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Real-Time PCR Validation for Mycobacterium tuberculosis Complex Detection Targeting IS6110 Directly From Bovine Lymph Nodes

Rapid and accurate diagnostic tools, such as Real-Time PCR (qPCR), need to be implemented as a confirmatory test in the framework of bovine tuberculosis (bTB) surveillance and control programs, shortening the turnaround time to confirm bTB infection. The present study aimed to evaluate a direct qPCR...

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Autores principales: Sánchez-Carvajal, José María, Galán-Relaño, Ángela, Ruedas-Torres, Inés, Jurado-Martos, Francisco, Larenas-Muñoz, Fernanda, Vera, Eduardo, Gómez-Gascón, Lidia, Cardoso-Toset, Fernando, Rodríguez-Gómez, Irene Magdalena, Maldonado, Alfonso, Carrasco, Librado, Tarradas, Carmen, Gómez-Laguna, Jaime, Luque, Inmaculada
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8109245/
https://www.ncbi.nlm.nih.gov/pubmed/33981742
http://dx.doi.org/10.3389/fvets.2021.643111
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author Sánchez-Carvajal, José María
Galán-Relaño, Ángela
Ruedas-Torres, Inés
Jurado-Martos, Francisco
Larenas-Muñoz, Fernanda
Vera, Eduardo
Gómez-Gascón, Lidia
Cardoso-Toset, Fernando
Rodríguez-Gómez, Irene Magdalena
Maldonado, Alfonso
Carrasco, Librado
Tarradas, Carmen
Gómez-Laguna, Jaime
Luque, Inmaculada
author_facet Sánchez-Carvajal, José María
Galán-Relaño, Ángela
Ruedas-Torres, Inés
Jurado-Martos, Francisco
Larenas-Muñoz, Fernanda
Vera, Eduardo
Gómez-Gascón, Lidia
Cardoso-Toset, Fernando
Rodríguez-Gómez, Irene Magdalena
Maldonado, Alfonso
Carrasco, Librado
Tarradas, Carmen
Gómez-Laguna, Jaime
Luque, Inmaculada
author_sort Sánchez-Carvajal, José María
collection PubMed
description Rapid and accurate diagnostic tools, such as Real-Time PCR (qPCR), need to be implemented as a confirmatory test in the framework of bovine tuberculosis (bTB) surveillance and control programs, shortening the turnaround time to confirm bTB infection. The present study aimed to evaluate a direct qPCR from fresh tissue samples targeting the insertion sequence IS6110 using individually homogenized bovine lymph nodes compared with microbiological culture. Retropharyngeal, tracheobronchial, and mesenteric lymph nodes fresh tissue samples (n = 687) were collected from 230 different cattle carcasses at the slaughterhouse. Only 23 of the 230 examined animals showed tuberculosis-like lesions, with 62 of 230 considered as positive. Among these 62 animals, 61 resulted as culture-positive, whereas 48 were qPCR-positive. Thus, this qPCR targeting IS6110 showed an apparent diagnostic sensitivity and specificity values of 77.1% [95% confidence interval (CI): 66.5–87.6%] and 99.4% (95% CI: 98.3–100.6%), respectively, and a positive predictive value of 97.9% (95% CI: 93.9–102.0%) and negative predictive value of 92.3% (95% CI: 88.4–96.2%). Positive and negative likelihood ratios were 130.2 and 0.2, respectively, and the agreement between microbiological culture and this qPCR was almost perfect (κ = 0.82). These results highlight this qPCR targeting IS6110 as a suitable complementary method to confirm bTB in animals with either tuberculosis-like lesions or non-tuberculosis-like lesions, decreasing the number of samples subjected to microbiological culture and, hence, its overall associated costs and the turnaround time (under 48 h) to confirm bTB infection. Besides, sampling mesenteric lymph node, which is uncommonly sampled, together with tracheobronchial and retropharyngeal ones, is advisable during postmortem inspection in bTB surveillance programs at the slaughterhouse, especially in areas with a low bTB prevalence scenario.
