Neisseria Gonorrhoeae Research Articles

Antimicrobial Resistance of Neisseria gonorrhoeae Isolates among Men Who Have Sex with Men in Lower Silesia, Poland.

Neisseria gonorrhoeae (NG) has developed resistance to nearly all antibiotics used for its treatment. However, very limited data are available regarding the antimicrobial resistance of NG isolates among MSM in Poland. The aim of this study was to evaluate the susceptibility of Neisseria gonorrhoeae isolates in this key population. We investigated the antimicrobial susceptibility of NG isolates to six antimicrobials (ceftriaxone, cefixime, azithromycin, ciprofloxacin, tetracycline, and benzylpenicillin). Minimum inhibitory concentrations (MICs; mg/L) were determined using Etests on gonococcal isolates. One hundred high-risk MSM were included in the study (25 HIV-positive and 75 HIV-negative using pre-exposure prophylaxis for HIV). The rate of NG infection was 28%. All NG isolates were susceptible to cefixime and ceftriaxone. Susceptibility to azithromycin was found in 69.2% (18/26) of the NG isolates and resistance in 30.8% (8/26) of NG isolates. Susceptibility to tetracycline was found in 50% (13/26) of the isolates and resistance in 50% (13/26) of the isolates. We observed gonorrhea to be more prevalent in patients with a higher number of oral sexual contacts. Increasing azithromycin resistance is especially concerning for future treatment options, especially if ceftriaxone/cefixime resistance starts to develop and for people with beta-lactam antibiotics allergies. Doxy-PEP might lose its partial efficacy for NG soon.

Zap the clap with DNA: a novel microbicide for preventing Neisseria gonorrhoeae infection.

Each year, Neisseria gonorrhoeae (Ngo) causes over 1.5 million new infections in the United States, and >87 million worldwide. The absence of a vaccine for preventing gonorrhea, the rapid emergence of multidrug-resistant and extremely drug-resistant Ngo strains, and the limited number of antibiotics available for treating gonorrhea underscore the importance of developing new modalities for addressing Ngo infection. Here, we describe DNA-based microbicides that kill Ngo but not commensals. Previously, we showed that Ngo is killed when it takes up differentially methylated DNA with homology to its genome. We exploited this Achilles heel to develop a new class of microbicides for preventing Ngo infection. These microbicides consist of DNA molecules with specific sequences and a methylation pattern different from Ngo DNA. These DNAs kill low-passage and antibiotic-resistant clinical isolates with high efficiency but leave commensals unharmed. Equally important, the DNAs are equally effective against Ngo whether they are in buffered media or personal lubricants. These findings illustrate the potential of this new class of practical, low-cost, self-administered DNA-based microbicides for preventing Ngo transmission during sexual intercourse.

Effect of antenatal Chlamydia trachomatis and Neisseria gonorrhoeae screening on postdelivery prevalence and vertical transmission in Gaborone, Botswana: findings from an exploratory study

ObjectivesChlamydia trachomatis and Neisseria gonorrhoeae are common sexually transmitted infections (STIs). Untreated infection in pregnancy can result in adverse neonatal outcomes, including vertical transmission. Screening for these infections is not...

Refinement and optimisation of Neisseria gonorrhoeae NHBA and MetQ vaccine candidates

Neisseria gonorrhoeae has a significant impact on reproductive health with an estimated 82 million new cases of infection per year worldwide. Due to the ongoing emergence of multidrug-resistant N. gonorrhoeae strains, the high number of asymptomatic cases, and the risk of disease sequelae, the development of a gonococcal vaccine is urgently needed. We have previously described two potential gonococcal vaccine antigens, cNHBA (C-terminal fragment of the Neisseria Heparin Binding Antigen) and MetQ (methionine-binding protein). This study aimed to optimise these antigens for improved immune responses and to facilitate vaccine production, by investigating cNHBA fusions with the full-length MetQ protein or N-terminal and C-terminal MetQ fragments (Met1 and Met2, respectively) adjuvanted with aluminium hydroxide. The cNHBA and MetQ fragments and fusion antigens were all immunogenic in mice, generating a predominantly IgG1 response. Antibodies mediated bacterial killing via both serum bactericidal activity (SBA) and opsonophagocytic activity (OPA), and reduced adherence to cervical and urethral epithelial cells. Among the antigen fusions tested, MetQ-cNHBA and cNHBA-Met2 generated the highest SBA, OPA and adherence blocking titres and are proposed as promising optimised antigens for N. gonorrhoeae vaccine development.

Whole-genome sequencing resolves biochemical misidentification of Neisseria species from urogenital specimens.

