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Quantification of Mycobacterium avium subspecies in pig tissues by real-time quantitative PCR

BACKGROUND: Mycobacterioses in animals cause economical losses and certain Mycobacterium avium subspecies are regarded as potential zoonotic agents. The evaluation of the zoonotic risk caused by M. avium subspecies requires information about the quantities of Mycobacterium strains in infected animal...

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Autores principales: Tirkkonen, Taneli, Nieminen, Timo, Ali-Vehmas, Terhi, Peltoniemi, Olli AT, Wellenberg, Gerard J, Pakarinen, Jaakko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3616872/
https://www.ncbi.nlm.nih.gov/pubmed/23518149
http://dx.doi.org/10.1186/1751-0147-55-26
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author Tirkkonen, Taneli
Nieminen, Timo
Ali-Vehmas, Terhi
Peltoniemi, Olli AT
Wellenberg, Gerard J
Pakarinen, Jaakko
author_facet Tirkkonen, Taneli
Nieminen, Timo
Ali-Vehmas, Terhi
Peltoniemi, Olli AT
Wellenberg, Gerard J
Pakarinen, Jaakko
author_sort Tirkkonen, Taneli
collection PubMed
description BACKGROUND: Mycobacterioses in animals cause economical losses and certain Mycobacterium avium subspecies are regarded as potential zoonotic agents. The evaluation of the zoonotic risk caused by M. avium subspecies requires information about the quantities of Mycobacterium strains in infected animals. Because M. avium subspecies in pig tissues are difficult or even impossible to quantify by culturing, we tested the suitability of a culture-independent real-time quantitative PCR (qPCR) assay for this purpose. METHODS: Mycobacterial DNA was extracted from porcine tissues by a novel method and quantified by Mycobacterium genus specific qPCR assay targeting the 16S rRNA gene. RESULTS: The response of the qPCR assay to the amount of M. avium subspecies avium mixed with porcine liver was linear in the range of approximately log10(5) to log10(7)Mycobacterium cells per 1 g of liver. The assay was validated with three other M. avium subspecies strains. When the assay was applied to porcine lymph nodes with or without visible lesions related to Mycobacterium avium subspecies infections, around 10(4)–10(7) mycobacterial genomes per gram of lymph nodes were detected. CONCLUSIONS: The qPCR assay was found to be suitable for the quantification of Mycobacterium avium subspecies in porcine lymph nodes and liver.
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spelling pubmed-36168722013-04-05 Quantification of Mycobacterium avium subspecies in pig tissues by real-time quantitative PCR Tirkkonen, Taneli Nieminen, Timo Ali-Vehmas, Terhi Peltoniemi, Olli AT Wellenberg, Gerard J Pakarinen, Jaakko Acta Vet Scand Research BACKGROUND: Mycobacterioses in animals cause economical losses and certain Mycobacterium avium subspecies are regarded as potential zoonotic agents. The evaluation of the zoonotic risk caused by M. avium subspecies requires information about the quantities of Mycobacterium strains in infected animals. Because M. avium subspecies in pig tissues are difficult or even impossible to quantify by culturing, we tested the suitability of a culture-independent real-time quantitative PCR (qPCR) assay for this purpose. METHODS: Mycobacterial DNA was extracted from porcine tissues by a novel method and quantified by Mycobacterium genus specific qPCR assay targeting the 16S rRNA gene. RESULTS: The response of the qPCR assay to the amount of M. avium subspecies avium mixed with porcine liver was linear in the range of approximately log10(5) to log10(7)Mycobacterium cells per 1 g of liver. The assay was validated with three other M. avium subspecies strains. When the assay was applied to porcine lymph nodes with or without visible lesions related to Mycobacterium avium subspecies infections, around 10(4)–10(7) mycobacterial genomes per gram of lymph nodes were detected. CONCLUSIONS: The qPCR assay was found to be suitable for the quantification of Mycobacterium avium subspecies in porcine lymph nodes and liver. BioMed Central 2013-03-22 /pmc/articles/PMC3616872/ /pubmed/23518149 http://dx.doi.org/10.1186/1751-0147-55-26 Text en Copyright © 2013 Tirkkonen et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Tirkkonen, Taneli
Nieminen, Timo
Ali-Vehmas, Terhi
Peltoniemi, Olli AT
Wellenberg, Gerard J
Pakarinen, Jaakko
Quantification of Mycobacterium avium subspecies in pig tissues by real-time quantitative PCR
title Quantification of Mycobacterium avium subspecies in pig tissues by real-time quantitative PCR
title_full Quantification of Mycobacterium avium subspecies in pig tissues by real-time quantitative PCR
title_fullStr Quantification of Mycobacterium avium subspecies in pig tissues by real-time quantitative PCR
title_full_unstemmed Quantification of Mycobacterium avium subspecies in pig tissues by real-time quantitative PCR
title_short Quantification of Mycobacterium avium subspecies in pig tissues by real-time quantitative PCR
title_sort quantification of mycobacterium avium subspecies in pig tissues by real-time quantitative pcr
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3616872/
https://www.ncbi.nlm.nih.gov/pubmed/23518149
http://dx.doi.org/10.1186/1751-0147-55-26
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