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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
medicine
"Spoligoriftyping," a dual-priming-oligonucleotide-based direct-hybridization assay for tuberculosis control with a multianalyte microbead-based hybridization system
Journal of Clinical Microbiology, Volume 50, No. 10, Year 2012
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Description
We developed "spoligoriftyping," a 53-plex assay based on two preexisting methods, the spoligotyping and "rifoligotyping" assays, by combining them into a single assay. Spoligoriftyping allows simultaneous spoligotyping (i.e., clustered regularly interspaced short palindromic repeat [CRISPR]-based genotyping) and characterization of the main rifampin drug resistance mutations on the rpoB hot spot region in a few hours. This test partly uses the dual-priming-oligonucleotide (DPO) principle, which allows simultaneous efficient amplifications of rpoB and the CRISPR locus in the same sample. We tested this method on a set of 114 previously phenotypically and genotypically characterized multidrug-resistant (MDR) Mycobacterium tuberculosis or drugsusceptible M. tuberculosis DNA extracted from clinical isolates obtained from patients from Bulgaria, Nigeria, and Germany. We showed that our method is 100% concordant with rpoB sequencing results and 99.95% (3,911/3,913 spoligotype data points) correlated with classical spoligotyping results. The sensitivity and specificity of our assay were 99 and 100%, respectively, compared to those of phenotypic drug susceptibility testing. Such assays pave the way to the implementation of locally and specifically adapted methods of performing in a single tube both drug resistance mutation detection and genotyping in a few hours. Copyright © 2012, American Society for Microbiology. All Rights Reserved.
Available Materials
https://efashare.b-cdn.net/share/pmc/articles/PMC3457427/bin/supp_50_10_3172__index.html
https://efashare.b-cdn.net/share/pmc/articles/PMC3457427/bin/JCM.00976-12_zjm999091929so1.pdf
https://efashare.b-cdn.net/share/pmc/articles/PMC3457427/bin/JCM.00976-12_zjm999091929so2.xls
Authors & Co-Authors
Gomgnimbou, Michel Kiréopori
France, Paris
Cnrs Centre National de la Recherche Scientifique
Burkina Faso, Ouagadougou
Centre Muraz
Abadia, Edgar
Venezuela, Caracas
Instituto Venezolano de Investigaciones Cientificas, Centro de Microbiologia y Biologia Celular
Zhang, Jian
France, Paris
Cnrs Centre National de la Recherche Scientifique
Refrégier, Guislaine
France, Paris
Cnrs Centre National de la Recherche Scientifique
Panaiotov, S.
Bulgaria, Sofia
National Center of Infectious and Parasitic Diseases Bulgaria
Bachiyska, Elizabeta V.
Bulgaria, Sofia
National Center of Infectious and Parasitic Diseases Bulgaria
Sola, Christophe Noel
France, Paris
Cnrs Centre National de la Recherche Scientifique
Statistics
Citations: 33
Authors: 7
Affiliations: 4
Identifiers
Doi:
10.1128/JCM.00976-12
ISSN:
00951137
e-ISSN:
1098660X
Research Areas
Cancer
Genetics And Genomics
Study Locations
Nigeria