Neisseria meningitidis (Nm) and Neisseria gonorrhoeae (Ng) are human pathogens that sometimes occupy the same anatomical niche. Ng, the causative agent of gonorrhea, infects 87 million individuals annually worldwide and is an urgent threat due to increasing drug resistance. Ng is a pathogen of the urogenital tract and may infect the oropharyngeal or rectal site, often asymptomatically. Conversely, Nm is an opportunistic pathogen. While often a commensal in the oropharyngeal tract, it is also the leading cause of bacterial meningitis with 1.2 million cases globally, causing significant morbidity and mortality. Horizontal gene transfer (HGT) is likely to occur between Ng and Nm due to their shared anatomical niches and genetic similarity, which poses challenges for accurate detection and treatment. Routine surveillance through the Gonococcal Isolate Surveillance Project and Strengthening the U.S. Response to Resistant Gonorrhea detected six concerning urogenital Neisseria isolates with contradicting species identification in Milwaukee (MIL). While all six isolates were positive for Ng using nucleic acid amplification testing (NAAT) and matrix-assisted laser desorption/ionization time of flight identified the isolates as Ng, two biochemical tests, Gonochek-II and API NH, classified them as Nm. To address this discrepancy, we performed whole-genome sequencing (WGS) using Illumina MiSeq on all isolates and employed various bioinformatics tools. Species detection analysis using BMScan, which uses WGS data, identified all isolates as Ng. Furthermore, Kraken revealed over 98% of WGS reads mapped to the Ng genome and <1% to Nm. Recombination analysis identified putative HGT in all MIL isolates within the γ-glutamyl transpeptidase (ggt) gene, a key component in the biochemical tests used to differentiate between Nm and Ng. Further analysis identified Nm as the source of HGT event. Specifically, the active Nm ggt gene replaced the Ng pseudogenes, ggt1 and ggt2. Together, this study demonstrates that closely related Neisseria species sharing a niche underwent HGT, which led to the misidentification of species following biochemical testing. Importantly, NAAT accurately detected Ng. The misidentification highlights the importance of using WGS to continually evaluate diagnostic or bacterial identification tests.

A-359 Clinical Performance Evaluation of the Polymerase Chain Reaction (PCR)-Based cobas CT/NG/MG Test for Use on the cobas liat System in a Clinical Laboratory Setting and Point-of-Care (POC) Location

Abstract Background Screening for Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Mycoplasma genitalium (MG) can be achieved rapidly (in approximately 20 minutes) with the cobas® CT/NG/MG test (assay not cleared by US FDA. Submission currently under review and subject to change per health authority feedback). This automated, qualitative, real-time PCR-based nucleic acid amplification test (NAAT) is for use on the cobas® liat® system. This study evaluated the test’s clinical performance using urogenital samples from symptomatic and asymptomatic patients. Methods This non-interventional, non-observational study used prospective clinician-collected and self-collected specimens (urine, and vaginal swabs, all in cobas® PCR Media) from symptomatic/asymptomatic patients ≥14 years old from 13 POC sites across the US. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of cobas liat CT/NG/MG were calculated with respect to patient infected status (PIS), as determined using results from three FDA-approved NAATs and one laboratory-developed test. Due to insufficient positive NG samples during the trial period, further testing used archived samples. Results The median (range) age of the study population (N=4,800) was 35.0 (15.0-81.0) years; 40.4% (n=1,941) were symptomatic and 51.9% (n=2,489) were assigned female at birth. The cobas CT/NG/MG nucleic acid test demonstrated good clinical performance (Table 1). Across all specimen types, specificity was &amp;gt;97% for each analyte. Sensitivity was ≥95%, except in female urine (CT 87.0%, NG 93.1%, MG 78.9%). Regardless of specimen type, PPV and NPV were ≥95% for CT and NG; PPV for MG was highest in male urine (92.7%) and NPV was &amp;gt;97.5% across analytes. Conclusions In a clinical setting, the cobas CT/NG/MG nucleic acid test showed good clinical performance for the detection of CT, NG, and MG in urogenital samples, in addition to providing a short turn-around time and centralized testing lab quality at the POC for both self- and clinician-collected samples.

A-238 Lyophilized Bead Multiplex Assays for CT/NG/TV Detection

Abstract Background Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Trichomonas vaginalis (TV) cause the three most prevalent sexually transmitted infectious diseases in developed countries, contributing to approximately 300 million new infections worldwide, annually. Due to the challenges of culturing and using immunoassays to detect these pathogens, DNA-based molecular tests have been instrumental, and current trends in molecular testing for CT/NG/TV have moved toward point-of-care tests. Lyophilization provides benefits required for point-of-care tests, such as ambient storage, extended shelf life, simplified downstream workflow, and variable formats. A lyophilized-bead format enables an assay to be placed in sealed consumables and stored at room temperature for up to two years. An additional consideration is the need for endogenous and exogenous controls. Exogenous in-process controls can be used to validate the performance of workflows culminating in a qPCR reaction and provide flexibility to test developers. An ideal exogenous in-process control is nonhomologous to human or human pathogen genome sequences and does not align with primers or probes designed for pathogen detection. The objective of the current study was to develop a lyophilized-bead-format assay containing an ideal exogenous control and establish whether it is a valid option for the accurate detection of CT, NG, and TV. Methods A synthetic DNA control containing targets for CT/NG/TV/RNase P (RP)/exogenous control was used to establish an analytical LOD by qPCR. The 5-plex assay, lyophilized in beads, was hydrated and tested for linearity over an 8-log range of the synthetic DNA control. Further, the qPCR lyophilized bead with assay was tested with a proficiency sample by extracting DNA from ZeptoMetrix NATtrol CT/NG/TV Positive Control and spiking-in the exogenous DNA control. qPCR was performed on the extracted DNA and human cDNA. Results The analytical LOD for the control DNA was determined to be 5 copies/reaction. Additionally, linearity was observed by qPCR over an 8-log range of the synthetic DNA control for all five targets in multiplex with good efficiencies and R2 values. Finally, all targets were amplified and detected by qPCR using DNA extracted from ZeptoMetrix NATtrol CT/NG/TV Positive Control with spiked-in exogenous DNA control. The detection of the exogenous sequence in the qPCR multiplex reaction indicated the extraction was successful. Conclusions We have developed a multiplex qPCR assay in lyophilized-bead format that can identify infectious agents, such as CT/NG/TV, in a single reaction. We have included RP as a human host control, plus an exogenous in-process control, for a total of 5-plex in separate fluorescent channels. We have shown that our lyophilized beads with assay produce a positive qPCR result when combined with a proficiency sample.