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spelling pubmed-81092452021-05-11 Real-Time PCR Validation for Mycobacterium tuberculosis Complex Detection Targeting IS6110 Directly From Bovine Lymph Nodes Sánchez-Carvajal, José María Galán-Relaño, Ángela Ruedas-Torres, Inés Jurado-Martos, Francisco Larenas-Muñoz, Fernanda Vera, Eduardo Gómez-Gascón, Lidia Cardoso-Toset, Fernando Rodríguez-Gómez, Irene Magdalena Maldonado, Alfonso Carrasco, Librado Tarradas, Carmen Gómez-Laguna, Jaime Luque, Inmaculada Front Vet Sci Veterinary Science Rapid and accurate diagnostic tools, such as Real-Time PCR (qPCR), need to be implemented as a confirmatory test in the framework of bovine tuberculosis (bTB) surveillance and control programs, shortening the turnaround time to confirm bTB infection. The present study aimed to evaluate a direct qPCR from fresh tissue samples targeting the insertion sequence IS6110 using individually homogenized bovine lymph nodes compared with microbiological culture. Retropharyngeal, tracheobronchial, and mesenteric lymph nodes fresh tissue samples (n = 687) were collected from 230 different cattle carcasses at the slaughterhouse. Only 23 of the 230 examined animals showed tuberculosis-like lesions, with 62 of 230 considered as positive. Among these 62 animals, 61 resulted as culture-positive, whereas 48 were qPCR-positive. Thus, this qPCR targeting IS6110 showed an apparent diagnostic sensitivity and specificity values of 77.1% [95% confidence interval (CI): 66.5–87.6%] and 99.4% (95% CI: 98.3–100.6%), respectively, and a positive predictive value of 97.9% (95% CI: 93.9–102.0%) and negative predictive value of 92.3% (95% CI: 88.4–96.2%). Positive and negative likelihood ratios were 130.2 and 0.2, respectively, and the agreement between microbiological culture and this qPCR was almost perfect (κ = 0.82). These results highlight this qPCR targeting IS6110 as a suitable complementary method to confirm bTB in animals with either tuberculosis-like lesions or non-tuberculosis-like lesions, decreasing the number of samples subjected to microbiological culture and, hence, its overall associated costs and the turnaround time (under 48 h) to confirm bTB infection. Besides, sampling mesenteric lymph node, which is uncommonly sampled, together with tracheobronchial and retropharyngeal ones, is advisable during postmortem inspection in bTB surveillance programs at the slaughterhouse, especially in areas with a low bTB prevalence scenario. Frontiers Media S.A. 2021-04-26 /pmc/articles/PMC8109245/ /pubmed/33981742 http://dx.doi.org/10.3389/fvets.2021.643111 Text en Copyright © 2021 Sánchez-Carvajal, Galán-Relaño, Ruedas-Torres, Jurado-Martos, Larenas-Muñoz, Vera, Gómez-Gascón, Cardoso-Toset, Rodríguez-Gómez, Maldonado, Carrasco, Tarradas, Gómez-Laguna and Luque. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Veterinary Science
Sánchez-Carvajal, José María
Galán-Relaño, Ángela
Ruedas-Torres, Inés
Jurado-Martos, Francisco
Larenas-Muñoz, Fernanda
Vera, Eduardo
Gómez-Gascón, Lidia
Cardoso-Toset, Fernando
Rodríguez-Gómez, Irene Magdalena
Maldonado, Alfonso
Carrasco, Librado
Tarradas, Carmen
Gómez-Laguna, Jaime
Luque, Inmaculada
Real-Time PCR Validation for Mycobacterium tuberculosis Complex Detection Targeting IS6110 Directly From Bovine Lymph Nodes
title Real-Time PCR Validation for Mycobacterium tuberculosis Complex Detection Targeting IS6110 Directly From Bovine Lymph Nodes
title_full Real-Time PCR Validation for Mycobacterium tuberculosis Complex Detection Targeting IS6110 Directly From Bovine Lymph Nodes
title_fullStr Real-Time PCR Validation for Mycobacterium tuberculosis Complex Detection Targeting IS6110 Directly From Bovine Lymph Nodes
title_full_unstemmed Real-Time PCR Validation for Mycobacterium tuberculosis Complex Detection Targeting IS6110 Directly From Bovine Lymph Nodes
title_short Real-Time PCR Validation for Mycobacterium tuberculosis Complex Detection Targeting IS6110 Directly From Bovine Lymph Nodes
title_sort real-time pcr validation for mycobacterium tuberculosis complex detection targeting is6110 directly from bovine lymph nodes
topic Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8109245/
https://www.ncbi.nlm.nih.gov/pubmed/33981742
http://dx.doi.org/10.3389/fvets.2021.643111
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