The LightMix (R) Modular Mycoplasma Macrolide and LightMix (R) Modular parC Fluoroquinolone Resistance assays (TIB Molbiol) were evaluated using sequential Mycoplasma genitalium positive (n = 125) and negative (n = 93) clinical samples. Results were compared to the results of an established commercial assay (ResistancePlus MG assay, SpeeDx Pty Ltd) or Sanger sequencing (for parC). Detection of M. genitalium by the TIB Molbiol assay had a high agreement with the reference assay, with a positive percent agreement (PPA) of 97.6 [95% confidence interval (CI): 93.1-99.5] and negative percent agreement (NPA) of 95.7 (95% CI: 89.5-98.8). From 105 positive samples, macrolide resistance detection had a PPA of 100% (95% CI: 93.7-100) and NPA of 81.3% (95% CI: 67.4-91.1). For the detection of fluroquinolone resistance mutation G248T/S83I or "other mutation" in the quinolone resistance determinant region, from 95 samples there was 100% (95% CI: 86.3-100) sensitivity and 100% (95% CI: 94.5-100) specificity. The understanding of the basis for fluoroquinolone treatment failure is still developing; it is therefore important to use the output of parC-based resistance assays with caution to avoid the inappropriate use of antibiotic therapies, especially considering the limited number of alternative treatments.

Background: In Australia, high and widespread uptake of the quadrivalent human papillomavirus (HPV) vaccine has led to substantial population-level reductions in the prevalence of quadrivalent vaccine targeted HPV genotypes 6/11/16/18 in women aged <= 35 years. We assessed risk factors for HPV detection among 18-35 year old women, 9-12 years after vaccine program introduction. Methods: Women attending health services between 2015 and 2018 provided a self-collected vaginal specimen for HPV genotyping (Roche Linear Array) and completed a questionnaire. HPV vaccination status was validated against the National Register. Adjusted odds ratios (aORs) and 95% confidence intervals (CI) were calculated for factors associated with HPV detection. Results: Among 1564 women (median age 24 years; IQR 21-27 years), Register-confirmed >= 1-dose vaccine coverage was highest at 69.3% and 68.1% among women aged 18-21 and 22-24 years respectively, decreasing to 42.9% among those aged 30-35 years. Overall prevalence of quadrivalent vaccine-targeted HPV types was very low (2.0%; 95% CI: 1.4-2.8%) and influenced only by vaccination status (5.5% among unvaccinated compared with 0.7% among vaccinated women; aOR = 0.13 (95% CI: 0.05-0.30)). Prevalence of remaining HPV types, at 40.4% (95% CI: 38.0-42.9%), was influenced by established risk factors for HPV infection; younger age-group (p-trend < 0.001), more recent (p < 0.001) and lifetime sexual partners (ptrend < 0.001), but not vaccination status. Prevalence of HPV31/33/45, which shared risk factors with that of non-vaccine targeted HPV types, was also lower among vaccinated (4%) compared with unvaccinated (7%) women (aOR = 0.51; 95% CI: 0.29-0.89), indicative of cross-protection. Conclusion: Vaccination has changed the epidemiology of HPV infection in Australian women, having markedly reduced the prevalence of vaccine-targeted types, including amongst women with known risk factors for infection. Vaccinated women appear to be benefiting from modest cross-protection against types 31/33/45 afforded by the quadrivalent HPV vaccine. These results reinforce the importance of HPV vaccination. (C) 2021 Published by Elsevier Ltd.

