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Guidelines for Optimisation of a Multiplex Oligonucleotide Ligation-PCR for Characterisation of Microbial Pathogens in a Microsphere Suspension Array
Joint Authors
De Keersmaecker, Sigrid C. J.
Wuyts, Véronique
Bertrand, Sophie
Marchal, Kathleen
Roosens, Nancy H. C.
Source
Issue
Vol. 2015, Issue 2015 (31 Dec. 2015), pp.1-10, 10 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2015-02-03
Country of Publication
Egypt
No. of Pages
10
Main Subjects
Abstract EN
With multiplex oligonucleotide ligation-PCR (MOL-PCR) different molecular markers can be simultaneously analysed in a single assay and high levels of multiplexing can be achieved in high-throughput format.
As such, MOL-PCR is a convenient solution for microbial detection and identification assays where many markers should be analysed, including for routine further characterisation of an identified microbial pathogenic isolate.
For an assay aimed at routine use, optimisation in terms of differentiation between positive and negative results and of cost and effort is indispensable.
As MOL-PCR includes a multiplex ligation step, followed by a singleplex PCR and analysis with microspheres on a Luminex device, several parameters are accessible for optimisation.
Although MOL-PCR performance may be influenced by the markers used in the assay and the targeted bacterial species, evaluation of the method of DNA isolation, the probe concentration, the amount of microspheres, and the concentration of reporter dye is advisable in the development of any MOL-PCR assay.
Therefore, we here describe our observations made during the optimisation of a 20-plex MOL-PCR assay for subtyping of Salmonella Typhimurium with the aim to provide a possible workflow as guidance for the development and optimisation of a MOL-PCR assay for the characterisation of other microbial pathogens.
American Psychological Association (APA)
Wuyts, Véronique& Roosens, Nancy H. C.& Bertrand, Sophie& Marchal, Kathleen& De Keersmaecker, Sigrid C. J.. 2015. Guidelines for Optimisation of a Multiplex Oligonucleotide Ligation-PCR for Characterisation of Microbial Pathogens in a Microsphere Suspension Array. BioMed Research International،Vol. 2015, no. 2015, pp.1-10.
https://search.emarefa.net/detail/BIM-1056731
Modern Language Association (MLA)
Wuyts, Véronique…[et al.]. Guidelines for Optimisation of a Multiplex Oligonucleotide Ligation-PCR for Characterisation of Microbial Pathogens in a Microsphere Suspension Array. BioMed Research International No. 2015 (2015), pp.1-10.
https://search.emarefa.net/detail/BIM-1056731
American Medical Association (AMA)
Wuyts, Véronique& Roosens, Nancy H. C.& Bertrand, Sophie& Marchal, Kathleen& De Keersmaecker, Sigrid C. J.. Guidelines for Optimisation of a Multiplex Oligonucleotide Ligation-PCR for Characterisation of Microbial Pathogens in a Microsphere Suspension Array. BioMed Research International. 2015. Vol. 2015, no. 2015, pp.1-10.
https://search.emarefa.net/detail/BIM-1056731
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1056731