Gonorrhea Research Articles

Gonokokkenartritis bij een 69-jarige man

Gonococcal arthritis in a 69-year-old male A 69-year-old man presented with fever and a painful and swollen left wrist and right ankle joint. Aspiration of the left wrist showed purulent fluid and culture was positive for Neisseria gonorrhoeae. The diagnosis of gonococcal arthritis, a manifestation of disseminated gonococcal infection, was made. History revealed he had unprotected sexual contacts with men. Treatment with intravenous ceftriaxon for 7 days and surgical lavage of the joints led to clinical improvement. Gonorrhea has an increasing incidence in Belgium and is now the most diagnosed sexually transmitted disease (STDs) in men. If untreated, infection with N. gonorrhoeae can lead to a disseminated gonococcal infection, sometimes resulting in serious complications such as arthritis, osteomyelitis, and meningitis. A thorough sexual history is essential in cases of acute monoarthritis or oligoarthritis. Joint aspiration with gram stain, culture and/or polymerase chain reaction (PCR) is diagnostic. Treatment of disseminated gonococcal infection consists of intravenous ceftriaxon. Surgical drainage may be necessary in case of purulent arthritis. Notification and testing of sexual partners is crucial to prevent further spread of N. gonorrhoeae. This case illustrates the importance of considering N. gonorrhoeae as a cause of septic arthritis in sexually active individuals. Early recognition and adequate treatment can prevent complications and further transmission of infections. Physicians should be alert to the increasing incidence of gonorrhoea and its potential complications.

Doxycycline Postexposure Prophylaxis and Bacterial Sexually Transmitted Infections Among Individuals Using HIV Preexposure Prophylaxis

Doxycycline postexposure prophylaxis (doxyPEP) has been shown to decrease the incidence of bacterial sexually transmitted infections (STIs) among people assigned male sex at birth in clinical trials, but data from clinical practice are limited. To describe early uptake of doxyPEP and evaluate changes in STI incidence following doxyPEP initiation. This retrospective cohort study of adults (aged ≥18 years) dispensed HIV preexposure prophylaxis (PrEP) at Kaiser Permanente Northern California during November 1, 2022, to December 31, 2023, examined electronic health record data to compare HIV PrEP users dispensed and not dispensed doxyPEP and rates of bacterial STIs before and after starting doxyPEP. Individuals were followed up from their first recorded STI test on or after November 1, 2020, until December 31, 2023, or discontinuation of health plan membership. Pharmacy dispensing data were used to define doxyPEP recipients. Demographic and clinical characteristics were compared between individuals dispensed and not dispensed doxyPEP. Primary outcomes were incident chlamydia, gonorrhea, or infectious syphilis measured as quarterly STI positivity (proportion of individuals testing positive at least once per quarter). Among doxyPEP recipients, rate ratios (RRs) compared mean quarterly STI positivity from 24 months before to 12 months after starting doxyPEP. In an exploratory analysis, STI trends were evaluated for the full cohort, stratified by receipt of doxyPEP. Among 11 551 HIV PrEP users (mean [SD] age, 39.9 [12.1] years; 95.1% male), 2253 (19.5%) were dispensed doxyPEP, of whom 2228 (98.9%) were male and 1096 (48.6%) had an STI in the year before starting doxyPEP. Compared with individuals not dispensed doxyPEP, doxyPEP recipients were older (mean [SD] age, 40.4 [10.8] vs 39.8 [12.4] years; P = .04) and had used HIV PrEP longer (mean [SD], 4.2 [2.8] vs 3.4 [2.6] years; P < .001), and a higher proportion were commercially insured (2091 [92.8%] vs 8270 [88.9%]; P < .001). Among doxyPEP recipients, quarterly chlamydia positivity decreased from 9.6% (95% CI, 9.0%-10.3%) before starting doxyPEP to 2.0% (95% CI, 1.5%-2.6%) after starting doxyPEP (RR, 0.21; 95% CI, 0.16-0.27; P < .001), with significant declines for each anatomic site of infection. Quarterly gonorrhea positivity decreased from 10.2% (95% CI, 9.6%-10.9%) before starting doxyPEP to 9.0% (95% CI, 8.0%-10.1%) after starting doxyPEP (RR, 0.88; 95% CI, 0.77-1.00; P = .048); site-specific declines were significant for rectal (RR, 0.81; 95% CI, 0.67-0.97; P = .02) and urethral (RR, 0.56; 95% CI, 0.40-0.79; P = .001) gonorrhea, but not pharyngeal gonorrhea. Quarterly syphilis positivity decreased from 1.7% (95% CI, 1.4%-1.9%) before starting doxyPEP to 0.3% (95% CI, 0.2%-0.6%) after starting doxyPEP (RR, 0.20; 95% CI, 0.11-0.37; P < .001). Positivity for STIs remained stable in individuals not dispensed doxyPEP. This study found that receipt of doxyPEP was associated with substantial declines in chlamydia and syphilis incidence and modest declines in urethral and rectal gonorrhea incidence among individuals using HIV PrEP. These findings suggest that doxyPEP may offer substantial benefits for reducing population-level STI transmission with broader implementation.