Background Men-who-have-sex-with-men (MSM) living with HIV have a high prevalence of high-risk human papillomavirus (hrHPV) in the anus, and a high incidence of anal cancer. We conducted an open-label, single-arm pilot study to assess the utility of imiquimod cream against hrHPV among MSM living with HIV.[ACTRN12617001355369] Methods The study was conducted at Melbourne Sexual Health Centre between April 2018 and June 2020. MSM aged ≥18 years, living with HIV, who tested positive for any anal hrHPV on clinician-collected swabs were eligible. We instructed men to apply 5% imiquimod cream (6.25 mg) intra-anally and peri-anally 3 doses per-week for 16-weeks (Phase 1), followed by a maintenance period of 1 dose per-week for 48-weeks (Phase 2). We collected adverse events (AE) using text messages and questionnaires. Results Thirty MSM were enrolled to phase 1 and 27 completed the week 16 follow-up(median age 50). Twenty-four MSM (86%) applied at least 50% of imiquimod doses. All men reported AE, with 39.5% reporting grade 1,39.5% grade 2, and 21% grade 3 AEs. Eighteen MSM (67%) required treatment interruption. The reasons for interruption were haemorrhoids (n=3), herpes genitalis reactivation (n=2), and grade 2–3 AEs (n=11) such as irritation, itching and tenderness. 60% of MSM with grade 2(n=6) and 33% with grade 3 AEs (n=2) opted to continue to phase 2. Eighteen MSM were enrolled to phase 2 and 13 completed the week 48 follow-up. All MSM took at least 50% of doses. No treatment-limiting AEs were reported. 10 MSM in phase 1 (37%) and none in phase 2 reported their sex life was negatively impacted from imiquimod use. Conclusion Intra-anal and peri-anal imiquimod at 3 doses per-week was poorly tolerated over 16 weeks and most men required treatment interruption due to AEs. In contrast, once-a-week application was well tolerated with no treatment-limiting AEs reported over 48-weeks.

Men who have sex with men (MSM) living with HIV have a high prevalence and incidence of anal high-risk human papillomavirus (hrHPV) and anal cancer. We conducted an open-label, single-arm pilot study to examine the tolerability of imiquimod cream among MSM aged ≥18 years, living with HIV, who tested positive for anal hrHPV at Melbourne Sexual Health Centre between April 2018 and June 2020. We instructed men to apply 6.25 mg imiquimod intra-anally and peri-anally 3 doses per week for 16 weeks (period 1) and then one dose per week for a further 48 weeks (period 2). Twenty-seven MSM enrolled in period 1 and 24 (86%) applied at least 50% of doses. All men reported adverse events (AEs), including 39.5% grade 1, 39.5% grade 2, and 21% grade 3 AEs on at least one occasion. Eighteen MSM (67%) temporarily stopped using imiquimod during period 1, most commonly due to local AEs (n = 11) such as irritation and itching. Eighteen MSM continued in period 2 and all applied at least 50% of doses with no treatment-limiting AEs reported. Imiquimod 3 doses per week caused local AEs in most men and was not well tolerated. In contrast, once-a-week application was well tolerated over 48-weeks with no treatment-limiting AEs.