A-247 Performance and Compatibility Assessment of Serially Diluted Microbix QAPTM Controls with Seegene NovaplexTM Assays

Abstract Background Choosing high-quality external (third-party) controls like Microbix QAPTM PROCEEDxTM Controls and REDxTM Controls is crucial during test development, validation of Laboratory Developed Tests (LDTs), performance verification, proficiency testing, instrument qualifications, operator training and R&amp;D testing. Laboratories may prefer to include multiple positive controls at varying concentrations to achieve reliable performance across a broad range of detection concentrations. In this study, we assess the performance and compatibility of Microbix QAPTM PROCEEDxTM liquid Controls and REDxTM swabs with Seegene’s real time qPCR assays that utilize TOCE and MuDTTM technologies. TOCE and MuDTTM technologies achieve multiple reads in a single channel and are the backbone of Seegene’s wide coverage panel assays. Methods The controls below were reconstituted and/or diluted in Copan UTM and extracted on the Seegene STARlet liquid handler using Seegene’s STARmag Universal extraction kit. After extraction, PCR was run on the Bio-Rad CFX 96 Opus Dx and analyzed using the Seegene Viewer software. Results Microbix QAPsTM liquid formulations were tested at various dilutions from stock up to 1/2048 with N=5per dilution. The VP-13-01 Flu A control was detected up to a concentration of1/2048 and at this dilution had Ct values of 30 + 0.3 for the Novaplex Respiratory Panel 1A (RP1A) Assay, 28.6 + 0.3 cycles for the Seegene SARS-CoV-2/FluA/FluB/RSV (Quad) Assay, and 27.9 + 0.3 for the Respiratory Virus Master (RVM) Assay. The Microbix VP-14-01 Flu B control was detected up to 1/2048 with Ct values of 34.3 + 0.3 for the RP1A Assay, 30 + 0.5 for the Quad Assay, and 30.3 + 0.3 for the RVM Assay. The VP-07-01 RSV A control was also detected at a 1/2048 dilution with Ct values of 32.7 + 0.3 for RP1A, 30.6 + 0.6 for Quad Assay, and 29.2 + 0.4 for RVM. The VP-61-01 Trichomonas vaginalis (TV) control was detected up to a dilution of 1/128 with a Ct value of 37.3 + 0.4 when tested with the Seegene Sexually Transmitted Infection Essential (STI) Assay. The VP-02-M1 HSV1 control was detected up to a dilution of 1/128 with a Ct value of 31.3 + 0.7 for the Seegene Genital Ulcer (GU) Assay. The VP-23-M1 HSV2 control was detected p to a dilution of 1/128 with a Ct value of 31.9 + 0.5 for the GU Assay. Microbix also makes a dry swab formulation product line that we tested in 3mL of UTM. Of these we tested the RED-S-10-M1 swab covering the SARS-CoV-2, Flu A, Flu B, and RSV controls. The VP-S-12-M5 swab covering the Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Mycoplasma genitalium (MG), and TV controls. We also tested the VP-S-03-M1 swab covering HSV1, HSV2, VZV and syphilis controls. When tested with various assays all swabs were detected at the stock formulation. Conclusions Microbix QAPsTM are inactivated whole-organism controls, mimicking patient samples, and meet the stringent demand for quality or performance assessments. Their compatibility with Seegene’s Assays over a range of dilutions and formulations makes them ideal for ensuring reliable detection across multiple diagnostic needs.

