Advanced search
1 file | 346.91 KB

Molecular subtyping of Salmonella Typhimurium with multiplex oligonucleotide ligation-PCR (MOL-PCR)

Author
Organization
Project
Bioinformatics: from nucleotids to networks (N2N)
Abstract
A multiplex oligonucleotide ligation-PCR (MOL-PCR) assay is a valuable high-throughput technique for the detection of bacteria and viruses, for characterization of pathogens and for diagnosis of genetic diseases, as it allows one to combine different types of molecular markers in a high-throughput multiplex assay. A MOL-PCR assay starts with a multiplex oligonucleotide ligation reaction for detection of the molecular marker, followed by a singleplex PCR for signal amplification and analysis of the MOL-PCR products on a Luminex platform. This last step occurs through a liquid bead suspension array in which the MOL-PCR products are hybridized to MagPlex-TAG beads. In this chapter, we describe the complete procedure for a MOL-PCR assay for subtyping of Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) and its monophasic variant S. 1,4[5],12:i:- from DNA isolation through heat lysis up to data interpretation through a Gödel Prime Product. The subtyping assay consists of 50 discriminative molecular markers and two internal positive control markers divided over three MOL-PCR assays.
Keywords
MOL-PCR, Salmonella Typhimurium, Subtyping, Luminex, Bead suspension array, Gödel Prime Product, High-throughput assay

Downloads

  • (...).pdf
    • full text
    • |
    • UGent only
    • |
    • PDF
    • |
    • 346.91 KB

Citation

Please use this url to cite or link to this publication:

Chicago
Wuyts, Véronique, Wesley Mattheus, Nancy HC Roosens, Kathleen Marchal, Sophie Bertrand, and Sigrid CJ De Keersmaecker. 2017. “Molecular Subtyping of Salmonella Typhimurium with Multiplex Oligonucleotide ligation-PCR (MOL-PCR).” In Diagnostic Bacteriology : Methods and Protocols, ed. Kimberly A Bishop-Lilly, 1616:39–69. New York, NY, USA: Springer Humana Press.
APA
Wuyts, V., Mattheus, W., Roosens, N. H., Marchal, K., Bertrand, S., & De Keersmaecker, S. C. (2017). Molecular subtyping of Salmonella Typhimurium with multiplex oligonucleotide ligation-PCR (MOL-PCR). In K. A. Bishop-Lilly (Ed.), Diagnostic bacteriology : methods and protocols (Vol. 1616, pp. 39–69). New York, NY, USA: Springer Humana Press.
Vancouver
1.
Wuyts V, Mattheus W, Roosens NH, Marchal K, Bertrand S, De Keersmaecker SC. Molecular subtyping of Salmonella Typhimurium with multiplex oligonucleotide ligation-PCR (MOL-PCR). In: Bishop-Lilly KA, editor. Diagnostic bacteriology : methods and protocols. New York, NY, USA: Springer Humana Press; 2017. p. 39–69.
MLA
Wuyts, Véronique, Wesley Mattheus, Nancy HC Roosens, et al. “Molecular Subtyping of Salmonella Typhimurium with Multiplex Oligonucleotide ligation-PCR (MOL-PCR).” Diagnostic Bacteriology : Methods and Protocols. Ed. Kimberly A Bishop-Lilly. Vol. 1616. New York, NY, USA: Springer Humana Press, 2017. 39–69. Print.
@incollection{8546637,
  abstract     = {A multiplex oligonucleotide ligation-PCR (MOL-PCR) assay is a valuable high-throughput technique for the detection of bacteria and viruses, for characterization of pathogens and for diagnosis of genetic diseases, as it allows one to combine different types of molecular markers in a high-throughput multiplex assay. A MOL-PCR assay starts with a multiplex oligonucleotide ligation reaction for detection of the molecular marker, followed by a singleplex PCR for signal amplification and analysis of the MOL-PCR products on a Luminex platform. This last step occurs through a liquid bead suspension array in which the MOL-PCR products are hybridized to MagPlex-TAG beads.
In this chapter, we describe the complete procedure for a MOL-PCR assay for subtyping of Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) and its monophasic variant S. 1,4[5],12:i:- from DNA isolation through heat lysis up to data interpretation through a G{\"o}del Prime Product. The subtyping assay consists of 50 discriminative molecular markers and two internal positive control markers divided over three MOL-PCR assays.},
  author       = {Wuyts, V{\'e}ronique and Mattheus, Wesley and Roosens, Nancy HC and Marchal, Kathleen and Bertrand, Sophie and De Keersmaecker, Sigrid CJ},
  booktitle    = {Diagnostic bacteriology : methods and protocols},
  editor       = {Bishop-Lilly, Kimberly A},
  isbn         = {9781493970353},
  issn         = {1064-3745},
  keyword      = {MOL-PCR,Salmonella Typhimurium,Subtyping,Luminex,Bead suspension array,G{\"o}del Prime Product,High-throughput assay},
  language     = {eng},
  pages        = {39--69},
  publisher    = {Springer Humana Press},
  series       = {Methods in Molecular Biology},
  title        = {Molecular subtyping of Salmonella Typhimurium with multiplex oligonucleotide ligation-PCR (MOL-PCR)},
  url          = {http://dx.doi.org/10.1007/978-1-4939-7037-7\_3},
  volume       = {1616},
  year         = {2017},
}

Altmetric
View in Altmetric