title: Quick and cheap MIRU-VNTR typing of Mycobacterium tuberculosis species complex using duplex PCR.
abstract: While minisatellites are usually typed using capillary sequencers or qiaplex systems in developed countries, many low-resource regions cannot afford it. We propose an optimized agarose gel electrophoresis method to genotype Mycobacterium tuberculosis species complex minisatellites in their standardized format (24 MIRU-VNTR). It is based on duplex PCRs combining VNTR loci harboring distinct amplicon sizes whatever the repetition number of each locus. This method performs well both on DNA extracts of good quality and on thermolysates while reducing workload and reagents costs.
authors.affiliation: Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), P.O. Box# 577, Jhang Road, Faisalabad, Pakistan; Pakistan Institute of Engineering and Applied Sciences (PIEAS), Nilore, Islamabad, Pakistan.
authors.forename: Memona
authors.initials: M
authors.lastname: Yasmin
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris-Sud, Université Paris-Saclay, 91198, Gif-sur-Yvette Cedex, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Le Moullec
authors.affiliation: Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), P.O. Box# 577, Jhang Road, Faisalabad, Pakistan.
authors.forename: Rubina Tabassum
authors.initials: RT
authors.lastname: Siddiqui
authors.affiliation: National Tuberculosis Reference Laboratory, National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands.
authors.forename: Jessica
authors.initials: J
authors.lastname: De Beer
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris-Sud, Université Paris-Saclay, 91198, Gif-sur-Yvette Cedex, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Sola
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris-Sud, Université Paris-Saclay, 91198, Gif-sur-Yvette Cedex, France. Electronic address: guislaine.refregier@u-psud.fr.
authors.forename: Guislaine
authors.initials: G
authors.lastname: Refrégier
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics; D004587:Electrophoresis, Agar Gel / Q000379:methods; D060005:Genotyping Techniques / Q000379:methods; D006801:Humans; D018598:Minisatellite Repeats / Q000235:genetics; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000235:genetics*; D014376:Tuberculosis / Q000382:microbiology
chemical_list: D004269:DNA, Bacterial
keywords: Genotyping; MLVA; Multiplexing