Haycocknema perplexum is a rare and emerging cause of parasitic myositis. Detection and surveillance are of growing importance given the increasing degree of climate conditions conducive to transmission. In this study, we developed a real-time polymerase chain reaction (PCR) method for direct detection of two genomic regions of H. perplexum. The real-time PCR assays, SSU-PCR and COX-1-PCR, targeted the small subunit of nuclear ribosomal RNA and cytochrome oxidase-1 genomic regions, respectively. The performance of the assays was assessed using a panel of H. perplexum samples, both fresh frozen and formalin-fixed paraffin-embedded (FFPE) tissue (n=22, derived from eight patients) and tissue biopsy specimens not derived from H. perplexum (n=8). Both H. perplexum assays showed 92% and 84% sensitivity for SSU and COX-1 targets, respectively, 100% specificity, and negative and positive predictive values of 100% and 93%, respectively. The results indicate that the limit of detection was 10-5 dilution [cycle threshold (Ct) value 40.2] for COX-1-PCR, and 10-3 dilution (Ct value 39.6) for SSU-PCR, making the latter a more sensitive assay for detecting lower concentrations of organism in the patient biopsy. The sensitivity of fresh frozen samples was superior to FFPE samples. All but one sample was negative following treatment. Feasibility of real-time PCR detection of H. perplexum directly from tissue biopsies has been demonstrated for diagnosis, possible test of cure, and could enhance transmission surveillance.

Background:Emerging evidence suggests the 4CMenB vaccine provides moderate protection against gonorrhea, likely due to the close genetic relationship between Neisseria gonorrhoeae and Neisseria meningitidis. 4CMenB has an impact on unencapsulated meningococcal oropharyngeal carriage, where outer membrane proteins are exposed, potentially exerting selective pressure on these strains. We assessed the impact of 4CMenB immunization on gonorrhea disease and oropharyngeal meningococcal carriage among adolescents in the Northern Territory (NT), Australia. Methods:All adolescents aged 14-19 years in the NT were eligible to receive two doses of 4CMenB between 2021 and 2023. Participants had oropharyngeal swabs collected at baseline and 12 months. Vaccine effectiveness (VE) against gonorrhea was assessed using linked notification and immunization data. A Cox proportional hazards model stratified by sex, age, and geography estimated VE. Mixed-effects logistic regression estimated postvaccination odds of meningococcal carriage. Results:A total of 30 650 adolescents were included in the VE analysis, with 9.6% receiving 2 doses of 4CMenB, and 42.4% of the population identifying as Aboriginal or Torres Strait Islanders. Gonococcal notifications were reduced in vaccinated adolescents (VE 38.4%, 95% CI: 18.6-53.4). Overall, meningococcal carriage increased from 4.0% to 6.2% (OR 1.68, 95% CI: 1.14-2.48), driven by genogroup B and nongroupable strains. Carriage of disease-associated meningococci remained stable. Conclusions:4CMenB confers modest protection against gonorrhea in a real-world setting with higher adolescent disease prevalence. These findings are important for women, considering infections are often asymptomatic and have the potential for serious complications. The NT has now introduced a 4CMenB infant and adolescent program.

BACKGROUND:A sustained outbreak of infectious syphilis has been documented in Australia since 2011. Although treatment of syphilis with antibiotics is highly effective, the ongoing spread of this pathogen in Australia suggests that diagnosis may be suboptimal. Molecular detection of syphilis is now commonly performed; however, in many instances clinicians request separate tests for specific pathogens, which may lead to issues in failing to request testing for Treponema pallidum subspecies pallidum, the causative agent of syphilis. METHODS:In this study, we performed routine screening of 5052 samples submitted to our local pathology provider for routine herpes simplex virus investigations. Samples were tested for the presence of T. pallidum. RESULTS:We identified a total of 41 samples (0.8%) which were T. pallidum positive. Of these, the majority of samples reassuringly had requests for T. pallidum testing requested; however, samples from three patients (including a pregnant female) were identified as part of this exercise, who would have otherwise remained undetected and untreated. CONCLUSION:This study demonstrates the clear value in routine screening exercises such as this to improve the timely detection and management of syphilis in Australia.