A-296 Streck Urine Preserve Provides Sample Stability for Cepheid Xpert CT/NG test

Abstract Background Sexually transmitted infections (STIs) are one of the most common communicable diseases worldwide and are associated with significant morbidity and mortality. Data indicates that the four curable STIs - chlamydia, gonorrhea, trichomoniasis, and syphilis - cause over 375 million infections annually. Further, the infection rate for gonorrhea has increased by 63% in the United States and the emergence of resistant gonorrhea is an urgent global public health concern. To prevent transmission of gonorrhea and other curable STIs and minimize the escalation of untreated infections, early and effective prevention and control strategies are paramount. Rapid diagnostic testing and nucleic acid amplification tests (NAATs) have become the gold standard for STI detection. However, limited global access to these technologies often requires that samples be shipped to central laboratories for analysis. Current protocols recommend that neat urine be analyzed within 24 hours of collection if stored at 2 °C to 8 °C or within four days if frozen, as prolonged storage and exposure to freeze-thaws can induce cell lysis and release of nucleases, leading to degradation of DNA and false negative test results. Without sample stabilization, laboratories are left with a small window for accurate analysis. Here, we demonstrate that Streck Urine Preserve (SUP) maintains target and donor DNA concentrations during prolonged storage and that these stabilized samples are compatible with Cepheid’s automated GeneXpert System and associated Xpert CT/NG test. Methods Fresh human urine, with or without SUP, was tested neat or spiked with Neisseria gonorrhoeae (N. gonorrhoeae). Samples were stored at room temperature for up to 18 days before being analyzed on the Cepheid GeneXpert System using the Xpert CT/NG test per manufacturer’s instructions. Differences in average Ct values were compared for the sample processing control, sample adequacy control, and N. gonorrhoeae targets. DNA stability was assessed by comparing the changes between fresh and SUP-treated samples throughout the study period. Results Following prolonged storage, Ct values for neat, untreated urine increased by 3-7 cycles, correlating to a 10- to 100-fold decrease in detectable copies of N. gonorrhoeae and the sample adequacy control compared to urine samples stabilized with SUP. The average Ct value for samples stabilized with SUP did not change by more than 1.5 cycles throughout the study period. Additionally, Ct values for the sample processing control remained constant throughout the testing period, indicating that SUP did not impact sample processing. Conclusions When limited point-of-care testing options require storage and shipping of samples to central laboratories, urine stabilizers are imperative to obtain results representative of the patient sample at the time of collection. SUP effectively stabilized N. gonorrhoeae DNA and donor DNA prior to analysis with the Cepheid Xpert CT/NG test. This contrasts with the marked decrease in DNA copy number in non-stabilized urine over the study duration, which may lead to false negative results for low-positive urine samples. Taken together, SUP provides clinical researchers and assay developers with added flexibility when handling, shipping, and processing urine specimens and supports compatibility with automated NAAT systems and tests.

A-301 Evaluation of Second-Generation Visby Medical Sexual Health Point-of-Care Test: Clinical Concordance, Analytical Error Rates, and Result Turnaround Time

Abstract Background Rapid testing for Neisseria gonorrhoeae, Chlamydia trachomatis, and Trichomonas vaginalis may be useful for antibiotic stewardship initiatives. Our institution recently implemented the second-generation Visby Medical Sexual Health Test (Visby Gen2) for rapid diagnosis of STIs in our emergency department. Here we (1) compared result concordance of the (Visby Gen2) to the first-generation Visby Medical Sexual Health Click Test (Visby Gen1), (2) compared assay failure rates of the Visby Gen1 to Gen2 tests, and (3) compared result turnaround time (TAT) for clinically ordered rapid STI tests to central laboratory STI results. Methods Accuracy was assessed using residual clinical samples in Aptima buffer submitted for routine molecular STI testing (C. trachomatis, N. gonorrhoeae, and T. vaginalis) in the central laboratory on the Panther platform (Hologic). Aptima samples were diluted 1:33 in Visby collection buffer prior to testing on the Visby cartridge. Clinically reported results were used as reference results. Diluted samples with discordant results were re-tested on the reference method and the result from the dilution was used in calculations. Assay failure rates were calculated as the number of failed assays compared to the total assays performed. TAT for rapid and central lab testing was calculated as time of sample received in laboratory to time verified in laboratory information system. Results For Visby Gen1 we observed negative concordance of 100% (41/41) for C. trachomatis, 97.5% (39/40) for N. gonorrhoeae, and 97.1% (34/35) for T. vaginalis. Positive concordance was 91.7% (11/12) for C. trachomatis, 100% (11/11) for N. gonorrhoeae, and 100% (17/17) for T. vaginalis. For Visby Gen1 we observed a 10.5% (12/114) cartridge failure rate. For Visby Gen2, we observed negative concordance of 100% (59/59) for C. trachomatis, 100% (63/63) for N. gonorrhoeae, and 100% (55/55) for T. vaginalis while positive concordance was 100% (10/10) for C. trachomatis, 100% (6/6) for N. gonorrhoeae, and 90.9% (10/11) for T. vaginalis. The Visby Gen2 assay displayed a 4.9% (4/81) cartridge failure rate. Six weeks after implementing the Visby Gen2 assay in our ED, rapid STI panel TAT was 0.48 hours on average (n=20). In the central laboratory the average TAT was 22.31 hours (n=7892). Conclusions Assay concordance to reference method was similar for Visby Gen1 and Gen2 assays. The cartridge failure rate improved by 46.7% with the Gen2 assay which should decrease the need to repeat testing due to assay failure. Result TAT decreased by 97.8% when comparing rapid Visby results to central laboratory. Therefore, Visby STI testing using the Gen2 assay can provide accuracy and rapid results likely to facilitate more rapid patient management.

