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Development of a Real-Time PCR for Identification of Brachyspira Species in Human Colonic Biopsies

BACKGROUND: Brachyspira species are fastidious anaerobic microorganisms, that infect the colon of various animals. The genus contains both important pathogens of livestock as well as commensals. Two species are known to infect humans: B. aalborgi and B. pilosicoli. There is some evidence suggesting...

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Autores principales: Westerman, Laurens J., Stel, Herbert V., Schipper, Marguerite E. I., Bakker, Leendert J., Neefjes-Borst, Eskelina A., van den Brande, Jan H. M., Boel, Edwin C. H., Seldenrijk, Kees A., Siersema, Peter D., Bonten, Marc J. M., Kusters, Johannes G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3527525/
https://www.ncbi.nlm.nih.gov/pubmed/23284968
http://dx.doi.org/10.1371/journal.pone.0052281
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author Westerman, Laurens J.
Stel, Herbert V.
Schipper, Marguerite E. I.
Bakker, Leendert J.
Neefjes-Borst, Eskelina A.
van den Brande, Jan H. M.
Boel, Edwin C. H.
Seldenrijk, Kees A.
Siersema, Peter D.
Bonten, Marc J. M.
Kusters, Johannes G.
author_facet Westerman, Laurens J.
Stel, Herbert V.
Schipper, Marguerite E. I.
Bakker, Leendert J.
Neefjes-Borst, Eskelina A.
van den Brande, Jan H. M.
Boel, Edwin C. H.
Seldenrijk, Kees A.
Siersema, Peter D.
Bonten, Marc J. M.
Kusters, Johannes G.
author_sort Westerman, Laurens J.
collection PubMed
description BACKGROUND: Brachyspira species are fastidious anaerobic microorganisms, that infect the colon of various animals. The genus contains both important pathogens of livestock as well as commensals. Two species are known to infect humans: B. aalborgi and B. pilosicoli. There is some evidence suggesting that the veterinary pathogenic B. pilosicoli is a potential zoonotic agent, however, since diagnosis in humans is based on histopathology of colon biopsies, species identification is not routinely performed in human materials. METHODS: The study population comprised 57 patients with microscopic evidence of Brachyspira infection and 26 patients with no histopathological evidence of Brachyspira infection. Concomitant faecal samples were available from three infected patients. Based on publically available 16S rDNA gene sequences of all Brachyspira species, species-specific primer sets were designed. DNA was extracted and tested by real-time PCR and 16S rDNA was sequenced. RESULTS: Sensitivity and specificity for identification of Brachyspira species in colon biopsies was 100% and 87.7% respectively. Sequencing revealed B. pilosicoli in 15.4% of patients, B. aalborgi in 76.9% and a third species, tentatively named “Brachyspira hominis”, in 26.2%. Ten patients (12.3%) had a double and two (3.1%) a triple infection. The presence of Brachyspira pilosicoli was significantly associated with inflammatory changes in the colon-biopsy (p = 0.028). CONCLUSIONS: This newly designed PCR allows for sub-differentiation of Brachyspira species in patient material and thus allows large-scaled surveillance studies to elucidate the pathogenicity of human Brachyspira infections. One-third of affected patients appeared to be infected with a novel species.
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spelling pubmed-35275252013-01-02 Development of a Real-Time PCR for Identification of Brachyspira Species in Human Colonic Biopsies Westerman, Laurens J. Stel, Herbert V. Schipper, Marguerite E. I. Bakker, Leendert J. Neefjes-Borst, Eskelina A. van den Brande, Jan H. M. Boel, Edwin C. H. Seldenrijk, Kees A. Siersema, Peter D. Bonten, Marc J. M. Kusters, Johannes G. PLoS One Research Article BACKGROUND: Brachyspira species are fastidious anaerobic microorganisms, that infect the colon of various animals. The genus contains both important pathogens of livestock as well as commensals. Two species are known to infect humans: B. aalborgi and B. pilosicoli. There is some evidence suggesting that the veterinary pathogenic B. pilosicoli is a potential zoonotic agent, however, since diagnosis in humans is based on histopathology of colon biopsies, species identification is not routinely performed in human materials. METHODS: The study population comprised 57 patients with microscopic evidence of Brachyspira infection and 26 patients with no histopathological evidence of Brachyspira infection. Concomitant faecal samples were available from three infected patients. Based on publically available 16S rDNA gene sequences of all Brachyspira species, species-specific primer sets were designed. DNA was extracted and tested by real-time PCR and 16S rDNA was sequenced. RESULTS: Sensitivity and specificity for identification of Brachyspira species in colon biopsies was 100% and 87.7% respectively. Sequencing revealed B. pilosicoli in 15.4% of patients, B. aalborgi in 76.9% and a third species, tentatively named “Brachyspira hominis”, in 26.2%. Ten patients (12.3%) had a double and two (3.1%) a triple infection. The presence of Brachyspira pilosicoli was significantly associated with inflammatory changes in the colon-biopsy (p = 0.028). CONCLUSIONS: This newly designed PCR allows for sub-differentiation of Brachyspira species in patient material and thus allows large-scaled surveillance studies to elucidate the pathogenicity of human Brachyspira infections. One-third of affected patients appeared to be infected with a novel species. Public Library of Science 2012-12-20 /pmc/articles/PMC3527525/ /pubmed/23284968 http://dx.doi.org/10.1371/journal.pone.0052281 Text en © 2012 Westerman et al http://creativecommons.org/licenses/by/4.0/ 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 properly credited.
spellingShingle Research Article
Westerman, Laurens J.
Stel, Herbert V.
Schipper, Marguerite E. I.
Bakker, Leendert J.
Neefjes-Borst, Eskelina A.
van den Brande, Jan H. M.
Boel, Edwin C. H.
Seldenrijk, Kees A.
Siersema, Peter D.
Bonten, Marc J. M.
Kusters, Johannes G.
Development of a Real-Time PCR for Identification of Brachyspira Species in Human Colonic Biopsies
title Development of a Real-Time PCR for Identification of Brachyspira Species in Human Colonic Biopsies
title_full Development of a Real-Time PCR for Identification of Brachyspira Species in Human Colonic Biopsies
title_fullStr Development of a Real-Time PCR for Identification of Brachyspira Species in Human Colonic Biopsies
title_full_unstemmed Development of a Real-Time PCR for Identification of Brachyspira Species in Human Colonic Biopsies
title_short Development of a Real-Time PCR for Identification of Brachyspira Species in Human Colonic Biopsies
title_sort development of a real-time pcr for identification of brachyspira species in human colonic biopsies
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3527525/
https://www.ncbi.nlm.nih.gov/pubmed/23284968
http://dx.doi.org/10.1371/journal.pone.0052281
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