OBJECTIVES:In remote Australian First Nations communities, the burden of curable sexually transmitted infections (STIs) is highest for young women and men aged 16-29 years and for women is associated with two-fold higher rates of hospitalisations for pelvic inflammatory disease (PID) than for non-First Nations women. Following a randomised trial, decentralised community-led molecular point-of-care (POC) testing for STIs has operated in remote primary care across Australia for more than 7 years, improving uptake and timeliness of treatment for chlamydia, gonorrhoea and trichomonas infections. However, cost-effectiveness remains unknown. METHODS:A decision analytic model was devised to compare costs and outcomes associated with a POC testing programme for chlamydia, gonorrhoea and trichomonas infections in women and men aged 16-29 years seeking care, compared with standard care (laboratory-based testing). The analysis used a government payer perspective and 10-year time horizon. The primary outcome was the cost ($A) per quality-adjusted life year (QALY) gained. Sensitivity analyses examined uncertainty around the results. RESULTS:Based on a combined testing positivity rate of 36% and 29% for chlamydia, gonorrhoea and trichomonas for women and men, respectively, the POC testing programme, compared with laboratory testing, produced an estimated incremental cost per QALY ratio (ICER) of $A19 714 (95% CIs $A19 608 to $A19 821) over 10 years. Among those with an STI, the POC testing programme was predicted to reduce diagnosed PID by 30% and preterm/low birth weight babies by 17%. Sensitivity analyses indicated that the ICER was most sensitive to the probability of infection and receiving treatment within 2 days, based on a willingness-to-pay threshold of $A50 000. CONCLUSION:This health economic evaluation indicates that a scaled molecular POC testing programme for the management of STIs in remote primary care settings is cost-effective compared with standard care. Sustained POC testing in this setting is likely to improve reproductive health outcomes.

BackgroundSexually transmitted infections (STIs) are among the most common infectious diseases globally. In the Asia Pacific (APAC) region, syndromic management is widely practiced, though approaches to aetiological testing vary. Thirteen experts from across APAC convened to develop evidence-based recommendations for testing the four most common infections associated with STI syndromes: Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, and Mycoplasma genitalium.Methods and ResultsThe consensus was developed according to a modified Delphi-based approach. Thirteen experts of multidisciplinary background, representing nine geographical areas in APAC, contributed to the discussion. Twenty-two statements were developed addressing five clinical questions: (1) testing and screening recommendations for these four STIs and frequency of screening, where possible; (2) antimicrobial resistance testing; (3) optimal specimen types for testing; (4) role of self-/patient-collected samples for testing; and (5) role of sample pooling (multiple sites from an individual or pooling of the same sample type from different individuals) in testing. All statements achieved strong consensus, with ≥90% agreement among the experts.ConclusionThese recommendations, while not replacing the existing guidelines, will provide evidence-based pragmatic approach on the testing of four common STIs, which can be adapted for specific settings, depending on available resources.

BACKGROUND:Increasing antimicrobial resistance to ceftriaxone in Neisseria gonorrhoeae presents a public health crisis. We aimed to identify a target associated with ceftriaxone resistance to support the development of rapid diagnostic tests. METHODS:In this observational study, we included N gonorrhoeae isolates with linked penA (NEIS1753) locus and ceftriaxone minimum inhibitory concentration (MIC) data. We compiled and collated isolate-level data from the international public databases PubMLST and Pathogenwatch, and publications up until March 1, 2025. We determined ceftriaxone resistance using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoint of an MIC of 0·250 mg/L or more and the WHO alert value of an MIC of 0·125 mg/L or more. We examined ceftriaxone resistance dynamics, with the sensitivity for susceptibility and resistance based on the constructed penA genotypes calculated. FINDINGS:We compiled 18 689 unique N gonorrhoeae isolates from 71 countries across all WHO regions, collected between 1971 and 2024. Using the EUCAST breakpoint, the overall ceftriaxone resistance rate was 2·4%, with evidence of increasing resistance over time. Analysis of MIC distributions revealed that two alleles, penA 60.001 and penA 237.001, both of the VMTAKGGP genotype, had MIC distributions predominantly above the resistance breakpoint. The VMTAKGGP genotype, defined by the A311V mutation, was associated with a sensitivity of 83·2% (95% CI 79·4-86·6) for detecting ceftriaxone resistance and 99·8% (99·7-99·9) for detecting sensitivity. INTERPRETATION:The A311V mutation is a key marker of ceftriaxone resistance. This target could inform the development of a molecular assay to detect ceftriaxone resistance. FUNDING:Australian Research Council.