The Impacts and Consequences of Sexually Transmitted Infections in the United States.

Millions of people in the United States are affected by sexually transmitted infections (STIs) every year, with profound consequences for the individual, their community, and society at large. In this review, we aim to summarize the epidemiology of four STIs: C. trachomatis, N. gonorrhoeae, T. pallidum, and HSV, and to highlight the consequences of those infections among individuals and the healthcare system. Untreated N. gonorrhoeae, C. trachomatis, T. pallidum, or HSV infection can result in female infertility, stillbirth, premature birth, and low birth weight. As many as 10% of incident HIV infections among men who have sex with men in the United States have been attributed to either N. gonorrhoeae or C. trachomatis infection. In one year in the United States, incident C. trachomatis infection resulted in $824 million lifetime medical costs and a loss of 1,541 lifetime quality-adjusted life years (QALYs) among men and 111,872 among women, while incident N. gonorrhoeae infection led to $323 million in lifetime medical costs, and a loss of 386 QALYs among women and 12,112 among men. Incident T. pallidum infection in one year resulted in $206 million in medical costs and a loss of 13,349 QALYs among both men and women, while genital herpes led to $107 million in medical costs and a loss of 33,100 QALYs. STI-attributable infertility alone resulted in more than $190 million in direct medical costs. Cumulatively, STIs lead to substantial financial costs to individuals and the health system, as well as long-term reductions in quality of life.

New genotypes in the structure of the Russian population of Neisseria gonorrhoeae

Currently, despite the persistently high incidence of gonococcal infection, the rapid development of Neisseria gonorrhoeae resistance to antimicrobial agents increases the risk of untreatable forms of gonorrhea. Monitoring the spread of antimicrobial-resistant N. gonorrhoeae is a major priority for national surveillance programs. Started in 2021-2022 in Russian Federation, it showed that the trend of the increased sensitivity to penicillins, tetracyclines, fluoroquinolones, and macrolides was replaced by an increase in the number of strains resistant to these drugs, including multi-resistant ones. Molecular typing of strains revealed changes in the genetic structure of the N. gonorrhoeae population, which was long considered to be very conservative; in particular, a significant increase in strains of the new MLST genogroups, previously rarely seen, was identified. Whole-genome sequencing data showed the formation of a separate clusters consisting of strains within the MLST10314, MLST9363 and MLST8143 genogroups. A large number of single nucleotide polymorphisms in the genes of the mtrCDE and mtrR efflux system were revealed. The obtained results indicate the need to enhance both measures to control the spread of antibiotic-resistant N. gonorrhoeae strains and continue whole-genome sequencing of the Russian N. gonorrhoeae population to identify strains with genetic determinants of antimicrobial resistance.

Minimum inhibitory concentrations of extended spectrum cephalosporins: A systematic review and meta-analysis of Neisseria gonorrhoeae treatment failures.