Background Anal infection with high-risk human papillomavirus (HPV) genotypes 16 and 18 and anal cancer are overrepresented in men who have sex with men (MSM). This study investigated HPV prevalence in young MSM before and after the implementation of a school-based quadrivalent HPV (genotypes 6, 11, 16, and 18) vaccination programme for boys in Australia in 2013. Methods In this repeated cross-sectional study, MSM aged 16-20 years were recruited from two successive birth cohorts via sexual health clinics and the community in Melbourne, Australia. The first cohort was before the implementation of gender-neutral vaccination (HYPER1 study, done in 2010-12, NCT01422356), and the second was the post-vaccination cohort (HYPER2 study, done in 2017-18, NCT03000933). Men who self-identified as being same-sex attracted were enrolled, and those recruited via the HYPER2 study had to be resident in Australia since 2013 to ensure eligibility. Study procedures were done in the Melbourne Sexual Health Centre. A clinician-collected anal swab and self-collected penile swab and oral rinse were tested for 28 HPV genotypes, and data on demographics and sexual health practices were collected via questionnaires. Only assessable samples were included in the analyses. We compared anatomical site-specific prevalence of HPV genotypes between cohorts by calculating the prevalence ratio, adjusting for age, circumcision, and sex with women. Herd protection was also assessed, by calculating the adjusted prevalence ratios by vaccination status. Findings 400 MSM, 200 per cohort, were included in the study. In both cohorts, the median number of lifetime male partners was ten (IQR 5-25). The prevalence of any anal quadrivalent vaccine-preventable HPV genotype was higher in the pre-vaccination cohort (54 [28%] of 193) than in the post-vaccination cohort (14 [7%] of 193; adjusted prevalence ratio [PR] 0middot24, 95% CI 0middot14-0middot42), largely driven by decreases in HPV6, followed by HPV11, 16, and 18. Nevertheless, there was also a significant reduction in anal HPV16 and 18 in the post-vaccination cohort from the pre-vaccination cohort (0middot31, 0middot14-0middot68). The prevalence of any penile quadrivalent vaccine-preventable HPV genotype was also higher in the pre-vaccination cohort (21 [12%] of 177) than in the post-vaccination cohort (11 [6%] of 179; 0middot48, 0middot24-0middot97), driven by decreases in HPV 6 and 11, but not by 16 and 18. The prevalence of any oral quadrivalent vaccine-preventable HPV genotype was higher in the pre-vaccination cohort (seven [4%] of 200) than in the post vaccination cohort (one [1%] of 199; 0middot10, 0middot01-0middot97); there were no cases of oral HPV6 or 11 detected in HYPER2. Comparing the pre-vaccinated cohort with the 149 confirmed vaccinated men from HYPER2 showed a reduction in any quadrivalent vaccine-preventable HPV genotype for anal (0middot09, 0middot03-0middot25) and penile (0middot18, 0middot05-0middot59) infection but not for oral infection (0middot17, 0middot03-1middot08). Interpretation A reduction in anal, penile, and oral quadrivalent vaccine-targeted genotypes occurred in young MSM following the implementation of a school-based gender-neutral HPV vaccination programme. The fall in anal HPV16 and 18 may lead to a reduction in the incidence of anal cancer. Funding Merck and the Australian Government Department of Health. Copyright (c) 2021 Elsevier Ltd. All rights reserved.

Antibody levels are substantially higher following vaccination against human papillomavirus types 6, 11, 16, and 18 compared with natural infection among adolescent gay and bisexual men aged 16-20 years. Background Australia introduced a school-based gender-neutral human papillomavirus (HPV) vaccination program for girls and boys aged 12-13 years in 2013. We examined HPV type-specific antibody levels in unvaccinated young men who have sex with men (MSM) with natural infection and compared these with levels in those vaccinated against HPV. Methods Serum specimens at baseline were collected from MSM aged 16-20 years in the HYPER1 (Human Papillomavirus in Young People Epidemiological Research) and HYPER2 studies, conducted in 2010-2013 and 2017-2019, respectively. Merck's 4-plex HPV competitive Luminex Immunoassay was used to quantify HPV6-, HPV11-, HPV16-, and HPV18-specific antibodies. We compared antibody levels for each HPV genotype between unvaccinated men (HYPER1) and vaccinated men (HYPER2) using the Mann-Whitney U test. Results There were 200 unvaccinated men and 127 vaccinated men included in the analysis. Median antibody levels among vaccinated men were significantly higher than levels among unvaccinated men for HPV6 (223 milli-Merck units per milliliter [mMU/mL] vs 48 mMU/mL, P < .0001), HPV11 (163 mMU/mL vs 21 mMU/mL, P < .0001), HPV16 (888 mMU/mL vs 72 mMU/mL, P < .0001), and HPV18 (161 mMU/mL vs 20 mMU/mL, P < .0001). Antibody levels did not change over time for up to 66 months for all 4 genotypes among vaccinated men. Conclusions Among young MSM vaccinated with the quadrivalent HPV vaccine, antibody levels for HPV6, HPV11, HPV16, and HPV18 were significantly higher than those in unvaccinated MSM following natural infection. Antibody levels following vaccination appeared to remain stable over time.