Molecular typing can identify relationships between M. genitalium strains and antimicrobial resistance and demographic data. We examined the association of mgpB sequence types (STs) with sex/sexual orientation, antimicrobial resistance and geographical location for M. genitalium in Australia. Sequence data derived from previous studies in Victoria and Queensland were obtained from 170 M. genitalium samples for the mgpB, 23 S rRNA, and parC genes. An additional 55 M. genitalium samples from Victoria were sequenced for the same three genes in this study. A combined data set of 225 samples collected between 2017 and 2019 were examined for associations between mgpB ST and (i) sex/sexual orientation, (ii) macrolide and fluoroquinolone resistance, and (iii) geographical location using chi-square test. Overall, 66 mgpB STs were identified; the most common were ST-7 (17.9

Since the introduction of Neisseria gonorrhoeae nucleic acid amplification tests (NG-NAATs) into routine clinical use, false-positive results caused by cross-reaction with non-gonococcal Neisseria species have been an issue, particularly in specimens from the pharynx. Therefore, since 2005 in Australia, a confirmatory assay has been recommended, with a positive result issued only when both assays are concordant. At the request of the Public Health Laboratory Network (PHLN) Australia, the National Neisseria Network (NNN) met to review the 2015 PHLN NG-NAATs Guidelines in October 2024, in the context that some later generation N. gonorrhoeae NAATs have claims for testing pharyngeal samples without the need for supplemental testing for confirmation. Adequacy of performance in this context was considered by the NNN as a positive predictive value of 95% in line with World Health Organization guidance. Based on the 2024 review, it is recommended that: Supplementary testing continue to be performed for all non-urogenital (pharyngeal and rectal) samples. Supplementary testing be at the discretion of individual laboratories, based on local validation data demonstrating adequate performance based on WHO recommendations, for urogenital samples. Additional testing should continue to be considered when testing low-risk populations.

Antimicrobial resistance (AMR) associated with the leading sexually transmitted infections (STIs) such as gonorrhoea, chlamydia, and trichomoniasis has resulted in significant challenges in the prevention and treatment of these infections using current approaches. To address this, the intravaginal sol-gel platform holds immense potential, as the female reproductive tract (FRT) is the primary site of invasion and colonisation of STI-causing organisms, herein, Neisseria gonorrhoeae. The lead sol-gels, F5 and F9 demonstrate relatively low doxycycline hyclate (DOXH) tissue permeability (10.18 ± 1.56 % and 4.49 ± 1.53 %, respectively), while displaying substantive tissue deposition (2372.95 ± 135.79 µg/g and 2187.73 ± 95.29 µg/g respectively) at 8 h in ex vivo bovine vaginal mucosal tissue. Furthermore, the attenuation of the deleterious effect of DOXH by the sol-gel platform on HeLa cell viability presents it as a safe drug delivery vehicle. The distinct possibility of dose reduction is demonstrated by the negligible differences in the zone of inhibition (ZoI) for sensitive isolates of N. gonorrhoeae with half-strength (0.25 % w/w) and full-strength (0.5 % w/w) DOXH sol-gels. In summary, intravaginal delivery using engineered DOXH-infused sol-gels presents a demonstrable solution to STI prevention/treatment with the potential to reduce AMR risk through localised drug delivery, dose reduction, and increased patient compliance.