Neisseria gonorrhoeae is one of the recognised global antimicrobial resistance priorities. Extended spectrum cephalosporins, the last remaining reliable antimicrobial, increasingly fail to clear N. gonorrhoeae infections, especially pharyngeal gonorrhoea, leading to limited future treatment options. We conducted a systematic review and meta-analysis of gonococcal treatment failures and compared the minimum inhibitory concentrations (MIC) of isolates from pharyngeal and extra-pharyngeal anatomical sites (PROSPERO registration: CRD42020189101). The overall pooled mean MIC for cefixime was 0.17 mg/L (95% [CI]: 0.07, 0.41) and ceftriaxone was 0.10 mg/L (95% [CI]: 0.05, 0.22). For cefixime, the mean MIC estimates for pharyngeal and extra-pharyngeal treatment failures were 0.05 mg/L (95% [CI]: 0.02, 0.14) and 0.29 mg/L (95% [CI]: 0.11, 0.81), and for ceftriaxone 0.09 mg/L (95% [CI]: 0.03, 0.22) and 0.14 mg/L (95% [CI]: 0.03, 0.73), respectively. The pooled mean MICs for pharyngeal isolates are below the phenotypic European Committee on Antimicrobial Susceptibility Testing (EUCAST) resistance breakpoint for both antimicrobials (>0.125 mg/L). Our findings underscore the need to review the current resistance breakpoints used for pharyngeal infection and the urgency to establish international standards for MIC testing, and advance efforts of the World Health Organization's global action plan to control the spread and impact of antimicrobial resistance in N. gonorrhoeae. Ongoing susceptibility testing of gonococcal isolates and surveillance of treatment failures are central to informing appropriate public health responses.

The Effect of Using a Standardized Questionnaire on Sexual History Documentation and Testing to Diagnose Gonorrhea and Chlamydia Among Men Who have Sex with Men with HIV.

Background: Most Neisseria gonorrhoea (GC) and Chlamydia trachomatis (CT) infections in men who have sex with men (MSM) are diagnosed at extragenital sites. However, testing at these sites is often lacking. The purpose of this study was to determine if a standardized questionnaire administered by physicians and clinical assistants improves documentation of sex activity and increases extragenital testing and diagnoses of GC and CT among MSM.Methods: A standardized sexual history questionnaire was implemented on 11/1/2022. Electronic medical records of 664 MSM with HIV, including 1064 encounters, were reviewed to compare pre- and post- implementation sexual history documentation, adequacy of documentation, extragenital GC and CT testing, and GC and CT diagnoses. Analysis included Chi-square and exact tests and logistic regression adjusting for physician cluster effects.Results: The standardized questionnaire was used by 53.7% of physicians and 85.9% of coordinators. Documentation of whether sexual activity occurred increased from 79.3% [95%confidence interval (CI) 0.758- 0.828] in the pre-intervention pre-COVID-19 period to 95.2% (95% CI 0.925-0.970) in the post-intervention period with an adjusted odds ratio of 4.7 (95% CI 2.7-8.8). Specific questions about anal and oral sex increased from 42.0% to 88.1% (p < 0.001) and 23.7% to 88.7% (p < 0.001), respectively. Anal and pharyngeal testing increased from 14.4% to 20.2% (p = 0.040) and 17.2% to 23.3% (p = 0.045), respectively.Conclusions: This study demonstrates that using a standardized questionnaire during clinical encounters can improve documentation of sexual activity and testing for GC and CT at extragenital sites.

Ceftriaxone-and-cefixime-resistant Neisseria gonorrhoeae in a 26-year-old-male with gonococcal urethritis: a case report

Ceftriaxone-and-cefixime-resistant Neisseria gonorrhoeae in a 26-year-old-male with gonococcal urethritis: a case report

Long-term 4CMenB vaccine effectiveness against gonococcal infection at four years post program implementation: observational case-control study

Long-term 4CMenB vaccine effectiveness against gonococcal infection at four years post program implementation: observational case-control study

Helichrysum populifolium Compounds Inhibit MtrCDE Efflux Pump Transport Protein for the Potential Management of Gonorrhoea Infection.

