Uso de Chelex-100 para o diagnóstico molecular de cinco patógenos animais

Molecular tools have improved conventional veterinary diagnosis. Acid nucleic extraction is a key step for downstream applications. This work aimed to compare the DNA extraction method Chelex-100 resin (M1) with Whatman® cards (M2), phenol-chloroform (M3), or commercial kits (M4), and to determine t...

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Autores principales: TOMAZIC, Mariela Luján, HAMER, Micaela, BUSTOS, Carla Paola, ARREGUI, Matías, ASCENCIO, Mariano, SARAULLO, Vanina, WATANABE, Olivia, BRIHUEGA, Bibiana, RODRIGUEZ, Anabel Elisa, GRUNE-LOFFLER, Sylvia
Formato: Artículo revista
Lenguaje:Inglés
Español
Publicado: Universidad Nacional del Litoral 2021
Materias:
Veterinary
Diagnosis
DNA
Chelex-100
PCR
Veterinaria
Diagnóstico
ADN
Acceso en línea:https://bibliotecavirtual.unl.edu.ar/publicaciones/index.php/FAVEveterinaria/article/view/9724
Aporte de:
Biblioteca Virtual - Publicaciones (UNL) de Universidad Nacional del Litoral
id I26-R133-article-9724
record_format ojs
institution Universidad Nacional del Litoral
institution_str I-26
repository_str R-133
container_title_str Biblioteca Virtual - Publicaciones (UNL)
language Inglés
Español
format Artículo revista
topic Veterinary
Diagnosis
DNA
Chelex-100
PCR
Veterinaria
Diagnóstico
Veterinaria
Diagnóstico
ADN
Chelex-100
PCR
spellingShingle Veterinary
Diagnosis
DNA
Chelex-100
PCR
Veterinaria
Diagnóstico
Veterinaria
Diagnóstico
ADN
Chelex-100
PCR
TOMAZIC, Mariela Luján
HAMER, Micaela
BUSTOS, Carla Paola
ARREGUI, Matías
ASCENCIO, Mariano
SARAULLO, Vanina
WATANABE, Olivia
BRIHUEGA, Bibiana
RODRIGUEZ, Anabel Elisa
GRUNE-LOFFLER, Sylvia
Uso de Chelex-100 para o diagnóstico molecular de cinco patógenos animais
topic_facet Veterinary
Diagnosis
DNA
Chelex-100
PCR
Veterinaria
Diagnóstico
Veterinaria
Diagnóstico
ADN
Chelex-100
PCR
author TOMAZIC, Mariela Luján
HAMER, Micaela
BUSTOS, Carla Paola
ARREGUI, Matías
ASCENCIO, Mariano
SARAULLO, Vanina
WATANABE, Olivia
BRIHUEGA, Bibiana
RODRIGUEZ, Anabel Elisa
GRUNE-LOFFLER, Sylvia
author_facet TOMAZIC, Mariela Luján
HAMER, Micaela
BUSTOS, Carla Paola
ARREGUI, Matías
ASCENCIO, Mariano
SARAULLO, Vanina
WATANABE, Olivia
BRIHUEGA, Bibiana
RODRIGUEZ, Anabel Elisa
GRUNE-LOFFLER, Sylvia
author_sort TOMAZIC, Mariela Luján
title Uso de Chelex-100 para o diagnóstico molecular de cinco patógenos animais
title_short Uso de Chelex-100 para o diagnóstico molecular de cinco patógenos animais
title_full Uso de Chelex-100 para o diagnóstico molecular de cinco patógenos animais
title_fullStr Uso de Chelex-100 para o diagnóstico molecular de cinco patógenos animais
title_full_unstemmed Uso de Chelex-100 para o diagnóstico molecular de cinco patógenos animais
title_sort uso de chelex-100 para o diagnóstico molecular de cinco patógenos animais
description Molecular tools have improved conventional veterinary diagnosis. Acid nucleic extraction is a key step for downstream applications. This work aimed to compare the DNA extraction method Chelex-100 resin (M1) with Whatman® cards (M2), phenol-chloroform (M3), or commercial kits (M4), and to determine the most sensitive and inexpensive one for its diagnosis of animal pathogens that, despite their economic or zoonotic relevance, receive little attention. DNA was isolated from urine, organs, semen, blood and intestinal mucous, from the bacteria Leptospira interrogans serovar Pomona Pomona (by M1 and M2), Brucella melitensis (by M1, M3 and M4), and Salmonella ser. Abortusequi (by M1 and M4), and the parasites Leishmania spp. (by M1, M3 and M4), and Eimeria spp. (by M1 and M3), respectively. The sensitivity of each method was assayed by Polymerase Chain Reaction (PCR).  