AimTo explore the oral microbiota in healthy individuals with and without oral human papillomavirus (HPV) infection and identify any features or changes in the oral microbiota associated with intermediate HPV infection.Materials and methodsPacBio HiFi sequencing of the whole 16S rRNA gene was performed on 128 saliva samples from a previously characterized cohort (64 HPV-positive and 64 HPV-negative). Statistical analysis of alpha- and beta-diversity, as well as taxonomic composition (differential abundance testing), were used to determine differences between-subject groups.ResultsWe demonstrated (1) significant differences in the abundance of Streptococcus salivarius in oral HPV-negative males; (2) males with low-risk HPV had an increased abundance of several species and (3) decreased abundance of Streptococcus salivarius and Streptococcus parasanguinis. For women, (4) oral microbiota Shannon diversity was significantly lower in HPV-positive subjects compared to HPV-negative subjects and (5) oral community structure (beta-diversity) was significantly different when stratified by HPV status.ConclusionsIntermediate oral HPV infection is associated with several perturbations in the abundance of some bacterial taxa in the oral microbiota, which differ between males and females. Further research is required to determine whether these changes contribute to oral carcinogenesis.

Haycocknema perplexum is a rare and emerging cause of parasitic myositis. Detection and surveillance are of growing importance given the increasing degree of climate conditions conducive to transmission. In this study, we developed a real-time PCR method for direct detection of two genomic regions of Haycocknema perplexum . The real-time PCR assays, SSU-PCR and COX1-PCR targeted the small subunit of nuclear ribosomal RNA and cytochrome oxidase-1 genomic regions, respectively. The performance of the assays was assessed using a panel of H. perplexum samples, both fresh frozen and formalin fixed paraffin embedded tissue (FFPE) (n = 22, derived from eight patients) and non- H. perplexum (n = 8) tissue biopsy specimens. Both H. perplexum assays showed 83% sensitivity and 100% specificity, with negative and positive predictive values of 100% and 93% respectively. The results indicate that the LOD was 10 −5 dilution (C t value 40.2) for COX1-PCR, and 10 −3 dilution (C t value 39.6) for SSU-PCR, making the latter a more sensitive assay for detecting lower concentrations of organism in the patient biopsy. The sensitivity of fresh frozen samples was superior to FFPE samples. All but 1 sample was negative following treatment. Feasibility of real-time PCR detection of H. perplexum directly from tissue biopsies has been demonstrated for diagnosis, possible test of cure and could enhance transmission surveillance.

BACKGROUND:Over the past 5 years, since publication of the initial review, studies have provided additional data on macrolide and fluoroquinolone resistance in Mycoplasma genitalium, including data from regions previously lacking this information. We aimed to provide contemporary estimates of macrolide and fluoroquinolone resistance in M genitalium to inform national, regional, and global treatment guidelines. METHODS:This is an update of a previous systematic review and meta-analysis, which was performed up to Jan 7, 2019. In this update, we searched PubMed, Embase, and MEDLINE from Jan 1, 2018, to April 18, 2023, for published studies reporting macrolide, fluoroquinolone, or dual-class (macrolide and fluoroquinolone) resistance in M genitalium. Data were combined with the previous meta-analysis to examine resistance prevalence in M genitalium samples collected up to and including 2021. Random-effects meta-analyses were used to calculate summary estimates of prevalence. Subgroup analyses by WHO region and four time periods (before 2012 to 2018-21) were performed. This study was registered with PROSPERO, number CRD42021273340. FINDINGS:166 studies (59 from the previous search period reporting data from M genitalium samples collected between 2003 and 2017, and 107 from the updated search period reporting data from M genitalium samples collected between 2005 and 2021) were included: 157 reporting macrolide resistance (41 countries; 22 974 samples), 89 reporting fluoroquinolone resistance (35 countries; 14 165 samples), and 74 reporting dual-class resistance (34 countries; 11 070 samples). In 2018-21, the overall prevalence of macrolide, fluoroquinolone, and dual-class resistance were 33·3% (95% CI 27·2-39·7), 13·3% (10·0-17·0), and 6·5% (4·0-9·4), respectively. Over time, there was a slight, although not statistically significant, decline in macrolide resistance in the Western Pacific and the Americas, but there was an increase in macrolide resistance in the European region. Fluoroquinolone resistance was highest in the Western Pacific and increased in the European non-Nordic region. ParC S83I was the most common variant associated with fluoroquinolone resistance, increasing from 0% (95% CI <0·0001-0·30) before 2012 to 7·3% (4·7-10·3) in 2018-21; ptrend=0·055. INTERPRETATION:Macrolide and fluoroquinolone resistance in M genitalium requires ongoing international surveillance, use of resistance assays for optimal antibiotic stewardship, and novel treatment options. FUNDING:Australian Research Council.