The progressive development of resistance in Neisseria gonorrhoeae to almost all available antibiotics has made it crucial to develop novel approaches to tackling multi-drug resistance (MDR). One of the primary causes of antibiotic resistance is the over-expression of the MtrCDE efflux pump protein, making this protein a vital target for fighting against antimicrobial resistance (AMR) in N. gonorrhoeae. This study was aimed at evaluating the potential MtrCDE efflux pump inhibitors (EPIs) and their stability in treating gonorrhoea infection. This is significant because finding novel EPIs would allow for the longer maintenance of antibiotics at therapeutic levels, thereby prolonging the susceptibility of currently available antibiotics. A virtual screening of the selected Helichrysum populifolium compounds (4,5-dicaffeoylquinic acid, apigeninin-7-glucoside, and carvacrol) was conducted to evaluate their potential EPI activity. An integrated computational framework consisting of molecular docking (MD), molecular mechanics generalized born, and surface area solvation (MMGBSA) analysis, molecular dynamics simulations (MDS), and absorption, distribution, metabolism, and excretion (ADME) properties calculations were conducted. Of the tested compounds, 4,5-dicaffeoylquinic acid revealed the highest molecular docking binding energies (-8.8 kcal/mol), equivalent MMGBSA binding free energy (-54.82 kcal/mol), indicative of consistent binding affinity with the MtrD protein, reduced deviations and flexibility (root mean square deviation (RMSD) of 5.65 Å) and, given by root mean square fluctuation (RMSF) of 1.877 Å. Carvacrol revealed a docking score of -6.0 kcal/mol and a MMGBSA computed BFE of -16.69 kcal/mol, demonstrating the lowest binding affinity to the MtrD efflux pump compared to the remaining test compounds. However, the average RMSD (4.45 Å) and RMSF (1.638 Å) of carvacrol-bound MtrD protein showed no significant difference from the unbound MtrD protein, except for the reference compounds, implying consistent MtrD conformation throughout simulations and indicates a desirable feature during drug design. Additionally, carvacrol obeyed the Lipinski rule of five which confirmed the compound's drug-likeness properties making it the most promising EPI candidate based on its combined attributes of a reasonable binding affinity, sustained stability during MDS, its obedience to the Lipinski rule of five and compliance with drug-likeness criteria. An in vitro validation of the potential EPI activities of H. populifolium compounds confirmed that 4,5-dicaffeoylquinic acid reduced the expulsion of the bis-benzimide dye by MtrCDE pump, while carvacrol showed low accumulation compared to other compounds. While 4,5-dicaffeoylquinic acid demonstrated the highest binding affinity in computational analysis and an EPI activity in vitro, it showed lower stability compared to the other compounds, as indicated in MDS. This leaves carvacrol, as a better EPI candidate for the management of gonorrhoea infection.

Increase in Neisseria gonorrhoeae infections after ending COVID-19 lockdown measures in Amsterdam, the Netherlands

ObjectiveVarious European countries noted increasing Neisseria gonorrhoeae (Ng) positivity among cis-gender women and heterosexual men. We assessed changes over time in Ng among men who exclusively have sex with men...

A disseminated gonococcal infection with spinal locations.

A disseminated gonococcal infection with spinal locations.

Multicenter Clinical Performance Evaluation of the NeuMoDx™ CT/NG Assay 2.0.

Given the continued increases in rates of both Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) infection, additional diagnostic assays may be useful in increasing access to testing for these sexually transmitted infections. We evaluated the performance of the NeuMoDx™ CT/NG Assay 2.0 on the NeuMoDx-96 and NeuMoDx-288 Molecular Systems. The clinical sensitivity and specificity of the assay was assessed when used with: 1) endocervical swabs; 2) self-, and clinician-collected vaginal swabs; and 3) first-catch urine specimens (female and male). Results were compared to a patient infection status based on US Food and Drug Administration (FDA)-cleared assays. The NeuMoDx CT/NG Assay 2.0 demonstrated high sensitivity and specificity in both symptomatic and asymptomatic participants. All specimen types other than endocervical swabs had >95% sensitivity and > 99% specificity for both pathogens. For endocervical samples, sensitivity was 93.2% and 93.3% for CT and NG, respectively. There was no difference in performance based on platform. The frequency of invalid results was low (<1%). The NeuMoDx CT/NG Assay 2.0 demonstrated performance similar to currently FDA-cleared assays, with the added choice of a moderate- (96-sample) or a high-throughput (288-sample) platform. The system therefore offers solutions to laboratories running lower volumes of testing that may obviate the need for outsourcing to larger reference laboratories.

