A novel perspective on MOL-PCR optimization and MAGPIX analysis of in-house multiplex foodborne pathogens detection assay

Scientific Reports
Nikol ReslovaPetr Kralik

Abstract

Multiplex oligonucleotide ligation-PCR (MOL-PCR) is a rapid method for simultaneous detection of multiple molecular markers within a single reaction. MOL-PCR is increasingly employed in microbial detection assays, where its ability to facilitate identification and further characterization via simple analysis is of great benefit and significantly simplifies routine diagnostics. When adapted to microsphere suspension arrays on a MAGPIX reader, MOL-PCR has the potential to outperform standard nucleic acid-based diagnostic assays. This study represents the guideline towards in-house MOL-PCR assay optimization using the example of foodborne pathogens (bacteria and parasites) with an emphasis on the appropriate choice of crucial parameters. The optimized protocol focused on specific sequence detection utilizes the fluorescent reporter BODIPY-TMRX and self-coupled magnetic microspheres and allows for a smooth and brisk workflow which should serve as a guide for the development of MOL-PCR assays intended for pathogen detection.

References

Jan 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·F Barany
Sep 15, 1999·Journal of Immunological Methods·R T Carson, D A Vignali
Aug 3, 2001·International Journal of Food Microbiology·D R CallD P Chandler
Dec 10, 2003·Journal of Clinical Microbiology·Jeffrey HoorfarPatrick Fach
Aug 17, 2005·Clinica Chimica Acta; International Journal of Clinical Chemistry·Sherry A Dunbar
Apr 15, 2006·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·James W JacobsonJohn Kettman
May 2, 2008·Biosensors & Bioelectronics·Hyun-Joong KimHae-Yeong Kim
Jun 3, 2008·Nucleic Acids Research·Jason D Gans, Murray Wolinsky
Nov 15, 2008·BioTechniques·Shannon BruseLinda Brzustowicz
Dec 17, 2009·Journal of Microbiological Methods·Alina DeshpandeP Scott White
Feb 5, 2011·The American Journal of Tropical Medicine and Hygiene·Mami TaniuchiEric R Houpt
Nov 24, 2012·Critical Reviews in Microbiology·Saeed AkhtarAshfaque Hossain
Jul 25, 2015·Applied Microbiology and Biotechnology·Véronique WuytsSigrid C J De Keersmaecker
Feb 28, 2016·Journal of Immunological Methods·Jacqueline U McDonaldOmodele Ashiru
Sep 16, 2016·Frontiers in Cellular and Infection Microbiology·Travis A WoodsAlina Deshpande
Feb 10, 2017·Frontiers in Microbiology·Nikol ReslovaPetr Kralik
May 26, 2017·Sensors·Søren FjelstrupBirgitta Ruth Knudsen

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Methods Mentioned

BETA
PCR

Software Mentioned

Excel
Nucleotide BLAST
MOLigoDesigner
EliZyme
Plex Manager
MOL
ThermoNucleotideBlast
OligoAnalyzer
Bio
PCR

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