The M1 showed similar sensitivity for Salmonella ser. Abortusequi, Leishmania spp., and Eimeria spp., being better for L. interrogans serovar Pomona Pomona and slightly lower for B. melitensis. For the first time, a simple and economic method was successfully employed for extracting DNA from these animal pathogens, especially important in low-resource settings, contributing to the diagnosis of leptospirosis, brucellosis, leishmaniasis, and coccidiosis; as well as to the molecular epidemiology of salmonellosis in stallion from semen samples.
publisher Universidad Nacional del Litoral
publishDate 2021
url https://bibliotecavirtual.unl.edu.ar/publicaciones/index.php/FAVEveterinaria/article/view/9724
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first_indexed 2023-07-05T23:07:05Z
last_indexed 2023-07-05T23:07:05Z
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spelling I26-R133-article-97242021-04-12T16:14:18Z Uso de Chelex-100 para o diagnóstico molecular de cinco patógenos animais Uso de Chelex-100 para el diagnóstico molecular de cinco patógenos de animales Use of Chelex-100 for the molecular diagnosis of five animal pathogens TOMAZIC, Mariela Luján HAMER, Micaela BUSTOS, Carla Paola ARREGUI, Matías ASCENCIO, Mariano SARAULLO, Vanina WATANABE, Olivia BRIHUEGA, Bibiana RODRIGUEZ, Anabel Elisa GRUNE-LOFFLER, Sylvia Veterinary Diagnosis DNA Chelex-100 PCR Veterinaria Diagnóstico Veterinaria Diagnóstico ADN Chelex-100 PCR Molecular tools have improved conventional veterinary diagnosis. Acid nucleic extraction is a key step for downstream applications. This work aimed to compare the DNA extraction method Chelex-100 resin (M1) with Whatman® cards (M2), phenol-chloroform (M3), or commercial kits (M4), and to determine the most sensitive and inexpensive one for its diagnosis of animal pathogens that, despite their economic or zoonotic relevance, receive little attention. DNA was isolated from urine, organs, semen, blood and intestinal mucous, from the bacteria Leptospira interrogans serovar Pomona Pomona (by M1 and M2), Brucella melitensis (by M1, M3 and M4), and Salmonella ser. Abortusequi (by M1 and M4), and the parasites Leishmania spp. (by M1, M3 and M4), and Eimeria spp. (by M1 and M3), respectively. The sensitivity of each method was assayed by Polymerase Chain Reaction (PCR).  The M1 showed similar sensitivity for Salmonella ser. Abortusequi, Leishmania spp., and Eimeria spp., being better for L. interrogans serovar Pomona Pomona and slightly lower for B. melitensis. For the first time, a simple and economic method was successfully employed for extracting DNA from these animal pathogens, especially important in low-resource settings, contributing to the diagnosis of leptospirosis, brucellosis, leishmaniasis, and coccidiosis; as well as to the molecular epidemiology of salmonellosis in stallion from semen samples. Las técnicas moleculares han contribuido a mejorar el diagnóstico veterinario tradicional y la extracción de ácidos nucleicos es determinante. El objetivo de este trabajo fue comparar el método de extracción de ADN Chelex-100 (M1) con papel Whatman (M2), fenol-cloroformo (M3) o kits comerciales (M4), y determinar un método sensible y de bajo costo para