Impact of Varying Pre-exposure Prophylaxis Programs on HIV and Neisseria gonorrhoeae Transmission Among MSM in the Netherlands: A Modeling Study.

In 2019, a 5-year pre-exposure prophylaxis (PrEP) program started in the Netherlands, in which up to 8500 men who have sex with men (MSM) can obtain PrEP and 3-monthly consultations with HIV/STI testing. We assessed the impact of the PrEP program on transmission of HIV and Neisseria gonorrhea (NG) among MSM in the Netherlands and examined prospective variations of the program after 2024. We used an agent-based model to estimate the effect of the PrEP program. For hypothetical prospective PrEP programs starting in 2024, we varied the capacity (8,500; 12,000; 16,000 participants) and consultation frequency (3-monthly; 6-monthly; 70% 3-monthly and 30% 6-monthly). At a capacity of 8,500 participants and 3-monthly consultations, the PrEP program could lead to 3,140 [95% credible interval (95% CrI): 1,780-4,780] and 27,930 (95% CrI: 14,560-46,280) averted HIV and NG infections, requiring 316,050 (95% CrI: 314,120-317,580) consultations. At a capacity of 16,000 participants, the programs with 3-monthly consultations and 6-monthly consultations could lead to comparable number of averted HIV [3,940 (95% CrI: 2,420-5,460), and 3,900 (2,320-5,630) respectively] and NG infections [29,970 (95% CrI: 15,490-50,350), and 29,960 (95% CrI: 13,610-50,620) respectively], while requiring substantially different number of consultations: 589,330 (95% CrI: 586,240-591,160) and 272,590 (95% CrI: 271,770-273,290), respectively. Continuation of a PrEP program could lead to a substantial reduction in HIV and NG transmission. More infections could be averted if the number of participants is increased. In turn, the consultation frequency could be reduced without reducing the number of averted infections if capacity is increased.

Protection against N. gonorrhoeae induced by OMV-based Meningococcal Vaccines are associated with cross-species directed humoral and cellular immune responses.

Limited protective immunologic responses to natural N. gonorrhoeae infection and a lack of knowledge about mechanisms of protection have hampered development of an effective vaccine. Recent studies in humans and mice have found meningococcal outer membrane vesicle-containing vaccines (OMV) induce cross species immune responses against gonococci and are associated with protection. The exact mechanisms or how humoral and cellular immunity are related to protection, remain unclear. To study this, we immunized mice with two meningococcal OMV-containing vaccines known to accelerate clearance of N. gonorrhoeae , 4CMenB and OMV from an engineered N. meningitidis strain lacking major surface antigens PorA, PorB, and Rmp (MC58 ΔABR). We assessed serologic and cellular immune signatures associated with these immunizations and assessed bacterial clearance in the mice using a vaginal/cervical gonococcal infection model. Mice immunized with 4CMenB or MC58 ΔABR demonstrated shortened courses of recovery of vaginal N. gonorrhoeae compared to control mice immunized with alum alone. Vaccination with 4CMenB or MC58ΔABR OMV elicited serum and vaginal cross-reactive anti-Ng-OMV antibody responses that were augmented after vaginal challenge with N. gonorrhoeae . Further, splenocytes in 4CMenB and MC58 ΔABR immunized mice exhibited elevated cytokine production after restimulation with heterologous N. gonorrhoeae OMV when compared to splenocytes from Alum immunized mice. We further tested for correlations between bacterial burden and the measured anti-gonococcal immune responses within each vaccination group and found different immunologic parameters associated with reduced bacterial burden for each vaccine. Our findings suggest the cross-protection against gonococcal infection induced by different meningococcal OMV vaccines is likely multifactorial and mediated by different humoral and cellular immune responses induced by these two vaccines.