Aim Validate the Roche, MagNAPure96 (MP96) nucleic acid extraction platform for Seegene Anyplex II HPV28 (Anyplex28) detection of Human Papillomavirus. Methods and Results Comparisons were made for Anyplex28 genotyping from 115 cervical samples extracted on the Hamilton, STARlet and the MP96. Two DNA concentrations were used for the MP96, one matched for sample input to the STARlet and another 5x concentration (laboratory standard). Agreement of HPV detection was 89 center dot 8% (kappa = 0 center dot 798; P = 0 center dot 007), with HPV detected in 10 more samples for the MP96. There was a high concordance of detection for any oncogenic HPV genotype (kappa = 0 center dot 77; P = 0 center dot 007) and for any low-risk HPV genotype (kappa = 0 center dot 85; P = 0 center dot 008). DNA extracted at laboratory standard had a lower overall agreement 85 center dot 2% (kappa = 0 center dot 708; P < 0 center dot 001), with 17/115 discordant positive samples that tested negative after STARlet extraction. Of the discordant genotypes, 72 center dot 7% were detected in the lowest signal range for Anyplex28 ('+'). Conclusions MP96 performed with high concordance to STARlet, although produced DNA with a higher analytical sensitivity on the Anyplex28. Significance and Impact of the Study This analysis supports the use of samples extracted on the MP96 for HPV genotyping using the Anyplex28. Furthermore, an increase in DNA concentration increased analytical sensitivity of the Anyplex28, particularly appropriate for prevalence studies.

Background Australia introduced a school-based quadrivalent human papillomavirus (HPV) vaccination program for females in 2007. This was extended to include boys aged 12–13 from 2013, with a two-year catch-up for boys aged ≤15. This study examined HPV prevalence among young gay and bisexual males (GBM) who were age-eligible for vaccination in the school-based program. Methods Males aged 16–20 years were recruited from sexual health clinics and the community in Melbourne in 2017–2018, if they reported any form of male sexual contact, and were residents of Australia from 2013. A clinician-collected anal swab, self-collected penile swab and oral rinse were collected and analysed for detection and 37 HPV genotypes (Roche Linear Array). Preliminary results from 114 GBM were analysed and full results will be available for presentation. Results The mean age of GBM was 18.6 years (SD 1.0). The majority (80%) were recruited from clinics and 20% from the community. The median number of lifetime male partners was 10 [IQR 5–25] for receptive oral sex, four [IQR 1–11] for receptive anal sex and one for insertive anal sex [IQR 0–6]. Overall, 64% received at least one dose of vaccine documented via the National HPV Vaccination Program Register. Prevalence of quadrivalent vaccine-preventable HPV genotypes was 4.9% (95% CI: 1.6–11%) for anal, 3.4% (95% CI: 0.7–9.5%) for penile and 0% (95% CI: 0–3.2%) for oral sites. Only two men, both unvaccinated, had high-risk vaccine-preventable HPV genotypes: one with anal HPV16 (1%); the other penile HPV16 (1%). Conclusion Statistical analysis comparing before and after the male vaccination program will be performed until recruitment is completed. The preliminary analysis shows the prevalence of anal HPV 16/18 among young GBM following the school-based male HPV vaccination was low. The addition of male HPV vaccination to female programs may reduce the incidence of anal cancer among GBM. Disclosure No significant relationships.

