Distribución y frecuencia de potenciales mutaciones asociadas con la resistencia a rifampicina en el gen rpoB de Mycobacterium tuberculosis detectadas mediante un método molecular automatizado

Publicado

2023-01-01

Distribution and frequency of potential mutations associated with rifampicin resistance in the rpoB gene of Mycobacterium tuberculosis detected using a molecular automated method

Distribución y frecuencia de potenciales mutaciones asociadas con la resistencia a rifampicina en el gen rpoB de Mycobacterium tuberculosis detectadas mediante un método molecular automatizado

Palabras clave:

Colombia, Mutation, Polymerase Chain Reaction, Tuberculosis, Rifampicin (en)
Colombia, Mutación, Reacción en cadena de la polimerasa, Tuberculosis, Rifampicina (es)

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Autores/as

Adrián Peñata Hospital San Vicente Fundación - Laboratorio de Biología Molecular e Inmunología de Trasplantes - Medellín - Colombia. https://orcid.org/0000-0002-7515-6718
Camilo Pérez Universidad de Antioquia - Escuela de Microbiología - Medellín - Colombia. https://orcid.org/0000-0001-7882-2862
Tatiana Castaño Hospital San Vicente Fundación - Laboratorio de Biología Molecular e Inmunología de Trasplantes - Medellín - Colombia. https://orcid.org/0000-0002-3067-2436
Julián Bustamante Hospital San Vicente Fundación - Laboratorio de Biología Molecular e Inmunología de Trasplantes - Medellín - Colombia. https://orcid.org/0000-0003-1738-0997
Sigifredo Ospina Universidad de Antioquia - Facultad de Medicina - Instituto de Investigaciones Médicas - Medellín - Colombia. https://orcid.org/0000-0002-1241-4177

Resumen (en)
Resumen (es)

Introduction: Tuberculosis continues to be a public health problem that is now aggravated by drug-resistant Mycobacterium tuberculosis. More than 95% of rifampicin (RIF)-resistant strains of M. tuberculosis have mutations in a region of the rpoB gene. Xpert^TM MTB/RIF is a molecular biology system that allows identifying mutations in the RIF-resistance determining region of this gen using 5 probes (A, B, C, D, E) that make up the rpoB gene sequences.

Objective: To describe the distribution and frequency of potential mutations associated with RIF resistance in the rpoB gene of M. tuberculosis detected in pulmonary and extrapulmonary samples using the Xpert® MTB/RIF method.

Materials and methods: Retrospective study in which 66 samples positive for RIF-resistant M. tuberculosis, processed using the GeneXpert MTB/RIF system between January 2011 and July 2019 at a university hospital in Medellín, Colombia, were analyzed. According to the Dx System software of the GenXpert instrument, a potential mutation and RIF resistance were established if the probes did not bind to their natural complementary beacon or if there was a delay in binding (delta C_T>4) in relation the other probes due to the presence of an abnormal beacon. Data were analyzed using descriptive statistics.

Results: Of the 66 samples (48 pulmonary and 18 extrapulmonary), 63.64% were from men and participants’ mean age was 39.60 ± 17.69 years. The frequency and distribution of mutations was as follows: probe E: 38 mutations (57.58%), B: 16 mutations (24.24%), D: 8 mutations (12.12%), A: 3 mutations (4.54%) and D&E: 1 mutation (1.52%). No mutation was detected in probe C.

Conclusions: Mutations associated with RIF resistance in the rpoB gene of M. tuberculosis detected by the Xpert® MTB/RIF method were mainly found in probe E (codons 529-533). On the other hand, no mutation was detected in probe C.

Introducción. La tuberculosis continúa siendo un problema de salud pública agravado por la resistencia de Mycobacterium tuberculosis a los fármacos. Más del 95% de cepas de M. tuberculosis resistentes a rifampicina (RIF) poseen mutaciones en una región del gen rpoB. Xpert® MTB/RIF es un sistema de biología molecular que, mediante 5 sondas (A, B, C, D, E) que conforman las secuencias del gen rpoB, permite identificar mutaciones en la región determinante de resistencia a la RIF de este gen.

Objetivo. Describir la distribución y frecuencia de potenciales mutaciones asociadas con la resistencia a RIF en el gen rpoB de M. tuberculosis detectadas en muestras pulmonares y extrapulmonares usando el método Xpert® MTB/RIF.

Materiales y métodos. Estudio retrospectivo. Se analizaron 66 muestras positivas para M. tuberculosis resistente a RIF procesadas por el sistema GeneXpert MTB/RIF entre enero de 2011 y julio de 2019 en un hospital universitario de Medellín, Colombia. De acuerdo con el software Dx System del instrumento GenXpert, se determinó que había una potencial mutación y resistencia a RIF si las sondas no se unían a su secuencia complementaria natural o si se presentaba un atraso en la unión (delta C_T >4) en relación con las otras sondas por presencia de una secuencia anormal. Los datos se analizaron mediante estadística descriptiva.

Resultados. De las 66 muestras (48 pulmonares y 18 extrapulmonares), el 63.64% eran de hombres y la edad media de los participantes fue 39.60±17.69 años. La frecuencia y distribución de mutaciones fue la siguiente: sonda E: 38 mutaciones (57.58%), sonda B: 16 (24.24%), sonda D: 8 (12.12%), sonda A: 3 (4.54%) y la combinación de las sondas D&E: 1 (1.52%). No se detectó ninguna mutación en la sonda C.

Conclusiones. Las mutaciones asociadas a la resistencia a RIF en el gen rpoB de M. tuberculosis detectadas por el método Xpert® MTB/RIF se encontraron principalmente en la sonda E (codones 529-533). Por el contrario, en la sonda C no se no se detectó ninguna mutación.

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