CagA gene EPIYA motif genetic characterization from Colombian Helicobacter pylori isolates: Standardization of a molecular test for rapid clinical laboratory detection

Eliana Rocio Rodriguez Gómez; William Otero Regino; Pedro A. Monterrey; Alba Alicia Trespalacios Rangel

CagA gene EPIYA motif genetic characterization from Colombian Helicobacter pylori isolates: Standardization of a molecular test for rapid clinical laboratory detection

Título traducido de la contribución: Caracterización genética del motivo EPIYA del gen cagA en aislamientos colombianos de Helicobacter pylori: Estandarización de una prueba molecular para la detección rápida en laboratorio clínico

Eliana Rocio Rodriguez Gómez, William Otero Regino, Pedro A. Monterrey, Alba Alicia Trespalacios Rangel

Producción: Contribución a una revista › Artículo › revisión exhaustiva

13 Citas (Scopus)

Resumen

El objetivo de este trabajo fue determinar los motivos EPIYA del gen cagA presentes en aislamientos colombianos de Helicobacter pylori mediante una prueba molecular rápida y confiable. Se analizó el ADN de ochenta y cinco cepas de Helicobacter pylori-cagA positivas. Las cepas se obtuvieron de pacientes diagnosticados con dispepsia funcional en la Clínica Fundadores en Bogotá. La región 3' del gen cagA se amplificó mediante reacción en cadena de la polimerasa (PCR) convencional. Los amplicones obtenidos se secuenciaron mediante el método de Sanger y se analizaron con herramientas bioinformáticas. Además, se determinó un coeficiente de correlación de Spearman significativo entre la edad de los pacientes y el número de repeticiones de EPIYA-C; con valores p < 0,05 considerados significativos. Las estimaciones se obtuvieron utilizando un IC del 95%. Se amplificó la región variable 3' del gen cagA y los productos de PCR de los siguientes tamaños correspondieron a los siguientes motivos EPIYA: 400 pb: EPIYA AB, 500 pb: EPIYA ABC, 600 pb: EPIYA ABCC y 700 pb: ABCCC. Se observó una sola banda de PCR para 58 de 85 aislados de Helicobacter pylori , con un motivo de distribución EPIYA de la siguiente manera: 7/85 AB (8,2%), 34/85 ABC (40%), 26/85 ABCC (30,6%) y 18/85 ABCCC (21,2%). Sin embargo, en 27 de 85 aislados de Helicobacter pylori , se observaron dos o más bandas, donde el genotipo cagA más predominante fue ABC-ABCC (26%, 7/27) y ABCC-ABCCC (22,2%, 6/27). Se observó una proporcionalidad directa entre el número de repeticiones de EPIYA-C y el aumento de la edad de los pacientes, encontrándose un mayor número de repeticiones de EPIYA ABCC y ABCCC en la población mayor de 50 años. Todos los aislados fueron del tipo Western cagA y el 51,8% presentó múltiples repeticiones de EPIYA-C. Esta prueba molecular estandarizada permitió identificar el número de motivos EPIYA C según el tamaño de la banda.

Título traducido de la contribución	Caracterización genética del motivo EPIYA del gen cagA en aislamientos colombianos de Helicobacter pylori: Estandarización de una prueba molecular para la detección rápida en laboratorio clínico
Idioma original	Inglés
Número de artículo	e0227275
Páginas (desde-hasta)	1-12
Número de páginas	12
Publicación	PLoS ONE
Volumen	15
N.º	1
Estado	Publicada - 01 ene. 2020

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title = "CagA gene EPIYA motif genetic characterization from Colombian Helicobacter pylori isolates: Standardization of a molecular test for rapid clinical laboratory detection",

abstract = "The aim of this work was to determine current cagA gene EPIYA motifs present in Colombian Helicobacter pylori isolates using a fast and reliable molecular test. DNA from eightyfive Helicobacter pylori-cagA positive strains were analyzed. Strains were obtained from patients diagnosed with functional dyspepsia at Cl? nica Fundadores in Bogota . The 3' region of the cagA gene was amplified through conventional Polymerase Chain Reaction (PCR). Obtained amplicons were sequenced using the Sanger method and analyzed with bioinformatics tools. Additionally, a significant Spearman correlation coefficient was determined between the patients' age and the number of EPIYA-C repeats; with p values < 0.05 considered significant. Estimates were obtained using a 95\% CI. The 3 variable region of the cagA gene was amplified and PCR products of the following sizes corresponded to the following EPIYA motifs: 400 bp: EPIYA AB, 500 bp: EPIYA ABC, 600 bp: EPIYA ABCC and 700 bp: ABCCC. A single PCR band was observed for 58 out of 85 Helicobacter pylori isolates, with an EPIYA distribution motif as follows: 7/85 AB (8.2\%), 34/85 ABC (40\%), 26/85 ABCC (30.6\%) and 18/85 ABCCC (21.2\%). However, in 27 out of 85 Helicobacter pylori isolates, two or more bands were observed, where the most predominant cagA genotype were ABC-ABCC (26\%, 7/27) and ABCC-ABCCC (22.2\%, 6/27). A direct proportionality between the number of EPIYA-C repeats and an increase in the patients' age was observed, finding a greater number of EPIYA ABCC and ABCCC repeats in the population over 50 years old. All isolates were of the Western cagA type and 51.8\% of them were found to have multiple EPIYA-C repeats. These standardized molecular test allowed to identify the number of EPIYA C motifs based on band size.",