Performance and acceptability of self-collected specimens for diagnosis of rectal and pharyngeal Chlamydia trachomatis and Neisseria gonorrhoeae infections among men who have sex with men in China: a randomized controlled trial

BackgroundExtragenital Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) infections are prevalent among men who have sex with men (MSM). Self-sampling could potentially eliminate barriers to extragenital CT/NG testing for MSM that are hard to reach, who refuse to go for clinician-based testing, or who decline an examination. However, the required evidence to determine whether self-collected specimens are as accurate as clinician-taken specimens in terms of CT/NG diagnostic accuracy was limited in low and middle income countries. We therefore compared self-collected rectal and pharyngeal specimens with clinician-taken specimens for diagnostic accuracy among MSM in China.MethodsThis was a prospective convenience sample from 6 sexually transmitted infection (STI) clinics in China. We randomized the order of self-collected and clinician-taken specimens from the pharynx and rectum, plus first-void urine, for CT/NG detection. Self-sampling performance was compared with clinician-sampling as to agreement, sensitivity, and specificity. The acceptability of self-sampling was evaluated by questionnaire.ResultsAmong the 325 participants, prevalences of rectal CT and NG infections were 13.6% and 5.2% and pharyngeal CT and NG prevalences were 1.5% and 2.8%, respectively. The agreements between the CT tests with the self-collected and clinician-taken specimens were 98.8% (κ = 0.95, 95% CI 0.89-1.00) for rectal site and 99.4% (κ = 0.83, 95% CI 0.60-1.00) for pharyngeal site; and the agreements between NG tests were 99.4% (κ = 0.94, 95% CI 0.86-1.00) for rectal site and 98.2% (κ = 0.72, 95% CI 0.50–0.93) for pharyngeal site. The sensitivity and specificity of self-collected swabs was as follows: rectal CT: 93.0% and 99.6%; pharyngeal CT: 100.0% and 99.4%; rectal NG: 100.0% and 99.4%; pharyngeal NG: 88.9% and 98.4%. Self-collection was highly acceptable, showing that 62.0% MSM preferred self-sampling over clinician-sampling; 90.2% would use self-sampling for detection of CT and NG again.ConclusionsExtragenital screening for CT and NG should be recommended as part of STI services to MSM population. Self-collection of rectal and pharyngeal specimens had good performance for CT and NG tests and acceptability to the target population.Trial registrationThis trial was registered with the Chinese Clinical Trial Registry, ChiCTR2300073473. Registered 12/07/2023.

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

Mpox is a zoonotic disease caused by mpox virus. Sporadic cases reported before 2022 were almost universally linked to a direct contact with animals or travel to endemic regions of Africa. However, in 2022, a significant shift occurred in 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 presentation, laboratory findings, and coinfections with sexually transmitted pathogens among patients with mpox. We retrospectively analyzed all mpox cases diagnosed at the University Hospital in Kraków between June 2022 and November 2022. Forty‑five patients at a median (interquartile range) age of 34 (19-48) years were diagnosed with mpox. Most of them (n = 44; 98%) were men, 42 (93%) identified themselves as men who have sex with men (MSM). Twenty‑two of the patients (49%) had previously been diagnosed with HIV. Together with 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 infection, 3 with Neisseria gonorrhoeae infection, and 2 with Ureaplasma urealyticum infection; in this group, 4 patients (33%) had more than 1 sexually transmitted coinfection. The 2022 mpox outbreak in Kraków primarily affected MSM and was transmitted through sexual contact. Health care professionals should be aware of emerging STIs. Educational initiatives should emphasize the importance of vaccinations, safe sexual practices, and regular testing.

Computational modeling of cyclotides as antimicrobial agents against Neisseria gonorrhoeae PorB porin protein: integration of docking, immune, and molecular dynamics simulations.

