Cargando…
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...
Autores principales: | , , , , , , , , , , , , , |
---|---|
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 |
_version_ | 1783690216422244352 |
---|---|
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. |
format | Online Article Text |
id | pubmed-8109245 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT sanchezcarvajaljosemaria realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes AT galanrelanoangela realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes AT ruedastorresines realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes AT juradomartosfrancisco realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes AT larenasmunozfernanda realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes AT veraeduardo realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes AT gomezgasconlidia realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes AT cardosotosetfernando realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes AT rodriguezgomezirenemagdalena realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes AT maldonadoalfonso realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes AT carrascolibrado realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes AT tarradascarmen realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes AT gomezlagunajaime realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes AT luqueinmaculada realtimepcrvalidationformycobacteriumtuberculosiscomplexdetectiontargetingis6110directlyfrombovinelymphnodes |