Journal article
Detection of Chlamydia trachomatis mRNA using digital PCR as a more accurate marker of viable organism
European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology, Vol.37(11), pp.2117-2122
2018
Abstract
Spontaneous resolution of urogenital Chlamydia trachomatis (CT) without treatment has previously been described, but a limitation of these reports is that DNA or RNA-based amplification tests used do not differentiate between viable infection and non-viable DNA. We modified a previously published CT mRNA detection (omp2) method to differentiate between viable infection and non-viable DNA in a sample of CT DNA PCR positive women. We modified a CT mRNA detection (omp2) method from reverse transcriptase qPCR (RTqPCR) to digital PCR (dPCR) and evaluated it in samples from CT DNA positive women. Firstly, CT infected McCoy B cells treated with azithromycin in vitro identified detectable mRNA levels disappeared <2 days, while DNA persisted up to 6 days. We used 55 self-collected vaginal swabs from a cohort of women diagnosed as DNA positive for chlamydia obtained pre- and 7 days of post-azithromycin treatment. Concordance with DNA results was higher for dPCR than RTqPCR (74.5% versus 65.5%). At visit 1, there was a strong linear relationship between DNA and mRNA (r = 0.9, p < 0.000); 24 samples had both mRNA and DNA detected (82.8%) and 5 had only DNA detected with a potential false positive proportion of 17.2% (95%CI: 5.8, 35.8). At visit 2, there was poor correlation between DNA and mRNA (r = 0.14, p = 0.55); eight specimens had only DNA detected (42.1%; 95%CI: 20.25, 66.50) and one had mRNA detected. DNA detection methods alone may detect non-viable DNA. Consideration should be given to further develop mRNA assays as ancillary tests to improve detection of viable chlamydia.
Details
- Title
- Detection of Chlamydia trachomatis mRNA using digital PCR as a more accurate marker of viable organism
- Authors
- Samuel Phillips (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and EngineeringLenka A Vodstrcil (Author) - University of MelbourneWilhelmina M Huston (Author) - University of Technology SydneyAmba Lawerence (Author) - Queensland University of TechnologyPeter Timms (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and EngineeringMarcus Y Chen (Author) - Melbourne Sexual Health CentreKaren Worthington (Author) - Melbourne Sexual Health CentreRuthy McIver (Author) - Sydney Sexual Health CentreCatriona S Bradshaw (Author) - Melbourne Sexual Health CentreSuzanne M Garland (Author) - University of MelbourneSepehr N Tabrizi (Author) - University of MelbourneJane S Hocking (Author) - University of Melbourne
- Publication details
- European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology, Vol.37(11), pp.2117-2122
- Publisher
- Springer
- Date published
- 2018
- DOI
- 10.1007/s10096-018-3347-y
- ISSN
- 0934-9723
- Organisation Unit
- University of the Sunshine Coast, Queensland; School of Health and Sport Sciences - Legacy; School of Science, Technology and Engineering; Centre for Bioinnovation
- Language
- English
- Record Identifier
- 99450731002621
- Output Type
- Journal article
Metrics
2 File views/ downloads
396 Record Views
InCites Highlights
These are selected metrics from InCites Benchmarking & Analytics tool, related to this output
- Collaboration types
- Domestic collaboration
- Web Of Science research areas
- Infectious Diseases
- Microbiology
UN Sustainable Development Goals (SDGs)
This output has contributed to the advancement of the following goals:
Source: InCites