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      Polymerase Chain Reaction as a Diagnostic Tool for Six Sexually Transmitted Infections - Preliminary Results

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          Abstract

          Background and aim

          Sexually transmitted infections are a very frequent and under-diagnosed cause of illness worldwide. A high number of detection methods and a large range of specimens in which sexually transmitted infections can be determined are available at the moment. Polymerase chain reaction performed on first void urine offers the advantage of being non-invasive, self-collectable and has high sensitivity and specificity. We looked to determine the frequency of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Mycoplasma hominis, Mycoplasma genitalium and Ureaplasma urealyticum in symptomatic and asymptomatic patients.

          Methods

          Six sexually transmitted infections were determined in the first void urine of 15 symptomatic and asymptomatic patients by polymerase chain reaction. We used “Epicenter MasterPure™ Complete DNA and RNA Purification Kit” for the DNA purification and “Seeplex® STD6 ACE Detection” for the DNA amplification. The results were examined in UV light.

          Results

          A number of 5 patients had positive results for Chlamydia trachomatis or Neisseria gonorrhoeae. Sexually transmitted infections are more frequent in men between 27 and 40 years old.

          Conclusions

          Polymerase chain reaction is a good diagnostic tool for sexually transmitted infections because it has a high sensitivity and specificity. Chlamydia trachomatis is the most frequent sexually transmitted infection, followed by Neisseria gonorrhoeae.

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          Most cited references23

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          Sensitive and rapid detection of Chlamydia trachomatis by recombinase polymerase amplification directly from urine samples.

          Katrin Krõlov, Jekaterina Frolova, Oana M. Tudoran … (2014)
          Chlamydia trachomatis is the most common sexually transmitted human pathogen. Infection results in minimal to no symptoms in approximately two-thirds of women and therefore often goes undiagnosed. C. trachomatis infections are a major public health concern because of the potential severe long-term consequences, including an increased risk of ectopic pregnancy, chronic pelvic pain, and infertility. To date, several point-of-care tests have been developed for C. trachomatis diagnostics. Although many of them are fast and specific, they lack the required sensitivity for large-scale application. We describe a rapid and sensitive form of detection directly from urine samples. The assay uses recombinase polymerase amplification and has a minimum detection limit of 5 to 12 pathogens per test. Furthermore, it enables detection within 20 minutes directly from urine samples without DNA purification before the amplification reaction. Initial analysis of the assay from clinical patient samples had a specificity of 100% (95% CI, 92%-100%) and a sensitivity of 83% (95% CI, 51%-97%). The whole procedure is fairly simple and does not require specific machinery, making it potentially applicable in point-of-care settings. Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
          Article 0 comments Cited 55 times – based on 0 reviews     Review now
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            Assessment of Chlamydia trachomatis, Ureaplasma urealyticum, Ureaplasma parvum, Mycoplasma hominis, and Mycoplasma genitalium in semen and first void urine specimens of asymptomatic male partners of infertile couples.

            R Gdoura, W Kchaou, L Ammar-Keskes … (2015)
            The purpose of this study was threefold: to compare semen and first void urine (FVU) specimens from asymptomatic infertile men for the detection of Chlamydia trachomatis, genital ureaplasma, and genital mycoplasma infections using in-house inhibitor-controlled polymerase chain reaction (PCR)-microtiter plate hybridization assay; to determine the prevalence of those organisms in infertile men in Tunisia; and to study the relationship between these bacteria and male infertility. Paired urine and semen specimens from 104 patients were examined by in-house PCR for the presence of DNA of Chlamydia trachomatis, genital ureaplasmas (Ureaplasma urealyticum and Ureaplasma parvum) and genital mycoplasmas (Mycoplasma hominis and Mycoplasma genitalium). Semen analysis was assessed according to the guidelines of the World Health Organization. Nominal scale variables, the Mann-Whitney test, and the Kruskal-Wallis nonparametric analysis of variance test were used for statistical analysis. There was a very high concordance (>95%) and a very good agreement (kappa > 0.9) between the detection of Chlamydia trachomatis, genital ureaplasmas, and Mycoplasma hominis in semen and corresponding FVU specimens. Our findings also show a high concordance (81.1%) and a good agreement (kappa = 0.79) between the detection of Mycoplasma genitalium in both specimens. C trachomatis, genital mycoplasmas, and genital ureaplasmas were found to be widespread among infertile male patients in Tunisia, as shown by their respective prevalences of 43.3%, 18.3%, and 14.4%. The mean values of seminal volume, sperm concentration, sperm viability, sperm motility, sperm morphology, and leukocyte count were not significantly related either to the detection of C trachomatis DNA or to that of genital ureaplasma or mycoplasma DNA in semen specimens. Using our in-house PCR, both semen and FVU were found to be sensitive diagnostic specimens for the detection of C trachomatis, ureaplasmas, and mycoplasmas. The FVU, a less invasive and self-collected specimen, can serve as a marker for the presence of these organisms in the genital tract and can be used as a reliable way of detecting asymptomatic carriers of infection.
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              Performance of Anyplex™ II multiplex real-time PCR for the diagnosis of seven sexually transmitted infections: comparison with currently available methods.

              Dong Sup Lee, Seung-Ju Lee, Sung-Hoo Hong … (2013)
              The real-time PCR assay is the most sensitive test for screening and diagnosing sexually transmitted infections (STIs) and has made diagnosing these infections easier for clinicians. The aim of this study was to investigate the reliability, accuracy, and usefulness of the real-time multiplex PCR assay for the detection of seven sexually transmitted microorganisms in clinical samples.
              Article 0 comments Cited 36 times – based on 0 reviews     Review now
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                Author and article information

                Journal
                Journal ID (nlm-ta): Clujul Med
                Journal ID (iso-abbrev): Clujul Med
                Journal ID (publisher-id): CM
                Title: Clujul Medical
                Publisher: Iuliu Hatieganu University of Medicine and Pharmacy
                ISSN (Print): 1222-2119
                ISSN (Electronic): 2066-8872
                Publication date (Print): 2015
                Publication date (Electronic): 28 January 2015
                Volume: 88
                Issue: 1
                Pages: 33-37
                Affiliations
                [1 ]Department of Dermatology, University of Medicine and Pharmacy Iuliu Hatieganu, Cluj Napoca, Romania
                [2 ]Department of Cellular and Mollecular Biology, University of Medicine and Pharmacy Iuliu Hatieganu, Cluj Napoca, Romania
                [3 ]Department of Urology, Clinical Municipal Hospital, Cluj Napoca, Romania
                [4 ]Department of Urology, University of Medicine and Pharmacy Iuliu Hatieganu, Cluj Napoca, Romania
                Author notes
                Address for correspondence: demian.alecsandra@ 123456umfcluj.ro
                Article
                Publisher ID: cm8801p33
                DOI: 10.15386/cjmed-373
                PMC ID: 4508611
                PubMed ID: 26528045
                SO-VID: 097b222b-99d1-4cd8-8bde-0ede9f8862b7
                Copyright statement: Copyright @ 2015
                License:

                This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License

                History
                Date received : 30 November 2014
                Date revision received : 18 December 2014
                Date accepted : 26 December 2014
                Categories
                Subject: Original Research
                Subject: Dermatology

                Keywords: sexually transmitted infections,urine,polymerase chain reaction
                Data availability:
                Keywords: sexually transmitted infections, urine, polymerase chain reaction

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