Background Australia introduced a school-based human papillomavirus (HPV) vaccination program for females aged 12–13 years in 2007, with a three-year catch-up to age 26; and for boys aged 12–13 from 2013, with a two-year catch-up to age 15. This study aimed to compare the prevalence of penile HPV between teenage heterosexual males in cohorts eligible or non-eligible for the school-based male vaccination program. Methods Between 2014 and 2017, sexually active heterosexual males aged 17–19 were recruited from sexual health centres and community sources across Australia. Males provided a self-collected penile swab for HPV genotypes (Roche Linear Array) and completed a questionnaire. HPV prevalence was compared between males in two periods: 2014–2015 (non-eligible for school-based male vaccination) and 2016–2017 (eligible for school-based male vaccination). Self-reported vaccine doses were confirmed with doses reported to the National HPV Vaccination Program Register. Results Overall, 152 males were recruited in 2014–2015 and 146 in 2016–2017. Numbers of female sex partners and condom use did not differ between the two periods. Prevalence of quadrivalent vaccine-preventable [4vHPV] genotype (6, 11, 16, 18) was low in both periods (2.6% in 2014–2015 versus 0.7% in 2016–2017; p=0.37). Compared with males recruited in 2014–2015, males in 2016–2017 had a lower prevalence of: any 37 HPV genotype (21.7% versus 11.6%; p=0.02); and any 13 high-risk genotype (15.8% versus 7.5%; p=0.03). Prevalence of low-risk HPV genotypes did not differ between the two periods (p=0.25). Of the males recruited in 2016–2017, 55% had received ≥1 vaccine dose. Conclusion Prevalence of 4vHPV genotypes among teenage heterosexual males in both cohorts was low, presumably due to herd protection from the female-only vaccination program. The addition of the school-based male vaccination was associated with a lower prevalence of high-risk HPV genotypes other than genotypes 16/18. Disclosure No significant relationships.

Background In Australia, high and widespread uptake of the quadrivalent human papillomavirus (HPV) vaccine has led to substantial population-level reductions in the prevalence of HPV16/18 in women aged ≤35 years. We assessed risk factors for oncogenic HPV detection among 18–35 year old women in 2015–2018. Methods Women attending health services across Australia provided a self-collected (vaginal) or clinician-collected (cervical) specimen for HPV genotyping (Roche Linear Array) and completed a questionnaire. HPV vaccination status was validated against the National HPV Vaccination Program Register. Odds ratios (ORs) and 95% confidence intervals (CI) were calculated for factors associated with detection of any oncogenic HPV (HPV16/18/31/33/35/39/45/51/52/56/58/59/66/68). Results Among 1,643 women, vaccine coverage (≥one dose) was 61.5% (69.1%, 58.7% and 41.1% among those 18–24, 25–29 and 30+ years, respectively). Oncogenic HPV prevalence was 25.4% (95% CI: 23.2–27.6%). In univariable analysis, risk factors for detection included younger age (p-trend<0.001), being a current smoker (p=0.05), and reporting more lifetime (p-trend<0.001) and recent (last 12 months) sexual partners (p-trend<0.001). In multivariable analysis, younger age (adjusted OR=1.33 [1.17–1.52]), more lifetime (adjusted OR=1.33 [1.16–1.52]) and recent sexual partners (adjusted OR=2.37 [1.77–3.16]) remained significant. There were no associations with socioeconomic status, area of residence or vaccination history. HPV16/18 prevalence was 1.4% (0.9–2.1%). In univariable analysis, risk factors for detection included older age (p=0.05), being non-Australian born (p=0.05), and being unvaccinated (p<0.001). In multivariable analysis, being unvaccinated remained the only factor significantly associated with HPV16/18 detection (adjusted OR=8.61 [2.45–30.25]). There were no associations with area of residence, socioeconomic status, or sexual behaviour. Conclusion Oncogenic HPV was commonly detected among young Australian women; prevalence was influenced by risk factors related to sexual behaviour. In contrast, prevalence of quadrivalent vaccine-targeted types 16/18 was very low and influenced only by vaccination status. Vaccination has changed the epidemiology of HPV infection in Australia. Disclosure No significant relationships.

