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In-cell western assay as a high-throughput approach for Chlamydia trachomatis quantification and susceptibility testing to antimicrobials.
- Source :
-
PloS one [PLoS One] 2021 May 11; Vol. 16 (5), pp. e0251075. Date of Electronic Publication: 2021 May 11 (Print Publication: 2021). - Publication Year :
- 2021
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Abstract
- Chlamydia trachomatis, the leading cause of bacterial sexually transmitted diseases in developed countries, with around 127 million new cases per year, is mainly responsible for urethritis and cervicitis in women, and urethritis and epididymitis in men. Most C. trachomatis infections remain asymptomatic (>50%) and, hence, untreated, leading to severe reproductive complications in both women and men, like infertility. Therefore, the detection of C. trachomatis as well as the antimicrobial susceptibility testing becomes a priority, and, along the years, several methods have been recommended, like cell culture and direct immunofluorescence (DFA) on cell cultures. Herein, we described the application of In-Cell Western assay (ICW) via Odyssey CLx as a fast, more accessible, and high-throughput platform for the quantification of C. trachomatis and the screening of anti-chlamydial drugs. As a first step, we set up a standard curve by infecting cell monolayers with 2-fold serial dilutions of C. trachomatis Elementary Body (EB) suspension. Then, different unknown C. trachomatis EB suspensions were quantified and the chlamydial susceptibility testing to erythromycin was performed, using the DFA as comparison. Our results showed a very high concordance between these two assays, as evidenced by the enumeration of chlamydial IFUs as well as the determination of erythromycin Minimum Inhibitory Concentration (MIC). In conclusion, the ICW assay may be a promising candidate as an accurate and accessible methodology for C. trachomatis antimicrobial susceptibility testing.<br />Competing Interests: The authors have declared that no competing interests exist.
- Subjects :
- Bacterial Load
Cell Line
Chlamydia Infections drug therapy
Chlamydia Infections microbiology
Feasibility Studies
Humans
Microbial Sensitivity Tests
Anti-Bacterial Agents pharmacology
Blotting, Western methods
Chlamydia Infections diagnosis
Chlamydia trachomatis drug effects
Erythromycin pharmacology
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 16
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 33974662
- Full Text :
- https://doi.org/10.1371/journal.pone.0251075