Emergence and evolution of mosaic penA-60 and penA-237 alleles in a Neisseria gonorrhoeae core genogroup that was historically susceptible to extended spectrum cephalosporins.

Neisseria gonorrhoeae (Ng) has successively developed resistance to all previously recommended antimicrobial therapies, with ceftriaxone being the last option for monotherapy of gonorrhea. Global emergence and international spread of the FC428 clone derived mosaic penA-60 allele, associated with highlevel ceftriaxone minimum inhibitory concentrations (MICs) in non FC428 clone Ng lineages, has become an increasing concern. The penA-60 allele carrying Ng was first identified in the U.S. in Las Vegas, Nevada (2019; GCWGS-102723), with a multi-locus sequence type (MLST)-1901 strain, in a non FC428 clone Ng lineage, which is associated with a historically ceftriaxone susceptible core genogroup. Later in 2022, an allele genetically similar to penA-60, mosaic penA-237, was identified in the UK (H22-722) and France (F92) with high-level ceftriaxone MICs and both belonged to MLST-1901. In this study, we assessed phylogenomic relatedness and antimicrobial resistance (AMR) determinant profiles of these three isolates with high-level ceftriaxone MICs among a global collection of 2,104 genomes belonging to the MLST-1901 core genome cluster group 31, which includes strains separated by a locus threshold of 200 or fewer differences (Ng_cgc_200). Recombination events in and around the penA coding region were catalogued and potential sources of inter species recombinant DNA were also inferred. The global population structure of MLST-1901 core genogroup falls into 4 major lineages. Isolates GCWGS-10723, F92, and H22-722 clustered within Lineage 1, which was dominated by non-mosaic penA-5 alleles. These three isolates formed a clade within Lineage 1 that consisted of isolates from North America and southeast Asia. Neisseria subflava and Neisseria sicca were identified as likely progenitors of two independent recombination events that may have led to the generation of mosaic penA-60 and penA-237, within a possible non-mosaic penA-5 background. Our study suggests that there are multiple evolutionary pathways that could generate concerning mosaic penA alleles via homologous recombination of historically susceptible Ng lineages with Neisseria commensals. Enhanced surveillance of gonococcal strains and Neisseria commensals is crucial for understanding of the evolution of AMR, particularly in less-studied regions (e.g., Asia), where high-level ceftriaxone MICs and multi-drug resistance are more prevalent.

Effects of doxycycline post-exposure prophylaxis for prevention of sexually transmitted infections on gonorrhoea prevalence and antimicrobial resistance among men who have sex with men in the USA: a modelling study

Doxycycline post-exposure prophylaxis (PEP) has been shown to be efficacious for the prevention of bacterial sexually transmitted infections, but resistance implications for Neisseria gonorrhoeae remain unknown. We aimed to use a mathematical model to investigate the anticipated impact of doxycycline PEP on the burden of gonorrhoea and antimicrobial resistance dynamics in men who have sex with men (MSM) in the USA. Using a deterministic compartmental model, characterising gonorrhoea transmission in a US MSM population comprising three sexual activity groups defined by annual partner turnover rates, we introduced doxycycline PEP at various uptake levels (10-90%) among those with high sexual activity. Infections were stratified by symptom status and resistance profile (ie, susceptible, ceftriaxone-resistant, tetracycline-resistant, or dual-resistant), with ceftriaxone the treatment for active infection. As resistance to tetracycline, not doxycycline, is monitored and reported nationally, we used this as a proxy for doxycycline PEP resistance. We compared the 20-year prevalence, incidence rates, and cumulative incidence of gonococcal infection, resistance dynamics (time to 5% prevalence of ceftriaxone resistance, 5% prevalence of dual resistance, and 84% prevalence of tetracycline resistance), and antibiotic consumption with baseline (ie, no doxycycline PEP). Uptake of doxycycline PEP resulted in substantial reductions in the prevalence and incidence of gonorrhoea, but accelerated the spread of tetracycline resistance. The maximum reduction in prevalence over 20years compared with no uptake ranged from 40·3% (IQR 15·3-83·4) with 10% doxycycline PEP uptake to 77·4% (68·4-84·9) with 90%uptake. Similarly, the maximum reduction in the incidence rate ranged from 38·6% (14·1-83·6) with 10% uptake to 77·6% (68·1-84·7) with 90% uptake. Cumulative gonococcal infections were reduced by a median of 14·5% (IQR8·4-21·6) with 10% uptake and up to 46·2% (26·5-59·9) with 90% uptake after 5years, and by 6·5% (3·4-13·0) with 10% uptake and 8·7% (4·3-36·2) with 90% uptake by 20years. In almost all scenarios explored, doxycycline PEP lost clinical effectiveness (defined as 84% prevalence of tetracycline resistance) within the 20-year period, but itslifespan ranged from a median of 12·1years (IQR 9·9-15·7) with 10% uptake to 1·6years (1·3-1·9) with 90%uptake. Doxycycline PEP implementation had minimal impact on extending the clinical lifespan of ceftriaxone monotherapy (5·0years [IQR 4·0-6·2]), with the median time to 5% prevalence of resistance ranging from 4·8years (3·9-6·0) for 90% uptake to 5·0years (4·1-6·2) for 10% uptake. Similarly, the median time to 5%prevalence of dual resistance to ceftriaxone and tetracycline ranged from 4·8years (3·9-6·0) for 90% uptake to 5·8years (4·8-7·4) for 10% uptake. Median decrease in ceftriaxone consumption for high doxycycline PEP uptake levels compared with baseline ranged from 41·7% (27·0-54·3) for 50% uptake to 50·2% (29·3-62·7) for 90% uptake at 5years, but dropped to 11·8% (6·9-32·0) for 50% uptake and 12·1% (7·0-41·6) for 90% uptake after 20years. Notwithstanding the clear benefits of doxycycline PEP for other sexually transmitted infections, for Ngonorrhoeae, model findings suggest that doxycycline PEP is an effective but impermanent solution for reducing infection burden, given eventual selection for resistant strains. This finding presents a challenge for policy makers considering strategies for doxycycline PEP implementation and oversight: the need to balance the clear, short-term clinical benefits with the risk of harm via antimicrobial resistance. US Centers for Disease Control and Prevention, National Institute of Allergy and Infectious Diseases.

