Title
Genotyping based on thermal denaturation of amplification products identifies species of the Mycobacterium tuberculosis complex
11627/517011627/5170
Author
Domínguez-Zepahua, Mariel Idalid
Hernández Arteaga, Luisa Eugenia del Socorro
López Revilla, Rubén Hipólito
Abstract
"Purpose. To develop a fast and inexpensive genotyping assay to identify the Mycobacterium tuberculosis complex (MTC) species most prevalent in human tuberculosis (TB), based on the thermal denaturation profiles of PCR products from mycobacterial 16S rDNA and three MTC genomic regions of difference (RD).
Methodology. Genotypes were determined by the presence and thermal denaturation profiles of the amplicons generated in the 'preliminary' PCR mixture (16S rDNA), followed by those of the simultaneous D1 (RD9+, RD1-) and D2 (RD4+, RD4-) PCR mixtures. The 16S rDNA profile identifies the genus Mycobacterium; the absence of any additional RD profile identifies Mycobacterium non-tuberculous (MNT) strains; additional RD4+ and RD9+ profiles without RD1- identify M. tuberculosis; an additional RD4+ profile per se identifies M. africanum; an additional RD4- profile per se identifies Mycobaterium bovis; additional RD1- and RD4- profiles identify M. bovis BCG.
Results. Genotypes of a panel with 44 mycobacterial strains coincided in 16 MB and five non-MTC strains; in the remaining 23 MTC strains, 17 MTB and five MA concordant genotypes and one discordant MB genotype were resolved. The genotypes of 13 human and bovine MTC isolates coincided in all four MB and eight of the nine MTB isolates.
Conclusion. Sensitivity, specificity and positive and negative predictive values of the method are 100% for the genus Mycobacterium, which resolves MB, MTB and MA genotypes. Species/genotype agreement is 97.7% for the panel and 92.3% for the MTC isolates. This method may be advantageously used to identify the most prevalent MTC species in humans."
Publication date
2018Publication type
articleDOI
https://doi.org/10.1099/jmm.0.000816Knowledge area
MICROBIOLOGÍACollections
Publisher
Microbiology SocietyKeywords
Mycobacterium tuberculosisMycobacterium bovis
Mycobacterium africanum
Mycobacterium tuberculosis complex
EvaGreen
Thermal denaturation
View/ Open
Metadata
Show full item record
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Related items
Showing items related by title, author, creator and subject.
-
Metodo para la identificación de Mycobacterium tuberculosis, Mycobacterium tuberculosis complex y Mycobacterium bovis en una muestra biológica, y uso de oligonucleótidos específicos
López Revilla, Rubén Hipólito; Alvarez Maya, Ikuri (2017-04)"En la presente invención se describe y reclama un método in Vitro para la identificación específica de Mycobacterium tuberculosis, Mycobacterium bovis y Mycobacterium Complex, de manera simultánea mediante PCR multiplex ... -
Mycobacterium tuberculosis TLR2 agonists LprA, LM and Man-LAM induce notch1 and socs3 transcription
Marino Marmolejo, Erika Nahomy; Tornez Benitez, Citlalli; Bonifaz Peña, Vania Elvira; Guzmán Rodríguez, Mabel; López González, Hugo Enrique; Gutiérrez Ortega, Abel; Santos Martínez, Martha Leticia; Flores Valdez, Mario Alberto (2011-08)"Mycobacterium tuberculosis employs a number of strategies to subvert host signaling events, leading to its persistence within macrophages. Upon infection, Mycobacterium bovis BCG induce the expression of suppressor of ... -
Diagnóstico molecular de Mycobacterium tuberculosis, M. bovis y M. bovis BCG mediante amplificación por círculo rodante con sondas padlock
Martínez Castellanos, Alan Ytzeen (2014-07)"CONTEXTO. El diagnóstico de la tuberculosis (TB) zoonótica requiere métodos rápidos, confiables y accesibles que distingan Mycobacterium bovis en la población humana de alto riesgo, pues los métodos microbiológicos ...