Chlamydia trachomatis intra-bacterial and total plasmid copy number in clinical urogenital samples

Abstract Chlamydia trachomatis (CT) increases its plasmid numbers when stressed, as occurs in clinical trachoma samples. Most CT tests target the plasmid to increase the test sensitivity, but some only target the chromosome. We investigated clinical urogenital samples for total plasmid copy numbers...

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Autores principales:	J. A. M. C. Dirks, K. Janssen, C. J. P. A. Hoebe, T. H. B. Geelen, M. Lucchesi, N. H. T. M. Dukers-Muijrers, P. F. G. Wolffs
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6d2731687b4046a2af2fe0945452c650
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spelling	oai:doaj.org-article:6d2731687b4046a2af2fe0945452c6502021-12-02T11:46:07ZChlamydia trachomatis intra-bacterial and total plasmid copy number in clinical urogenital samples10.1038/s41598-020-80645-y2045-2322https://doaj.org/article/6d2731687b4046a2af2fe0945452c6502021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-80645-yhttps://doaj.org/toc/2045-2322Abstract Chlamydia trachomatis (CT) increases its plasmid numbers when stressed, as occurs in clinical trachoma samples. Most CT tests target the plasmid to increase the test sensitivity, but some only target the chromosome. We investigated clinical urogenital samples for total plasmid copy numbers to assess its diagnostic value and intra-bacterial plasmid copy numbers to assess its natural variation. Both plasmid and chromosome copies were quantified using qPCR, and the plasmid:chromosome ratio (PCr) calculated in two cohorts: (1) 383 urogenital samples for the total PCR (tPCr), and (2) 42 vaginal swabs, with one half treated with propium-monoazide (PMA) to prevent the quantification of extracellular DNA and the other half untreated to allow for both tPCr and intra-bacterial PCr (iPCr) quantification. Mann–Whitney U tests compared PCr between samples, in relation to age and gender. Cohort 1: tPCr varied greatly (1–677, median 16). Median tPCr was significantly higher in urines than vaginal swabs (32 vs. 11, p < 0.001). Cohort 2: iPCr was more stable than tPCr (range 0.1–3 vs. 1–11). To conclude, tPCr in urogenital samples was much more variable than previously described. Transport time and temperature influences DNA degradation, impacting chromosomal DNA more than plasmids and urine more than vaginal samples. Data supports a plasmid target in CT screening assays to increase clinical sensitivity.J. A. M. C. DirksK. JanssenC. J. P. A. HoebeT. H. B. GeelenM. LucchesiN. H. T. M. Dukers-MuijrersP. F. G. WolffsNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-6 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q J. A. M. C. Dirks K. Janssen C. J. P. A. Hoebe T. H. B. Geelen M. Lucchesi N. H. T. M. Dukers-Muijrers P. F. G. Wolffs Chlamydia trachomatis intra-bacterial and total plasmid copy number in clinical urogenital samples
description	Abstract Chlamydia trachomatis (CT) increases its plasmid numbers when stressed, as occurs in clinical trachoma samples. Most CT tests target the plasmid to increase the test sensitivity, but some only target the chromosome. We investigated clinical urogenital samples for total plasmid copy numbers to assess its diagnostic value and intra-bacterial plasmid copy numbers to assess its natural variation. Both plasmid and chromosome copies were quantified using qPCR, and the plasmid:chromosome ratio (PCr) calculated in two cohorts: (1) 383 urogenital samples for the total PCR (tPCr), and (2) 42 vaginal swabs, with one half treated with propium-monoazide (PMA) to prevent the quantification of extracellular DNA and the other half untreated to allow for both tPCr and intra-bacterial PCr (iPCr) quantification. Mann–Whitney U tests compared PCr between samples, in relation to age and gender. Cohort 1: tPCr varied greatly (1–677, median 16). Median tPCr was significantly higher in urines than vaginal swabs (32 vs. 11, p < 0.001). Cohort 2: iPCr was more stable than tPCr (range 0.1–3 vs. 1–11). To conclude, tPCr in urogenital samples was much more variable than previously described. Transport time and temperature influences DNA degradation, impacting chromosomal DNA more than plasmids and urine more than vaginal samples. Data supports a plasmid target in CT screening assays to increase clinical sensitivity.
format	article
author	J. A. M. C. Dirks K. Janssen C. J. P. A. Hoebe T. H. B. Geelen M. Lucchesi N. H. T. M. Dukers-Muijrers P. F. G. Wolffs
author_facet	J. A. M. C. Dirks K. Janssen C. J. P. A. Hoebe T. H. B. Geelen M. Lucchesi N. H. T. M. Dukers-Muijrers P. F. G. Wolffs
author_sort	J. A. M. C. Dirks
title	Chlamydia trachomatis intra-bacterial and total plasmid copy number in clinical urogenital samples
title_short	Chlamydia trachomatis intra-bacterial and total plasmid copy number in clinical urogenital samples
title_full	Chlamydia trachomatis intra-bacterial and total plasmid copy number in clinical urogenital samples
title_fullStr	Chlamydia trachomatis intra-bacterial and total plasmid copy number in clinical urogenital samples
title_full_unstemmed	Chlamydia trachomatis intra-bacterial and total plasmid copy number in clinical urogenital samples
title_sort	chlamydia trachomatis intra-bacterial and total plasmid copy number in clinical urogenital samples
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/6d2731687b4046a2af2fe0945452c650
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Chlamydia trachomatis intra-bacterial and total plasmid copy number in clinical urogenital samples

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