Neisseria gonorrhoeae is the bacterium responsible for gonorrhoea, one of the most common sexually transmitted infections (STIs) globally. In 2020, the World Health Organization (WHO) estimated 82.4 million new cases of Neisseria gonorrhoeae infections. Current treatments rely on antibiotics, but the emergence of multi drug resistance (MDR) strains poses a significant threat to public health. This research aims to use computational modeling of cyclotides as antimicrobial agents targeting the Neisseria gonorrhoeae PorB Porin protein to inhibit its pathogenicity. The PorB Porin protein was retrieved from the Protein Data Bank (PDB ID: 4AUI), cleaned, and visualized using Discovery Visual Studio. Physicochemical properties were predicted using ProtParam. Cyclotides were obtained from the CyBase database, with 3D models generated and refined via the Swiss Model for docking studies. HDOCK was used for molecular docking. Toxicity and allergenicity predictions were performed with ToxinPred and AlgPred. A heatmap of the peptide was created using Protein-Sol. Molecular dynamics (MD) simulations were conducted for 100,000 picoseconds using Desmond from Schrödinger LLC, while binding energy was analyzed using MMGBSA. Immune response simulations were done with C-ImmSim 10.1, and peptide simulation in water was performed via WebGro. The protein's GRAVY value is -0.539, indicating moderate hydrophilicity, and its isoelectric point is 9.14, suggesting a fundamental nature. Globa D had the highest docking score (-270.04kcal/mol) and was deemed non-toxic and non-allergenic. MD simulations showed stable protein-ligand interactions, and MMGBSA revealed a low binding energy of -36.737kcal/mol. Immune simulations indicated an effective immune response and peptide simulations demonstrated Globa D's stability in water, making it a potential candidate for pharmaceutical applications. Globa D proved the best drug candidate against Neisseria gonorrhoeae by inhibiting PorB Porin protein chain A. Further in vitro and in vivo studies are recommended to validate these findings and explore clinical applications.

Highly Sensitive Detection of Genomic DNA Using Rapid Real-Time PCR Based on Oscillating Flow

Some individuals are diagnosed with urethritis owing to the detection of gonorrhoea or chlamydia infection through clinical tests; however, there are cases in which neither is detected despite exhibiting symptoms. One of the causative bacteria of non-chlamydial non-gonococcal urethritis is Mycoplasma genitalium, and real-time polymerase chain reaction (qPCR), an accurate examination of causative bacteria for proper drug administration, has been used for its detection. However, given that qPCR is time-consuming, applying it to same-day testing is difficult. Therefore, a rapid qPCR system was developed using a microfluidic device.The microfluidic device used for rapid thermal cycling with oscillating flow in each detection unit was 76 mm × 52 mm × 4 mm in size. The PCR solution was injected into the microchannel from the inlet. The microfluidic device, including the liquid plug, was placed on the chip holder, and three small heaters were embedded into the rapid qPCR system. The temperature of each heater was constantly controlled for reverse transcription (optionally), DNA denaturation and ex-tension by DNA polymerase for PCR. Furthermore, both ends of the microchannel were automatically connected with pneumatic lines using the needles installed in the upper cover of the chip holder when pushed down. This system employed a pair of microblowers as the driving force for the reciprocating liquid transfer. When the microblower was turned off, it quickly released the pressure between the liquid plug and microblower without additional valves, such that the liquid transfer in the microchannel could be stopped at any position immediately. The liquid plug of the PCR solution was reciprocally transferred at programmed timing by switching the two microblowers controlled by the main unit. The detection unit also had an array of three fluorescence detectors aligned along the microchannel. To detect the passage of the PCR solution, the fluorescence of fluorescein mixed with the PCR solution was measured in the middle of the two heaters for denaturation and extension. The fluorescence signal was used to precisely control the stop position on the heater. The remaining two fluorescence detectors were located above the two heaters individually and were used for liquid position detection and evaluation of the amplification of the MgPa gene and internal control.A standard curve was evaluated against gDNA standards of M. genitalium. For the thermal cycle conditions, heating at 96˚C for 10 s, 3 s at 96˚C, and 6 s at 59˚C was repeated 50 times. A dilution series of 0.063 to 5.0×104 copies/run was used as the gDNA concentration of M. genitalium. In addition, to confirm the detection frequency at low concentrations, samples were tested nine times at concentrations where the presence or absence of gDNA in the PCR solution varied statistically. Fig. (a) shows the detection frequency at each concentration of gDNA of M. genitalium. The rapid RT-PCR system was amplified from extremely low concentrations of samples even if the number was < 1 copy/run, although the frequency was not 100%. Fig. (b) shows the standard curve obtained from these amplification curves, which shows good linearity up to 5 copies/run. The PCR efficiency was calculated to be 96.3%. For concentrations of < 5 copies, the success or failure of detection was speculated to depend on the stochastic inclusion of the template DNA in the PCR solution. Fig. (c) shows the detection frequency for each gDNA concentration of M. genitalium and a fitting curve based on the cumulative distribution function (CDF) of the Poisson distribution. The expected value of the maximum likelihood CDF corresponded to 1 copy/run, which would theoretically yield the best LOD. The LOD with a 95% confidence interval (CI) was estimated to be 3 copies/run from this fitting curve. The statistically estimated limit of detection was equivalent to the amplification from a single molecule of genomic DNA, and the PCR efficiency was calculated to be almost 100%. Moreover, the analysis time for a negative judgement obtained without amplification was within 11 min. Thus, the developed rapid RT-PCR system would be useful for the same-day testing of infectious bacteria. Figure 1