el diagnóstico de patógenos de animales económica o zoonóticamente relevantes y que reciben poca atención. A partir de orina, órganos, semen, sangre y mucosa intestinal se extrajo el ADN de las bacterias Leptospira interrogans serovar Pomona Pomona (con M1 y M2), Brucella melitensis (con M1, M3 y M4), Salmonella ser. Abortusequi (M1 y M4), y de los parásitos Leishmania spp. (M1, M3 y M4) y Eimeria spp. (M1 y M3), respectivamente. La sensibilidad de los protocolos fue analizada por PCR. El método M1 demostró una sensibilidad similar para S. Abortusequi, Leishmania spp. y Eimeria spp., siendo mejor para L. interrogans y levemente menor para B. melitensis. Por primera vez se usó exitosamente en estos patógenos veterinarios un método simple y económico para extraer ADN, especialmente importante en laboratorios de bajos recursos económicos, contribuyendo al diagnóstico de leptospirosis, brucelosis, leishmaniasis y coccidiosis, así como también a la epidemiología molecular de salmonelosis en muestras de semen de caballos. As técnicas moleculares têm contribuído para melhorar o diagnóstico veterinário tradicional e a extração de ácido nucléico é crucial. O objetivo deste trabalho foi comparar o método de extração de DNA Chelex-100 (M1) com papel Whatman (M2), fenol-clorofórmio (M3) ou kits comerciais (M4), e determinar um método sensível e de baixo custo para o diagnóstico de patógenos animais economicamente ou zoonoticamente relevantes recebendo pouca atenção. Da urina, órgãos, sêmen, sangue e mucosa intestinal, foi extraído DNA das bactérias Leptospira interrogans sorovar Pomona Pomona (com M1 e M2), Brucella melitensis (com M1, M3 e M4), Salmonella ser. Abortusequi (M1 e M4), e da Leishmania spp. (M1, M3 e M4) e Eimeria spp. (M1 e M3), respectivamente. A sensibilidade dos protocolos foi analisada por PCR. O método M1 demonstrou sensibilidade semelhante para S. Abortusequi, Leishmania spp. e Eimeria spp., sendo melhor para L. interrogans e ligeiramente inferior para B. melitensis. Pela primeira vez, um método simples e barato de extração de DNA foi utilizado com sucesso nesses patógenos veterinários, especialmente importante em laboratórios com poucos recursos econômicos, contribuindo para o diagnóstico de leptospirose, brucelose, leishmaniose e coccidiose, bem como para a epidemiologia molecular de salmonelose em amostras de sêmen equino. Universidad Nacional del Litoral 2021-04-06 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Artículo original Original article application/pdf application/pdf https://bibliotecavirtual.unl.edu.ar/publicaciones/index.php/FAVEveterinaria/article/view/9724 10.14409/favecv.v20i1.9724 FAVE Sección Ciencias Veterinarias; Vol. 20 Núm. 1 (2021): FAVE Sección Ciencias Veterinarias; 26-33 FAVE Sección Ciencias Veterinarias; Vol. 20 No. 1 (2021): FAVE Sección Ciencias Veterinarias; 26-33 FAVE Sección Ciencias Veterinarias; v. 20 n. 1 (2021): FAVE Sección Ciencias Veterinarias; 26-33 2362-5589 1666-938X 10.14409/favecv.v20i1 eng spa https://bibliotecavirtual.unl.edu.ar/publicaciones/index.php/FAVEveterinaria/article/view/9724/13541 https://bibliotecavirtual.unl.edu.ar/publicaciones/index.php/FAVEveterinaria/article/view/9724/13542 Derechos de autor 2021 Mariela Luján TOMAZIC, Micaela HAMER, Carla Paola BUSTOS, Matías ARREGUI, Mariano ASCENCIO, Vanina SARAULLO, Olivia WATANABE, Bibiana BRIHUEGA, Anabel Elisa RODRIGUEZ, Sylvia GRUNE-LOFFLER

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