Background MPT64 is a key protein used for Mycobacterium tuberculosis (MTB) complex strain identification. We describe protracted transmission of an MPT64 negative MTB strain in Queensland, Australia, and explore genomic factors related to its successful spread. Methods All MPT64 negative strains identified between 2002 and 2022 by the Queensland Mycobacteria Reference Laboratory, and an additional 2 isolates from New South Wales (NSW), were whole genome sequenced. Bayesian modelling and phylogeographical analyses were used to assess their evolutionary history and transmission dynamics. Protein structural modelling to understand the putative functional effects of the mutated gene coding for MPT64 protein was performed. Findings Forty-three MPT64 negative isolates were sequenced, belonging to a single MTB cluster of Lineage 4.1.1.1 strains. Combined with a UK dataset of the same lineage, molecular dating estimated 1990 (95% HPD 1987–1993) as the likely time of strain introduction into Australia. Although the strain has spread over a wide geographic area and new cases linked to the cluster continue to arise, phylodynamic analysis suggest the outbreak peaked around 2003. All MPT64 negative strains had a frame shift mutation (delAT, p.Val216fs) within the MPT64 gene, which confers two major structural rearrangements at the C-terminus of the protein. Interpretation This study uncovered the origins of an MPT64 negative MTB outbreak in Australia, providing a richer understanding of its biology and transmission dynamics, as well as guidance for clinical diagnosis and public health action. The potential spread of MPT64 negative strains undermines the diagnostic utility of the MPT64 immunochromatographic test. Funding This study was funded from an operational budget provided to the Queensland Mycobacterium Reference Laboratory by Pathology Queensland, Queensland Department of Health.

Compared to cervical cancer, little is known about human papillomavirus (HPV)-driven oropharyngeal cancer and their cofactors. Here, we investigated potential associations between Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) with oral HPV and HPV persistence, which are known cofactors in cervical carcinogenesis, and also play a role in HPV-driven oropharyngeal cancer. Saliva samples (n = 547) from 312 people were tested for CT and NG and whom had previously been tested for oral HPV infection in a longitudinal study. Eight participants were positive for CT (2.6%) and one for NG (0.3%). Six of these nine participants were also positive for oral HPV in at least one of their samples. We found no significant associations between HPV, CT, or NG infection in the saliva samples analyzed. These preliminary data suggest CT and NG have little influence on oral HPV-positivity and persistence in a general population. However, larger studies focusing on 'at risk' population cohorts are necessary to assess potential associations between oral sexually transmissible infections and oral HPV infections, and their outcomes.

