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Viability PCR shows that non-ocular surfaces could contribute to transmission of Chlamydia trachomatis infection in trachoma

BACKGROUND: The presence of Chlamydia trachomatis (Ct) DNA at non-ocular sites suggests that these sites may represent plausible routes of Ct transmission in trachoma. However, qPCR cannot discriminate between DNA from viable and non-viable bacteria. Here we use a propodium monoazide based viability...

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Autores principales: Versteeg, Bart, Vasileva, Hristina, Houghton, Joanna, Last, Anna, Shafi Abdurahman, Oumer, Sarah, Virginia, Macleod, David, Solomon, Anthony W., Holland, Martin J., Thomson, Nicholas, Burton, Matthew J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7384675/
https://www.ncbi.nlm.nih.gov/pubmed/32667914
http://dx.doi.org/10.1371/journal.pntd.0008449
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author Versteeg, Bart
Vasileva, Hristina
Houghton, Joanna
Last, Anna
Shafi Abdurahman, Oumer
Sarah, Virginia
Macleod, David
Solomon, Anthony W.
Holland, Martin J.
Thomson, Nicholas
Burton, Matthew J.
author_facet Versteeg, Bart
Vasileva, Hristina
Houghton, Joanna
Last, Anna
Shafi Abdurahman, Oumer
Sarah, Virginia
Macleod, David
Solomon, Anthony W.
Holland, Martin J.
Thomson, Nicholas
Burton, Matthew J.
author_sort Versteeg, Bart
collection PubMed
description BACKGROUND: The presence of Chlamydia trachomatis (Ct) DNA at non-ocular sites suggests that these sites may represent plausible routes of Ct transmission in trachoma. However, qPCR cannot discriminate between DNA from viable and non-viable bacteria. Here we use a propodium monoazide based viability PCR to investigate how long Ct remains viable at non-ocular sites under laboratory-controlled conditions. METHODS: Cultured Ct stocks (strain A2497) were diluted to final concentrations of 1000, 100, 10 and 1 omcB copies/μL and applied to plastic, woven mat, cotton cloth and pig skin. Swabs were then systemically collected from each surface and tested for the presence Ct DNA using qPCR. If Ct DNA was recovered, Ct viability was assessed over time by spiking multiple areas of the same surface type with the same final concentrations. Swabs were collected from each surface at 0, 2, 4, 6, 8 and 24 hours after spiking. Viability PCR was used to determine Ct viability at each timepoint. RESULTS: We were able to detect Ct DNA on all surfaces except the woven mat. Total Ct DNA remained detectable and stable over 24 hours for all concentrations applied to plastic, pig skin and cotton cloth. The amount of viable Ct decreased over time. For plastic and skin surfaces, only those where concentrations of 100 or 1000 omcB copies/μL were applied still had viable loads detectable after 24 hours. Cotton cloth showed a more rapid decrease and only those where concentrations of 1000 omcB copies/μL were applied still had viable DNA detectable after 24 hours. CONCLUSION: Plastic, cotton cloth and skin may contribute to transmission of the Ct strains that cause trachoma, by acting as sites where reservoirs of bacteria are deposited and later collected and transferred mechanically into previously uninfected eyes.
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spelling pubmed-73846752020-08-05 Viability PCR shows that non-ocular surfaces could contribute to transmission of Chlamydia trachomatis infection in trachoma Versteeg, Bart Vasileva, Hristina Houghton, Joanna Last, Anna Shafi Abdurahman, Oumer Sarah, Virginia Macleod, David Solomon, Anthony W. Holland, Martin J. Thomson, Nicholas Burton, Matthew J. PLoS Negl Trop Dis Research Article BACKGROUND: The presence of Chlamydia trachomatis (Ct) DNA at non-ocular sites suggests that these sites may represent plausible routes of Ct transmission in trachoma. However, qPCR cannot discriminate between DNA from viable and non-viable bacteria. Here we use a propodium monoazide based viability PCR to investigate how long Ct remains viable at non-ocular sites under laboratory-controlled conditions. METHODS: Cultured Ct stocks (strain A2497) were diluted to final concentrations of 1000, 100, 10 and 1 omcB copies/μL and applied to plastic, woven mat, cotton cloth and pig skin. Swabs were then systemically collected from each surface and tested for the presence Ct DNA using qPCR. If Ct DNA was recovered, Ct viability was assessed over time by spiking multiple areas of the same surface type with the same final concentrations. Swabs were collected from each surface at 0, 2, 4, 6, 8 and 24 hours after spiking. Viability PCR was used to determine Ct viability at each timepoint. RESULTS: We were able to detect Ct DNA on all surfaces except the woven mat. Total Ct DNA remained detectable and stable over 24 hours for all concentrations applied to plastic, pig skin and cotton cloth. The amount of viable Ct decreased over time. For plastic and skin surfaces, only those where concentrations of 100 or 1000 omcB copies/μL were applied still had viable loads detectable after 24 hours. Cotton cloth showed a more rapid decrease and only those where concentrations of 1000 omcB copies/μL were applied still had viable DNA detectable after 24 hours. CONCLUSION: Plastic, cotton cloth and skin may contribute to transmission of the Ct strains that cause trachoma, by acting as sites where reservoirs of bacteria are deposited and later collected and transferred mechanically into previously uninfected eyes. Public Library of Science 2020-07-15 /pmc/articles/PMC7384675/ /pubmed/32667914 http://dx.doi.org/10.1371/journal.pntd.0008449 Text en © 2020 Versteeg 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 (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Versteeg, Bart
Vasileva, Hristina
Houghton, Joanna
Last, Anna
Shafi Abdurahman, Oumer
Sarah, Virginia
Macleod, David
Solomon, Anthony W.
Holland, Martin J.
Thomson, Nicholas
Burton, Matthew J.
Viability PCR shows that non-ocular surfaces could contribute to transmission of Chlamydia trachomatis infection in trachoma
title Viability PCR shows that non-ocular surfaces could contribute to transmission of Chlamydia trachomatis infection in trachoma
title_full Viability PCR shows that non-ocular surfaces could contribute to transmission of Chlamydia trachomatis infection in trachoma
title_fullStr Viability PCR shows that non-ocular surfaces could contribute to transmission of Chlamydia trachomatis infection in trachoma
title_full_unstemmed Viability PCR shows that non-ocular surfaces could contribute to transmission of Chlamydia trachomatis infection in trachoma
title_short Viability PCR shows that non-ocular surfaces could contribute to transmission of Chlamydia trachomatis infection in trachoma
title_sort viability pcr shows that non-ocular surfaces could contribute to transmission of chlamydia trachomatis infection in trachoma
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7384675/
https://www.ncbi.nlm.nih.gov/pubmed/32667914
http://dx.doi.org/10.1371/journal.pntd.0008449
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