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

English version České info

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.

Keywords:
Loop-mediated isothermal amplification, Point-of-care assay, Chlamydia trachomatis, Pathogen detection in crude urine, Diagnostics of sexually transmitted diseases

Autoři: Jekaterina Jevtuševskaja ^1*; Julia Uusna ^1,2; Liis Andresen ¹; Katrin Krõlov ¹; Made Laanpere ³; Tiia Grellier ¹; Indrek Tulp ^2,4; Ülo Langel ^1,5
Působiště autorů: University of Tartu, Institute of Technology Laboratory of Molecular Biotechnology, Nooruse 1, Tartu 50 11, Estonia. ¹; SelfD Technologie GmbH, Leipzig, Germany. ²; The Tartu University Hospital’s Women’s Clinic and Tartu Sexual Health Clinic, Tartu Estonia. ³; University of Tartu, Institute of Chemistry, Tartu, Estonia. 5Stockholm University, Stockholm, Sweden. ⁴
Vyšlo v časopise: BMC Infectious diseases 2016, 16:329
Kategorie: Research article
prolekare.web.journal.doi_sk: https://doi.org/10.1186/s12879-016-1674-0

© 2016 The Author(s).
Open Access This 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.
The electronic version of this article is the complete one and can be found online at: http://bmcinfectdis.biomedcentral.com/articles/10.1186/s12879-016-1674-0

Souhrn

Keywords:
Loop-mediated isothermal amplification, Point-of-care assay, Chlamydia trachomatis, Pathogen detection in crude urine, Diagnostics of sexually transmitted diseases

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