Antimicrobial resistance in Neisseria gonorrhoeae is an urgent global health threat. Ceftriaxone is the only remaining option for empirical monotherapy of gonococcal infections. The prevalence of ceftriaxone resistance in N gonorrhoeae is highest in the Western Pacific region.1Unemo M Lahra MM Escher M et al.WHO global antimicrobial resistance surveillance for Neisseria gonorrhoeae 2017–18: a retrospective observational study.Lancet Microbe. 2021; 2: e627-e636Summary Full Text Full Text PDF PubMed Google Scholar Many of the N gonorrhoeae infections associated with high minimum inhibitory concentrations (MICs) to ceftriaxone reported in the USA, the UK, and the rest of Europe carry the penA-60.001 allele, which is associated with ceftriaxone resistance and was first detected in the Western Pacific region.2Reimche JL Pham CD Joseph SJ et al.Novel strain of multidrug non-susceptible Neisseria gonorrhoeae in the USA.Lancet Infect Dis. 2024; 24: e149-e151Summary Full Text Full Text PDF Google Scholar, 3Day M Pitt R Mody N et al.Detection of 10 cases of ceftriaxone-resistant Neisseria gonorrhoeae in the United Kingdom, December 2021 to June 2022.Euro surveill. 2022; 272200803Crossref Scopus (25) Google Scholar, 6Maubaret C Caméléna F Mrimèche M et al.Two cases of extensively drug-resistant (XDR) Neisseria gonorrhoeae infection combining ceftriaxone-resistance and high-level azithromycin resistance, France, November 2022 and May 2023.Eurosurveillance. 2023; 282300456Crossref Scopus (4) Google Scholar The Enhanced Gonococcal Surveillance Programme in Cambodia recently reported 29 (38%) of 76 urethral isolates collected from 2021–22 were associated with ceftriaxone MICs of 0·125 mg/L or more.4Ouk V Pham CD Wi T van Hal SJ Lahra MM The Enhanced Gonococcal Surveillance Programme, Cambodia.Lancet Infect Dis. 2023; 23: e332-e333Summary Full Text Full Text PDF PubMed Google Scholar Moreover, all of those resistant isolates harbored the penA-60.001 allele. These findings are alarming and raise questions over whether ceftriaxone resistance and penA-60.001 are more widespread in the region. From January through to December, 2023, all isolates at the Hanoi Medical University Hospital (Hanoi, Viet Nam) were selected for antibiotic susceptibility testing (figure). MICs were interpreted through European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints. All isolates with ceftriaxone MICs of 0·125 mg/L or more underwent DNA extraction and subsequent penA-60.001 detection by real-time PCR, based on a penA-60.001 PCR previously used for enhanced surveillance in other settings.2Reimche JL Pham CD Joseph SJ et al.Novel strain of multidrug non-susceptible Neisseria gonorrhoeae in the USA.Lancet Infect Dis. 2024; 24: e149-e151Summary Full Text Full Text PDF Google Scholar, 5Whiley DM Mhango L Jennison AV Nimmo G Lahra MM Direct detection of penA gene associated with ceftriaxone-resistant Neisseria gonorrhoeae FC428 strain by using PCR.Emerg Infect Dis. 2018; 24: 1573-1575Crossref PubMed Scopus (20) Google Scholar In total, 243 N gonorrhoeae isolates were cultured and 13 (5%) of these were non-viable. Of the 230 available isolates, 224 (97%) were obtained from urethral specimens. Ceftriaxone MICs greater than 0·125 mg/L were found in 64 (28%) of the 230 isolates and MICs greater than 0·25 mg/L were detected in 50 (22%) isolates. The penA-60.001 positivity was 67% (n=39/64) among those with MICs greater than 0·125 mg/L and 62% (n=31/50) among isolates with MICs greater than 0·25 mg/L. All (n=64/64) isolates with ceftriaxone MICs greater than 0·125 mg/L were resistant to cefixime, with MICs of 0·5 mg/L or more, and were susceptible to azithromycin (EUCAST epidemiological cutoff ≤1 mg/L). These findings highlight the troublingly high prevalence of ceftriaxone resistance in the Western Pacific region, with more than a quarter of N gonorrhoeae isolates resistant to first-line therapy in this setting, and that strains harboring penA-60.001 are becoming increasingly prevalent in many settings in the Western Pacific region. Our report underscores the utility of direct detection of penA-60.001, which could be used to rapidly detect resistance and might prove to be a useful surveillance tool more widely. The findings also suggest the presence of non-penA-60.001 mutations contributing to ceftriaxone resistance in this setting. Further analyses with whole-genome sequencing and multilocus sequence typing would help to elucidate those resistance mutations and better understand their transmission dynamics. Ceftriaxone as first-line therapy for gonorrhea is under threat and interventions are urgently needed to improve the detection of resistance and to mitigate further transmission. DMW reports research funding from SpeeDx, outside the submitted work. All other authors declare no competing interests. This work was supported by the National Institutes of Health, Fogarty International Center (K01TW012170 to PCA). The funder had no role in the data collection, analysis, or interpretation, or the decision to submit to publication.