Mpox outbreak among men who have sex with men in Kraków, Poland; June 2022-November 2022.

Mpox is a zoonotic disease caused by the mpox virus (MPXV). Sporadic cases reported before 2022 were almost universally linked to direct contact with animals or travel to endemic regions in Africa. However, in 2022, a significant shift occurred with human-to-human transmission, leading to a global outbreak in 117 countries. In Poland, 217 cases of mpox were reported. The aim of this study was to evaluate the epidemiology, clinical presentations, laboratory findings, and co-infections with sexually transmitted pathogens among patients with mpox. A retrospective analysis of all mpox cases diagnosed at the University Hospital in Kraków between June 2022 and November 2022 was performed. Forty-five patients aged 19-48 years (median age 34) were diagnosed with mpox. Most of them were male 44 (98%) and 42 (93%) identified themselves as men who have sex with men (MSM). Twenty-two (49%) of the patients had previously been diagnosed with HIV. During mpox diagnosis, new cases of sexually transmitted infections (STIs) were identified. Specifically, 4 patients (9%) were newly diagnosed with HIV, 5 with syphilis, 4 with Chlamydia trachomatis, 3 with Neisseria gonorrhoeae infection, and 2 with Ureaplasma urealyticum infection; among this group, 4 patients (33%) had more than one sexually transmitted co-infection. The mpox outbreak in Kraków in 2022 primarily affected MSM and was transmitted through sexual contact. Healthcare professionals should be aware of emerging STIs. Educational initiatives should emphasize the importance of vaccinations, safe sexual practices and regular testing.

Prevalence of bacterial vaginosis, sexually transmitted infections and their association with HPV infections in asymptomatic women attending antenatal care in Ethiopia.

Sexually transmitted infections (STIs) and human papillomavirus (HPV) infections are common among women of reproductive age and can lead to infertility, adverse pregnancy outcomes, neonatal infections and cervical cancer. In countries with limited medical coverage, untreated infections contribute to high morbidity. This study aimed to expand the current knowledge on the prevalence of bacterial vaginosis (BV) and STIs in pregnant Ethiopian women and assess the association of these conditions with HPV infections. Socio-demographic data and vaginal lavage samples were collected from 779 asymptomatic women aged 18 to 45 years (median age, 25.9 years) attending antenatal care in seven centres across Ethiopia. Multiplex polymerase chain reaction was used to test for BV, Chlamydia trachomatis, Trichomonas vaginalis, Neisseria gonorrhoeae, herpes simplex virus types 1 and 2 (HSV-1/2), Mycoplasma, Ureaplasma, Candida species and HPV. Overall, 26.8% (95% confidence interval (CI): 23.7-29.9) of women tested positive for BV or one of the following STIs: C. trachomatis, T. vaginalis, N. gonorrhoeae, Mycoplasma genitalium, HSV-1/2 or Ureaplasma urealyticum. Additionally, 22.1% tested positive for at least one high-risk HPV type. Chlamydia trachomatis and HSV-2 were significantly more common among women who were positive for HPV and high-risk HPV. This study reveals a high prevalence of asymptomatic pregnant women who are positive for BV, STIs or HPV, putting them at risk of adverse pregnancy outcomes, secondary infertility or cervical cancer in a country with limited medical coverage. Screening and treating these women could be crucial in reducing morbidity.