The occurrence of single-site pharyngeal Neisseria gonorrhoeae among female sex workers in the Netherlands.

Female sex workers (FSW) have a higher burden of sexually transmitted infections (STI), including Neisseria gonorrhoeae (NG), than the non-sex worker population, mainly due to social vulnerability and work-related factors. This large study evaluated the anatomical site distribution of NG, and specifically the occurrence of single-site pharyngeal NG, among FSW visiting Dutch STI clinics in the Netherlands. Coded STI clinic consultations from FSW attending any Dutch STI clinic between 2016 and 2021, in which tests were performed on all three anatomical sites, according to the Dutch testing policy for FSW, were included in analyses (n = 22,304). To compare the anatomical site distribution of NG among FSW with women in general, 15,494 consultations from women who were tested on three anatomical sites in clinics that performed standard testing at three anatomical sites were used. Descriptive statistics were used to compare the anatomical site distribution of NG and occurrence of single-site pharyngeal NG among FSW and universally tested women. NG was diagnosed among 3.5% (782/22,034) among FSW and 1.7% (271/15,494) among universally tested women (p < 0.001). Pharyngeal NG was diagnosed among 2.3% among FSW, versus 0.8% among universally tested women (p < 0.001). Single-site pharyngeal NG was diagnosed in 19.9% (156/782) of all NG infections, compared to 14.8% (40/271) of NG infections among universally tested women. The substantial proportion pharyngeal NG and single-site pharyngeal NG among FSW, confirms the relevance of pharyngeal testing among this key population for optimal patient management, also in the context of the potential role of pharyngeal NG in antimicrobial resistance.

Prevalence of gonococcal and chlamydial infections among men who have sex with men in sub-Saharan Africa: a systematic review and meta-analysis

BackgroundMen who have sex with men (MSM) are disproportionately affected by sexually transmitted infections (STI) including Neisseria gonorrhoeae (Ng) and Chlamydia trachomatis (Ct). The lack of robust data on STIs among African MSM has limited the development of evidence-based screening strategies. This study aimed at documenting the pooled prevalence of Ng/Ct among MSM in sub-Saharan Africa (SSA).MethodsThis systematic review was performed according to the Preferred Reporting Items for Systematic Review and Meta-analyses (PRISMA) 2020 guidelines. Relevant articles from the following databases were searched: PubMed, Scopus, ISI Web of Science, and the Directory of Open Access Journals (DOAJ). Eligible studies reported on the prevalence of Ng/Ct among the MSM population in SSA. Publication bias was assessed using the Hoy tool, Doi plot, and LFK ratio. Due to heterogeneity among studies, subgroup analyses were performed using the MetaXL add-on tool for Microsoft Excel.ResultsOf 525 articles screened, 20 were selected for inclusion. Six were cross-sectional, four had a prospective cohort study design, and one was an epidemiological study. The pooled prevalence of Ng/Ct in MSM was 27% (95% CI, 19–39%), with an I2 of 98% signifying heterogeneity among the studies. Subgroup analysis by country revealed South Africa had the highest prevalence (38%).DiscussionInterpretationThe high prevalence of Ng/Ct infection among MSM in SSA is of concern.LimitationsDue to limited data available on Ng/Ct prevalence, the true prevalence of SSA and its associated risk factors is uncertain.ConclusionAs the first study to systematically review the available literature on STI prevalence among the MSM population in SSA, it showed the burden of Ng/Ct is higher than in other regions, warranting the strengthening of health systems to improve education, testing, and treatment in MSM population.Systematic review registrationPROSPERO CRD42022327095.