disease (CD). None of these pathogens have been investigated together in same clinical samples. We developed a multiplex PCR and multi-color fluorescent in situ hybridization (

METHODS: The multiplex PCR is based on 1-h DNAzol

RESULTS: Following several attempts, the concentration of the oligonucleotide primers and DNA templates and the PCR annealing temperatures were optimized. Multiplex PCR analyses revealed excellent amplification signal in trials where a single primer set and combinations of two and three primers sets were tested against a mixture of DNA from three different bacteria or a mixture of three bacterial cultures mixed in one tube before DNA extraction. Slides with individual and mixtures of bacterial cultures and intestinal tissue sections from IBD patients were tested by

CONCLUSION: We developed a 4-h multiplex PCR protocol, which was validated by

Keywords: Crohn’s disease; MAP; Multiplex PCR; Ulcerative colitis; m-FISH

References

  1. Nature. 2011 Jun 15;474(7351):307-17 - PubMed
  2. Gut Microbes. 2012 Nov-Dec;3(6):544-55 - PubMed
  3. Curr Protoc Microbiol. 2009 Nov;Appendix 3:Appendix 3H - PubMed
  4. Am J Physiol Heart Circ Physiol. 2007 Aug;293(2):H1308-14 - PubMed
  5. Infect Immun. 2012 Nov;80(11):3768-75 - PubMed
  6. World J Gastroenterol. 2018 Feb 14;24(6):657-670 - PubMed
  7. Inflamm Bowel Dis. 2009 Jun;15(6):872-82 - PubMed
  8. Nature. 2012 Nov 1;491(7422):119-24 - PubMed
  9. Med Microbiol Immunol. 2015 Dec;204(6):681-92 - PubMed
  10. Gut Pathog. 2016 Mar 02;8:12 - PubMed
  11. EcoSal Plus. 2004 Dec;1(1):null - PubMed
  12. MedGenMed. 2004 Aug 10;6(3):14 - PubMed
  13. Curr Protoc Microbiol. 2009 Nov;Appendix 3:Appendix 3C - PubMed
  14. Gastroenterology. 2004 Aug;127(2):412-21 - PubMed
  15. Front Cell Infect Microbiol. 2018 Jan 25;8:11 - PubMed
  16. Scand J Gastroenterol. 2011 Jun;46(6):684-93 - PubMed
  17. Clin Microbiol Rev. 2007 Oct;20(4):593-621 - PubMed
  18. Front Cell Infect Microbiol. 2018 Jan 22;8:4 - PubMed
  19. Gastroenterol Hepatol (N Y). 2010 May;6(5):339-46 - PubMed
  20. Nat Rev Immunol. 2008 Jun;8(6):458-66 - PubMed
  21. Expert Rev Anti Infect Ther. 2017 Jul;15(7):637-643 - PubMed
  22. Clin Dev Immunol. 2013;2013:872632 - PubMed
  23. Lancet. 2004 Sep 18-24;364(9439):1039-44 - PubMed
  24. Infect Immun. 1999 Sep;67(9):4499-509 - PubMed
  25. Immunity. 2011 Mar 25;34(3):293-302 - PubMed
  26. Am J Gastroenterol. 2000 Apr;95(4):1094-5 - PubMed
  27. Front Microbiol. 2017 May 24;8:639 - PubMed
  28. Med Sci Monit. 2006 Apr;12(4):BR130-9 - PubMed
  29. PLoS One. 2015 Mar 13;10(3):e0120739 - PubMed
  30. World J Gastroenterol. 2014 Jun 21;20(23):7403-15 - PubMed
  31. Inflamm Bowel Dis. 2016 Jan;22(1):68-81 - PubMed
  32. Curr Opin Infect Dis. 2009 Jun;22(3):292-301 - PubMed
  33. Nature. 2015 Feb 19;518(7539):337-43 - PubMed

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