Gene Expression Of Some Antibiotic Resistance Mechanisms In Neisseria Gonorrhoeae Isolated From Iraqi Patients

Background: Treatment of N. gonorrhoeae infection is restricted by the rising commonness of multidrug-resistance strains. The point of this study was to evaluate antibiotic resistance and mRNA expression of penA, pilQ, norM, farA, and mtrE of N. gonorrhea. Method: one hindered samples collected from patients infected with N. gonorrhoeae. The bacteria were isolated and diagnosed using culture media and biochemical tests. Using the MIC method, the sensitivity of bacteria to penicillin, ciprofloxacin, ceftriaxone, spectinomycin, tetracycline, and azithromycin was determined. After RNA extraction and cDNA production, gene expression of penA, pilQ, norM, farA, and mtrE was assessed using real time -qPCR. Results: we found all isolates sensitive to ceftriaxone and azithromycin. Clear increase in penA, pilQ, farA, and mtrE gene expression of penicillin-resistant isolates (1.571, 3.135, 0.010 and 34.24 respectively), while norM gene expression decreased in those isolates. High gene expression for pilQ, farA, and mtrE among isolates resistant to ciprofloxacin (0.0041, 0.00072, 0.011 respectively). Tetracycline-resistant isolates appeared to have higher gene expression for penA, norM, farA, and mtrE (0.0033, 8.669, 37.85, 15.84 respectively). Spectinomycin-resistant isolates have high gene expression for penA, pilQ, norM, farA, and mtrE (0.222, 0.021, 0.036, 0.0002, 19.63 respectively) . Moreover, gene expression of penA, norM, and farA increased in males (6.938, 1.095, 7.946 respectively) while the rate of gene expression for pilQ and mtrE increased in females (0.161 and 7.832 respectively). Conclusion: gene expression of penA, pilQ, norM, farA, and mtrE increased in most antibiotics resistance isolates especially of bacterial infection that isolated from males.

Trichomonas vaginalis: comparison of primers for implementation as an in-house PCR in rural Vellore, South India

BackgroundTrichomonas vaginalis (TV) accounts for the highest burden of curable, non-viral sexually transmitted infections worldwide. Prevalence in India ranges from 0.4 to 27.4% in women and 0.0-5.6% in men. In 2015, the prevalence of TV among pregnant women of rural Vellore was 3.11% using Sekisui OSOM® Trichomonas test and culture methods. Molecular methods are the most sensitive, rapid diagnostic tool for Sexually Transmitted Infection’s (STI) albeit cost hinders implementation of commercial platforms. To determine a sensitive, sustainable molecular method, we compared three targets (Adhesin AP65, cytoskeleton Beta-tubulin BTUB 9/2 and TVK 3/7) with the highest published diagnostic accuracy against microscopy, culture and Real Time PCR (RT- PCR).Materials & methodsSix-hundred adult, sexually active women attending the Obstetrics-Gynaecology rural out-patient clinic the Rural Unit for Health and Social Affairs (RUHSA) from July 2020 - February 2021 were enrolled. A vaginal lateral and posterior fornix specimen was inoculated, onsite, into Biomed InPouch® TV culture and smeared onto a slide for fluorescence microscopy using Acridine orange. A flocked nylon swab specimen for PCR was used to determine the sensitivities of the Adhesin AP65, cytoskeleton Beta-tubulin BTUB 9/2 and TVK 3/7 gene targets. Seegene Allplex™ STI Essential Assay, S.Korea was used to confirm TV positives.ResultsNine specimens (9/600, 1.5%) were positive for TV. There was a 100% correlation between Biomed InPouch TV® culture, PCR with TVK 3/7 and RT-PCR while a correlation of 66.6% with BTUB 9/2 and AP65 gene targets. Clinically, 77.7% (n = 7) presented with white-greenish discharge per vagina, 11% (n = 1) with infertility, 22.2% (n = 2) were asymptomatic. Eight of nine patients (88.9%) had co-infections with other bacterial STIs. Prevalence of TV coinfection with Neisseria gonorrhoea was 1.1%.ConclusionCurrent hospital-based prevalence of TV in rural Vellore was 1.5%. Repetitive DNA target TVK 3/7 was more sensitive than AP65 and BTUB 9/2 primers.

Oxydifficidin, a potent Neisseria gonorrhoeae antibiotic due to DedA assisted uptake and ribosomal protein RplL sensitivity.

Gonorrhea, which is caused by Neisseria gonorrhoeae , is the second most reported sexually transmitted infection worldwide. The increasing appearance of isolates that are resistant to approved therapeutics raises the concern that gonorrhea may become untreatable. Here, we serendipitously identified oxydifficidin as a potent N. gonorrhoeae antibiotic through the observation of a Bacillus amyloliquefaciens contaminant in a lawn of N. gonorrhoeae . Oxydifficidin is active against both wild- type and multidrug-resistant N. gonorrhoeae . It's potent activity results from a combination of DedA- assisted uptake into the cytoplasm and the presence of an oxydifficidin-sensitive ribosomal protein L7/L12 (RplL). Our data indicates that oxydifficidin binds to the ribosome at a site that is distinct from other antibiotics and that L7/L12 is uniquely associated with its mode of action. This study opens a potential new avenue for addressing antibiotic resistant gonorrhea and underscores the possibility of identifying overlooked natural products from cultured bacteria, particularly those with activity against previously understudied pathogens.

