Background Sexually transmitted infections (STIs) are prevalent throughout the world and impose a significant burden on individual health and public health systems. Missed diagnosis and late treatment of STIs can lead to serious complications such as infertility and cervical cancer. Although sexually transmitted co-infections are common, most commercial assays target one or a few STIs. The HPV-STI ChapterDx Next Generation Sequencing (NGS) assay detects and quantifies 29 HPVs and 14 other STIs in a single-tube and single-step PCR reaction and can be applied to tens to thousands of samples in a single sequencing run. Methods A cohort of 274 samples, previously analyzed by conventional cytology/histology and Roche cobas HPV Test, were analyzed by ChapterDx HPV-STI NGS assay for detection of 43 HPV and STI. A set of 43 synthetic control DNA fragments for 43 HPV and STI were developed to evaluate the limit of detection, specificity, and sensitivity of ChapterDx HPV-STI NGS assay. Results The assay was evaluated in this study, and the limit of detection was 100% at 50 copies for all targets, and 100%, 96%, 88% at 20 copies for 34, 8, and 1 target, respectively. The performance of this assay has been compared to Roche cobas HPV test, showing an overall agreement of 97.5% for hr-HPV, and 98.5% for both, HPV16 and HPV18. The assay also detected all HPV-infected CIN2/3 with 100% agreement with Roche cobas HPV results. Moreover, several co-infections with non-HPV STIs, such as C. trachomatis, T. vaginalis, M. genitalium, and HSV2 were identified. Conclusions The ChapterDx HPV-STI NGS assay is a user-friendly, easy to automate and cost-efficient assay, which provides accurate and comprehensive results for a wide spectrum of HPVs and STIs.
Introduction: Mycoplasma genitalium (MG) is responsible for 15%-20% nongonococcal urethritis in men. In Argentina, the diagnosis is only performed by few laboratories. Single-dose 1 g azithromycin (AZM1D) treatment leads to emergence of macrolide resistance (mutations at 23S rRNA gene, region V, position 2058 or 2059). Recommendations include 5-day AZM (AZM5D) regimen, moxifloxacin as second-line therapy. Doxycycline is only 30% effective. Test of Cure (ToC) is advisable. Objective: The aim of this study was to describe the first two clinical cases of persistent and recurrent urethritis due to macrolide-resistant MG in Argentina. Methods: End point polymerase chain reaction (PCR) for diagnosis and ToC. Sanger sequencing analysis of mutations. Results: Case 1: A 26-year-old male patient with occasional heterosexual contacts and no history of sexually transmitted infections (STIs) complained urethral thick purulent discharge and dysuria (January 2018), with negative microbiological cultures and Chlamydia trachomatis PCR. The patient received ceftriaxone/AZM1D. However, symptoms persisted (April 2018). Later, doxycycline was prescribed for 1 month. Five days after treatment, the sample was referred to the STI national reference laboratory (NRL) and results were found positive for MG. The patient was given AZM5D. As a result, symptoms disappeared, posterior ToC was found negative, and retrospectively, sequencing 23S rRNA gene showed A2058G transition. Case 2: An 18-year-old male patient with stable heterosexual relationship complained of previous gonococcal urethritis and urethral serous exudate with inflammatory reaction (September 2017), with negative microbiological cultures. The patient received ceftriaxone and AZM1D as initial treatment. Later, he was given doxycycline for 10 days. On February 2018, symptoms reappeared and sample referred to the NRL was positive for MG (negative for other STIs). With AZM1D treatment, symptoms disappeared. After 1 month, the symptoms recurred. Results showed a new MG-positive sample (April 2018). AZM5D administration induced 2 weeks symptoms free and recurrence, requiring moxifloxacin treatment. Symptoms disappeared completely. Posterior ToC is negative. Subsequently, sequencing both samples referred to the NRL showed A2059G transition. Conclusion: The clinical cases presented notified the importance of early and accurate diagnosis of MG infections and use of adequate treatment schemes. We emphasized the relevance of monitoring and surveillance prevalence of macrolide-resistant MG in Argentina.
Mg 23s rRNA gene 27 different urogenital samples were analyzed (Fig. 1). Median age of patients was 25 years (a range 18 to 47). 19 patients were symptomatic at the time of sampling and 5 presented Chlamydia trachomatis co-infection (Fig. 2). Of the 27 Mg positive only 22 could be sequenced in search for mutations. 27 Mg positive samples recovered from genital sites submitted for diagnosis to the National Reference Laboratory for Sexual Transmitted Diseases between 2013-2017 were analyzed.Mycoplasma genitalium (Mg) is a sexually transmitted pathogen responsible for approximately 15%-20% of nongonococcal urethritis in male. In women several studies have shown an association between Mg and cervicitis, endometritis and pelvic inflammatory disease, although the association between infection and symptoms is weaker compared to men. In recent years, an increase in the Mg resistance to several antimicrobial agents used in treatments has been observed. Thus, it is speculated that Mg is evolving into what is commonly classified as superbug.
Sexually transmitted infections (STIs) are prevalent throughout the world and impose a significant burden on individual health and public health systems. Missed diagnosis and late treatment of STIs can lead to serious complications such as infertility and cervical cancer. Although sexually transmitted co-infections are common, most commercial assays target one or a few STIs. The HPV-STI ChapterDx Next Generation Sequencing (NGS) assay detects and quantifies 29 HPVs and 14 other STIs in a single-tube and single-step PCR reaction and can be applied to tens to thousands of samples in a single sequencing run. The assay was evaluated in this study, and the limit of detection was 100% at 50 copies for all targets, and 100%, 96%, 88% at 20 copies for 34, 8, and 1 target, respectively. The performance of this assay has been compared to Roche cobas HPV test, showing an overall agreement of 97.5% for hr-HPV, and 98.5% for both, HPV16 and HPV18. The assay also detected all HPV-infected CIN2/3 with 100% agreement with Roche cobas HPV results. Moreover, several co-infections with non-HPV STIs, such as C. trachomatis, T. vaginalis, M. genitalium, and HSV2 were identified. The ChapterDx HPV-STI NGS assay is a user-friendly, easy to automate and cost-efficient assay, which provides accurate and comprehensive results for a wide spectrum of HPVs and STIs.
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