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Combination with antimicrobial peptide lyses improves loop-mediated isothermal amplification based method for Chlamydia trachomatis detection directly in urine sample

BACKGROUND: Chlamydia trachomatis is an obligate intracellular human pathogen and is the most common cause of sexually transmitted diseases affecting both men and women. The pathogen can cause prostatitis and epididymitis in men. In women, cervicitis, pelvic inflammatory disease, ectopic pregnancy a...

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Autores principales: Jevtuševskaja, Jekaterina, Uusna, Julia, Andresen, Liis, Krõlov, Katrin, Laanpere, Made, Grellier, Tiia, Tulp, Indrek, Langel, Ülo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4944247/
https://www.ncbi.nlm.nih.gov/pubmed/27412444
http://dx.doi.org/10.1186/s12879-016-1674-0
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author Jevtuševskaja, Jekaterina
Uusna, Julia
Andresen, Liis
Krõlov, Katrin
Laanpere, Made
Grellier, Tiia
Tulp, Indrek
Langel, Ülo
author_facet Jevtuševskaja, Jekaterina
Uusna, Julia
Andresen, Liis
Krõlov, Katrin
Laanpere, Made
Grellier, Tiia
Tulp, Indrek
Langel, Ülo
author_sort Jevtuševskaja, Jekaterina
collection PubMed
description BACKGROUND: Chlamydia trachomatis is an obligate intracellular human pathogen and is the most common cause of sexually transmitted diseases affecting both men and women. The pathogen can cause prostatitis and epididymitis in men. In women, cervicitis, pelvic inflammatory disease, ectopic pregnancy and acute or chronic pelvic pain are frequent complications. More than half of C. trachomatis-positive patients have minimal or no symptoms, providing an ongoing reservoir for the infection. The lack of sensitive large-scale applicable point- of- care (POC) tests for C. trachomatis detection makes it difficult to diagnose chlamydia infection efficiently in resource-limited environments. METHODS: A rapid and sensitive assay based on loop-mediated isothermal amplification method (LAMP) was combined with antimicrobial peptide lysis, which is able to detect at least 7 C. trachomatis pathogens per reaction directly from urine samples. RESULTS: Our study comprising 91 first-void urine samples showed that specificity of the assay is 100 % and sensitivity 73 % when using antimicrobial peptide lysis mix. Additionally we demonstrate that our assay does not give any cross-reactivity with 30 pathogen’s DNA potentially present in the urine samples. Furthermore, the assay’s novel approach does not require purification or extraction of DNA from clinical sample prior to amplification, so the need for specialized equipment is eliminated. CONCLUSIONS: The whole procedure is significantly less laborious, less time-consuming and consequently less expensive for early detection and identification of infectious disease. C. trachomatis specific LAMP assay is relatively simple to perform and could therefore be applied in numerous POC settings. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12879-016-1674-0) contains supplementary material, which is available to authorized users.
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spelling pubmed-49442472016-07-26 Combination with antimicrobial peptide lyses improves loop-mediated isothermal amplification based method for Chlamydia trachomatis detection directly in urine sample Jevtuševskaja, Jekaterina Uusna, Julia Andresen, Liis Krõlov, Katrin Laanpere, Made Grellier, Tiia Tulp, Indrek Langel, Ülo BMC Infect Dis Research Article BACKGROUND: Chlamydia trachomatis is an obligate intracellular human pathogen and is the most common cause of sexually transmitted diseases affecting both men and women. The pathogen can cause prostatitis and epididymitis in men. In women, cervicitis, pelvic inflammatory disease, ectopic pregnancy and acute or chronic pelvic pain are frequent complications. More than half of C. trachomatis-positive patients have minimal or no symptoms, providing an ongoing reservoir for the infection. The lack of sensitive large-scale applicable point- of- care (POC) tests for C. trachomatis detection makes it difficult to diagnose chlamydia infection efficiently in resource-limited environments. METHODS: A rapid and sensitive assay based on loop-mediated isothermal amplification method (LAMP) was combined with antimicrobial peptide lysis, which is able to detect at least 7 C. trachomatis pathogens per reaction directly from urine samples. RESULTS: Our study comprising 91 first-void urine samples showed that specificity of the assay is 100 % and sensitivity 73 % when using antimicrobial peptide lysis mix. Additionally we demonstrate that our assay does not give any cross-reactivity with 30 pathogen’s DNA potentially present in the urine samples. Furthermore, the assay’s novel approach does not require purification or extraction of DNA from clinical sample prior to amplification, so the need for specialized equipment is eliminated. CONCLUSIONS: The whole procedure is significantly less laborious, less time-consuming and consequently less expensive for early detection and identification of infectious disease. C. trachomatis specific LAMP assay is relatively simple to perform and could therefore be applied in numerous POC settings. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12879-016-1674-0) contains supplementary material, which is available to authorized users. BioMed Central 2016-07-13 /pmc/articles/PMC4944247/ /pubmed/27412444 http://dx.doi.org/10.1186/s12879-016-1674-0 Text en © The Author(s). 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Jevtuševskaja, Jekaterina
Uusna, Julia
Andresen, Liis
Krõlov, Katrin
Laanpere, Made
Grellier, Tiia
Tulp, Indrek
Langel, Ülo
Combination with antimicrobial peptide lyses improves loop-mediated isothermal amplification based method for Chlamydia trachomatis detection directly in urine sample
title Combination with antimicrobial peptide lyses improves loop-mediated isothermal amplification based method for Chlamydia trachomatis detection directly in urine sample
title_full Combination with antimicrobial peptide lyses improves loop-mediated isothermal amplification based method for Chlamydia trachomatis detection directly in urine sample
title_fullStr Combination with antimicrobial peptide lyses improves loop-mediated isothermal amplification based method for Chlamydia trachomatis detection directly in urine sample
title_full_unstemmed Combination with antimicrobial peptide lyses improves loop-mediated isothermal amplification based method for Chlamydia trachomatis detection directly in urine sample
title_short Combination with antimicrobial peptide lyses improves loop-mediated isothermal amplification based method for Chlamydia trachomatis detection directly in urine sample
title_sort combination with antimicrobial peptide lyses improves loop-mediated isothermal amplification based method for chlamydia trachomatis detection directly in urine sample
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4944247/
https://www.ncbi.nlm.nih.gov/pubmed/27412444
http://dx.doi.org/10.1186/s12879-016-1674-0
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