Background: Australia introduced a school-based human papillomavirus (HPV) vaccination program for females aged 12-13 years in 2007, with a three-year catch-up to age 26; and for boys aged 12-13 from 2013, with a two-year catch-up to age 15. This study aimed to compare the prevalence of penile HPV between teenage heterosexual males in cohorts eligible or non-eligible for the school-based male vaccination program. Methods: Between 2014 and 2017, sexually active heterosexual males aged 17-19 were recruited from sexual health centres and community sources across Australia. Males provided a self-collected penile swab for 37 HPV genotypes using Roche Linear Array and completed a questionnaire. We calculated adjusted prevalence ratios (aPR) of HPV between males in two periods: 2014-2015 (preceding implementation of school-based male vaccination) and 2016-2017 (eligible for school-based male vaccination). Self-reported vaccine doses were confirmed with doses reported to the National HPV Vaccination Program Register. Results: Overall, 152 males were recruited in 2014-2015 and 146 in 2016-2017. Numbers of female sex partners and condom use did not differ between the two periods. The prevalence of quadrivalent vaccine-preventable [4vHPV] genotypes (6/11/16/18) was low in both periods (2.6% [2014-15] versus 0.7% [2016-17]; p = 0.371; aPR 0.28 [95% CI: 0.03-2.62]). Compared with men in 2014-2015, men in 2016-2017 had a lower prevalence of any of the 37 HPV genotypes tested (21.7% versus 11.6%; aPR 0.62 [95% CI: 0.36-1.07]) and any of the 13 high-risk genotypes tested (15.8% versus 7.5%; aPR 0.59 [95% CI: 0.30-1.19]). Prevalence of low-risk HPV genotypes did not differ between the two periods. Of the males recruited in 2016-2017, 55% had received >= 1 vaccine dose. Conclusion: The prevalence of 4vHPV genotypes among teenage heterosexual males in both cohorts was low, presumably due to herd protection from the female-only vaccination program. Further studies are required to determine the impact of universal HPV vaccination on HPV prevalence in males. (C) 2019 Elsevier Ltd. All rights reserved.

Introduction: Australia has recently implemented major changes in cervical cancer prevention policies including introduction of primary human papillomavirus (HPV) screening starting at age 25, and replacement of the quadrivalent HPV vaccine with the nonavalent vaccine in the national school-based program. We assessed the feasibility and utility of conducting HPV testing in residual clinical specimens submitted for routine Chlamydia trachomatis screening, as a means of tracking HPV vaccine program impact among young sexually active women. Methods: De-identified residual specimens from women aged 16-24 years submitted for chlamydia testing were collected from three pathology laboratories in Victoria and New South Wales. Limited demographic information, and chlamydia test results were also collected. Patient identifiers were sent directly from the laboratories to the National HPV Vaccination Program Register, to obtain HPV vaccination histories. Samples underwent HPV genotyping using Seegene Anyplex II HPV 28 assay. Results: Between April and July 2018, 362 residual samples were collected, the majority (60.2%) of which were cervical swabs. Demographic data and vaccination histories were received for 357 (98.6%) women (mean age 21.8, SD 2.0). Overall, 65.6% of women were fully vaccinated, 9.8% partially, and 24.7% unvaccinated. The majority (86.0%) resided in a major city, 35.9% were classified in the upper quintile of socioeconomic advantage and chlamydia positivity was 7.8%. The prevalence of quadrivalent vaccine-targeted types (HPV6/11/16/18) was 2.8% (1.5-5.1%) overall with no differences by vaccination status (p = 0.729). The prevalence of additional nonavalent vaccine-targeted types (HPV31/33/45/52/58) was 19.3% (15.6-23.8%). One or more oncogenic HPV types were detected in 46.8% (95% CI 41.6-52.0%) of women. Conclusions: HPV testing of residual chlamydia specimens provides a simple, feasible method for monitoring circulating genotypes. Applied on a larger scale this method can be utilised to obtain a timely assessment of nonavalent vaccine impact among young women not yet eligible for cervical screening. (C) 2019 Elsevier Ltd. All rights reserved.