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Dried Saliva Spots: A Robust Method for Detecting Streptococcus pneumoniae Carriage by PCR

The earliest studies in the late 19th century on Streptococcus pneumoniae (S. pneumoniae) carriage used saliva as the primary specimen. However, interest in saliva declined after the sensitive mouse inoculation method was replaced by conventional culture, which made isolation of pneumococci from the...

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Autores principales: Krone, Cassandra L., Oja, Anna E., van de Groep, Kirsten, Sanders, Elisabeth A. M., Bogaert, Debby, Trzciński, Krzysztof
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4813204/
https://www.ncbi.nlm.nih.gov/pubmed/26959014
http://dx.doi.org/10.3390/ijms17030343
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author Krone, Cassandra L.
Oja, Anna E.
van de Groep, Kirsten
Sanders, Elisabeth A. M.
Bogaert, Debby
Trzciński, Krzysztof
author_facet Krone, Cassandra L.
Oja, Anna E.
van de Groep, Kirsten
Sanders, Elisabeth A. M.
Bogaert, Debby
Trzciński, Krzysztof
author_sort Krone, Cassandra L.
collection PubMed
description The earliest studies in the late 19th century on Streptococcus pneumoniae (S. pneumoniae) carriage used saliva as the primary specimen. However, interest in saliva declined after the sensitive mouse inoculation method was replaced by conventional culture, which made isolation of pneumococci from the highly polymicrobial oral cavity virtually impossible. Here, we tested the feasibility of using dried saliva spots (DSS) for studies on pneumococcal carriage. Saliva samples from children and pneumococcus-spiked saliva samples from healthy adults were applied to paper, dried, and stored, with and without desiccant, at temperatures ranging from −20 to 37 °C for up to 35 days. DNA extracted from DSS was tested with quantitative-PCR (qPCR) specifically for S. pneumoniae. When processed immediately after drying, the quantity of pneumococcal DNA detected in spiked DSS from adults matched the levels in freshly spiked raw saliva. Furthermore, pneumococcal DNA was stable in DSS stored with desiccant for up to one month over a broad range of temperatures. There were no differences in the results when spiking saliva with varied pneumococcal strains. The collection of saliva can be a particularly useful in surveillance studies conducted in remote settings, as it does not require trained personnel, and DSS are resilient to various transportation conditions.
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spelling pubmed-48132042016-04-06 Dried Saliva Spots: A Robust Method for Detecting Streptococcus pneumoniae Carriage by PCR Krone, Cassandra L. Oja, Anna E. van de Groep, Kirsten Sanders, Elisabeth A. M. Bogaert, Debby Trzciński, Krzysztof Int J Mol Sci Article The earliest studies in the late 19th century on Streptococcus pneumoniae (S. pneumoniae) carriage used saliva as the primary specimen. However, interest in saliva declined after the sensitive mouse inoculation method was replaced by conventional culture, which made isolation of pneumococci from the highly polymicrobial oral cavity virtually impossible. Here, we tested the feasibility of using dried saliva spots (DSS) for studies on pneumococcal carriage. Saliva samples from children and pneumococcus-spiked saliva samples from healthy adults were applied to paper, dried, and stored, with and without desiccant, at temperatures ranging from −20 to 37 °C for up to 35 days. DNA extracted from DSS was tested with quantitative-PCR (qPCR) specifically for S. pneumoniae. When processed immediately after drying, the quantity of pneumococcal DNA detected in spiked DSS from adults matched the levels in freshly spiked raw saliva. Furthermore, pneumococcal DNA was stable in DSS stored with desiccant for up to one month over a broad range of temperatures. There were no differences in the results when spiking saliva with varied pneumococcal strains. The collection of saliva can be a particularly useful in surveillance studies conducted in remote settings, as it does not require trained personnel, and DSS are resilient to various transportation conditions. MDPI 2016-03-05 /pmc/articles/PMC4813204/ /pubmed/26959014 http://dx.doi.org/10.3390/ijms17030343 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Krone, Cassandra L.
Oja, Anna E.
van de Groep, Kirsten
Sanders, Elisabeth A. M.
Bogaert, Debby
Trzciński, Krzysztof
Dried Saliva Spots: A Robust Method for Detecting Streptococcus pneumoniae Carriage by PCR
title Dried Saliva Spots: A Robust Method for Detecting Streptococcus pneumoniae Carriage by PCR
title_full Dried Saliva Spots: A Robust Method for Detecting Streptococcus pneumoniae Carriage by PCR
title_fullStr Dried Saliva Spots: A Robust Method for Detecting Streptococcus pneumoniae Carriage by PCR
title_full_unstemmed Dried Saliva Spots: A Robust Method for Detecting Streptococcus pneumoniae Carriage by PCR
title_short Dried Saliva Spots: A Robust Method for Detecting Streptococcus pneumoniae Carriage by PCR
title_sort dried saliva spots: a robust method for detecting streptococcus pneumoniae carriage by pcr
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4813204/
https://www.ncbi.nlm.nih.gov/pubmed/26959014
http://dx.doi.org/10.3390/ijms17030343
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