author = "G{\'o}mez, \{Eliana Rocio Rodriguez\} and Regino, \{William Otero\} and Monterrey, \{Pedro A.\} and Rangel, \{Alba Alicia Trespalacios\}",

note = "Publisher Copyright: {\textcopyright} 2020 Rodriguez Gomez et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",

year = "2020",

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CagA gene EPIYA motif genetic characterization from Colombian Helicobacter pylori isolates: Standardization of a molecular test for rapid clinical laboratory detection. / Gómez, Eliana Rocio Rodriguez; Regino, William Otero; Monterrey, Pedro A. et al.
En: PLoS ONE, Vol. 15, N.º 1, e0227275, 01.01.2020, p. 1-12.

Producción: Contribución a una revista › Artículo › revisión exhaustiva

TY - JOUR

T1 - CagA gene EPIYA motif genetic characterization from Colombian Helicobacter pylori isolates

T2 - Standardization of a molecular test for rapid clinical laboratory detection

AU - Gómez, Eliana Rocio Rodriguez

AU - Regino, William Otero

AU - Monterrey, Pedro A.

AU - Rangel, Alba Alicia Trespalacios

N1 - Publisher Copyright: © 2020 Rodriguez Gomez et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

PY - 2020/1/1

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N2 - The aim of this work was to determine current cagA gene EPIYA motifs present in Colombian Helicobacter pylori isolates using a fast and reliable molecular test. DNA from eightyfive Helicobacter pylori-cagA positive strains were analyzed. Strains were obtained from patients diagnosed with functional dyspepsia at Cl? nica Fundadores in Bogota . The 3' region of the cagA gene was amplified through conventional Polymerase Chain Reaction (PCR). Obtained amplicons were sequenced using the Sanger method and analyzed with bioinformatics tools. Additionally, a significant Spearman correlation coefficient was determined between the patients' age and the number of EPIYA-C repeats; with p values < 0.05 considered significant. Estimates were obtained using a 95% CI. The 3 variable region of the cagA gene was amplified and PCR products of the following sizes corresponded to the following EPIYA motifs: 400 bp: EPIYA AB, 500 bp: EPIYA ABC, 600 bp: EPIYA ABCC and 700 bp: ABCCC. A single PCR band was observed for 58 out of 85 Helicobacter pylori isolates, with an EPIYA distribution motif as follows: 7/85 AB (8.2%), 34/85 ABC (40%), 26/85 ABCC (30.6%) and 18/85 ABCCC (21.2%). However, in 27 out of 85 Helicobacter pylori isolates, two or more bands were observed, where the most predominant cagA genotype were ABC-ABCC (26%, 7/27) and ABCC-ABCCC (22.2%, 6/27). A direct proportionality between the number of EPIYA-C repeats and an increase in the patients' age was observed, finding a greater number of EPIYA ABCC and ABCCC repeats in the population over 50 years old. All isolates were of the Western cagA type and 51.8% of them were found to have multiple EPIYA-C repeats. These standardized molecular test allowed to identify the number of EPIYA C motifs based on band size.

AB - The aim of this work was to determine current cagA gene EPIYA motifs present in Colombian Helicobacter pylori isolates using a fast and reliable molecular test. DNA from eightyfive Helicobacter pylori-cagA positive strains were analyzed. Strains were obtained from patients diagnosed with functional dyspepsia at Cl? nica Fundadores in Bogota . The 3' region of the cagA gene was amplified through conventional Polymerase Chain Reaction (PCR). Obtained amplicons were sequenced using the Sanger method and analyzed with bioinformatics tools. Additionally, a significant Spearman correlation coefficient was determined between the patients' age and the number of EPIYA-C repeats; with p values < 0.05 considered significant. Estimates were obtained using a 95% CI. The 3 variable region of the cagA gene was amplified and PCR products of the following sizes corresponded to the following EPIYA motifs: 400 bp: EPIYA AB, 500 bp: EPIYA ABC, 600 bp: EPIYA ABCC and 700 bp: ABCCC. A single PCR band was observed for 58 out of 85 Helicobacter pylori isolates, with an EPIYA distribution motif as follows: 7/85 AB (8.2%), 34/85 ABC (40%), 26/85 ABCC (30.6%) and 18/85 ABCCC (21.2%). However, in 27 out of 85 Helicobacter pylori isolates, two or more bands were observed, where the most predominant cagA genotype were ABC-ABCC (26%, 7/27) and ABCC-ABCCC (22.2%, 6/27). A direct proportionality between the number of EPIYA-C repeats and an increase in the patients' age was observed, finding a greater number of EPIYA ABCC and ABCCC repeats in the population over 50 years old. All isolates were of the Western cagA type and 51.8% of them were found to have multiple EPIYA-C repeats. These standardized molecular test allowed to identify the number of EPIYA C motifs based on band size.

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