Antimicrobial activity, toxicity and retrospective clinical effectiveness of Kantinka BA and Kantinka Herbaltics, two multi-component-herbal products used in the management of infectious diseases in Ghana

There is an upsurge in the incidence of persons living with infectious diseases and their associated symptoms. Also, there is increased resistance and high cost of available synthetic antimicrobial therapeutic agents. This calls for screening candidate herbal products to examine the risk-to-benefit ratio for users. Moreover, there are inadequate proven scientific studies to assess the quality, effectiveness, and toxicity of herbal products that traditional medicine practitioners in Ghana commonly use for the management of infectious diseases such as those caused by Neisseria gonorrhoeae, Candida albicans and the symptomatic management of symptoms associated with infections like cough, skin infections among others and gastritis. Kantinka BA and Kantinka Herbaltics, two multicomponent herbal products, have been used to manage the above-mentioned disease conditions. The study aims to evaluate the in vitro antibacterial activity, assess the retrospective clinical effectiveness (clinical responses; the disappearance of presenting signs and symptoms associated with infections, gastritis), and the quality and toxicity of Kantinka BA and Kantinka Herbaltics. The products are registered by the Food and Drugs Authority (FDA). Data on 200 patients who were diagnosed with infectious diseases such as Human Immunodeficiency Virus (HIV) / Acquired Immunodeficiency Syndrome (AIDS) and associated symptoms from January 2018 to June 2018 was obtained from the Adom Herbal Clinic, and the Tafo Government Herbal Medicine Records Unit was assessed. The antibacterial activity of the products was evaluated using the HT-SPOTi method. Phytochemical screening, microbial load, and pH were carried out according to standard procedures. Acute toxicity was carried out according to the Organization for Economic Cooperation and Development (OECD) guideline 425. Phytochemical screening, pH, and microbial load have been established for both products. Binding toxicity studies revealed that the products are non-toxic at a 2000 mg/kg dose. The two products exhibited antimicrobial activities against the test organisms with Minimum Inhibitory Concentration (MIC) and Minimum Lethal Concentration (MLC) determined for Kantinka BA and Kantinka Herbaltics as 5% and 10% and &lt;80, respectively, against C. albicans and N. gonorrhoeae - the signs and symptoms associated with infections disappeared during the management period. The products are safe and may effectively manage some infectious diseases and associated symptoms.

Role of doxycycline post exposure prophylaxis in the prevention of sexually transmitted infections: a narrative review

Sexually transmitted infections (STIs) spread through sexual intercourse. They are caused by pathogens like Neisseria gonorrhoeae, Treponema pallidum, Chlamydia trachomatis, and Mycoplasma genitalium, and can be transmitted through direct contact or contact with genital secretions. Doxycycline is a second-generation tetracycline antibiotic with a broad antimicrobial spectrum, acting on intracellular and extracellular organisms. This review aims to explore the efficacy of doxycycline post-exposure prophylaxis (PEP) in the prevention and treatment of STIs. Some of the areas of uncertainty with the use of doxycycline PEP for STIs include the concern for developing antimicrobial resistance (AMR), the effect on the normal microbiome of the body, public acceptability, drug formulation and dosage and compliance issues. Recent clinical and observational studies have looked at using doxycycline PEP to reduce the transmission of STIs. This review aims to explore these studies to provide a comprehensive account of the current state of doxycycline PEP and its future prospects. We also look at the limitations and areas of improvement in using doxycycline PEP, opening avenues for future research. High-quality studies have shown that using doxycycline PEP can effectively reduce the transmission of STIs in a number of clinical trials as well as systematic reviews., making it a potential standard treatment for high-risk groups such as immunocompromised patients, individuals living with HIV, sex workers, and men who have sex with men (MSM).

Advancing microbial diagnostics: a universal phylogeny guided computational algorithm to find unique sequences for precise microorganism detection.

Sequences derived from organisms sharing common evolutionary origins exhibit similarity, while unique sequences, absent in related organisms, act as good diagnostic marker candidates. However, the approach focused on identifying dissimilar regions among closely-related organisms poses challenges as it requires complex multiple sequence alignments, making computation and parsing difficult. To address this, we have developed a biologically inspired universal NAUniSeq algorithm to find the unique sequences for microorganism diagnosis by traveling through the phylogeny of life. Mapping through a phylogenetic tree ensures a low number of cross-contamination and false positives. We have downloaded complete taxonomy data from Taxadb database and sequence data from National Center for Biotechnology Information Reference Sequence Database (NCBI-Refseq) and, with the help of NetworkX, created a phylogenetic tree. Sequences were assigned over the graph nodes, k-mers were created for target and non-target nodes and search was performed over the graph using the depth first search algorithm. In a memory efficient alternative NoSQL approach, we created a collection of Refseq sequences in MongoDB database using tax-id and path of FASTA files. We queried the MongoDB collection for the target and non-target sequences. In both the approaches, we used an alignment free sliding window k-mer-based procedure that quickly compares k-mers of target and non-target sequences and returns unique sequences that are not present in the non-target. We have validated our algorithm with target nodes Mycobacterium tuberculosis, Neisseria gonorrhoeae, and Monkeypox and generated unique sequences. This universal algorithm is a powerful tool for generating diagnostic sequences, enabling the accurate identification of microbial strains with high phylogenetic precision.