Use of genome sequencing to resolve differences in gradient diffusion and agar dilution antimicrobial susceptibility testing performance of Neisseria gonorrhoeae isolates in Alberta, Canada.

Agar dilution is the gold standard method for phenotypic antimicrobial susceptibility testing (AST) for Neisseria gonorrhoeae. However, this method is laborious and requires expertise, so laboratories that perform N. gonorrhoeae AST may choose alternative methods such as disk diffusion and gradient diffusion. In this study, we retrospectively compare the performance of gradient diffusion to agar dilution for 2,394 unique N. gonorrhoeae isolates identified in Alberta from 2017 to 2020 against azithromycin, cefixime, ceftriaxone, ciprofloxacin, penicillin, and tetracycline. Genome sequencing was utilized to resolve discrepancies between AST methods, detect antimicrobial resistance markers, and identify trends between error rates and sequence types (STs) of isolates. Over 90% of N. gonorrhoeae isolates were susceptible to azithromycin, cefixime, and ceftriaxone, whereas decreased susceptibility was observed for ciprofloxacin, penicillin, and tetracycline. Categorical (CA) and essential agreement (EA) was poorest between the two methods for penicillin (CA: 86.02%; EA: 77.69%) and tetracycline (CA: 47.22%; EA: 55.96%); however, the low CA was primarily attributed to minor errors. Antimicrobial agents with errors outside of acceptable limits included azithromycin (very major error: 18.42%; major error: 7.73%) and tetracycline (very major error: 6.17%). Genome sequencing on a subset of isolates resolved 30.3% of the azithromycin major errors and confirmed the azithromycin or tetracycline very major errors. Significant associations between certain STs and error types for azithromycin and tetracycline were also identified. Overall, gradient diffusion compared well to agar dilution for cefixime, ceftriaxone, and ciprofloxacin, and genome sequencing was identified as a useful tool to arbitrate discrepant susceptibility testing results between gradient diffusion and agar dilution for N. gonorrhoeae.

Case Series of Stillbirths Due to Syphilis in Edmonton, Alberta, Canada.

BACKGROUND: Data on the incidence and characteristics of stillbirths attributed to congenital syphilis were collected. METHODS: We extracted data on stillbirths in the Edmonton Zone on January 1, 2015, through June 30, 2021, born to persons diagnosed with infectious syphilis (primary, secondary, early latent, or early neurosyphilis) during pregnancy or at the time of delivery. RESULTS: Of 314 infants documented to be exposed to infectious syphilis during gestation, 16 (5.1%) were stillborn. Three of the 16 females with stillbirths were diagnosed with syphilis during pregnancy but not treated, 12 were diagnosed only at the time of stillbirth (1 of whom was treated early in pregnancy and presumably reinfected), and 1 had a stillbirth in the week after one dose of benzathine penicillin G. CONCLUSIONS: Stillbirths due to congenital syphilis were all due to failure to treat syphilis in pregnancy. Innovative strategies to prevent syphilis in the community and to reach those experiencing barriers to care are urgently required to not miss opportunities to diagnose and treat syphilis as early as possible during pregnancy.

Serologic follow-up of solid organ transplant recipients who received organs from donors with reactive syphilis tests: A retrospective cohort study.

The increased procurement of organs from donors with risk factors for blood-borne diseases and the expanding syphilis epidemic have resulted in a growing number of organs transplanted from donors with reactive syphilis serology in our center. Based on guidelines, recipients typically receive therapy shortly after the transplant, but data on outcomes are limited. The primary objective of this study was to determine syphilis seroconversion rates at three months post-transplant in recipients of solid organs procured from donors with reactive syphilis serology. Organ donors and recipients were tested for syphilis antibody; positive results were confirmed with Treponema pallidum Particle Agglutination (TPPA). Eleven donors with reactive syphilis antibody donated organs to 25 syphilis negative recipients. Three recipients seroconverted at post-transplant month 3. All of them had received therapy shortly after transplant. TPPA was negative in all 3. Despite post-transplant treatment, 3 of 25 (12%) syphilis negative recipients of organs from syphilis positive donors seroconverted at 3 months. All remained TPPA negative possibly reflecting passive antibody transfer or differing test sensitivity to low level treponemal antibodies. Further studies are needed to assess optimal syphilis transmission prevention strategies and follow up recipient testing in organ transplantation.

Retrospective Cohort Study Examining the Correlates of Reported Lifetime Stimulant Use in Persons Diagnosed With Infectious Syphilis in Alberta, Canada, 2018 to 2019.

BACKGROUND: We sought to examine the correlates for stimulant use in persons diagnosed with infectious syphilis during an outbreak in Alberta to help guide public health interventions. METHODS: Infectious syphilis data were extracted from the Communicable Disease and Outbreak Management database from January 1, 2018, to December 31, 2019. Behavioral, demographic, and lifetime reported stimulant use data were obtained. Descriptive analyses and logistic regression were performed for 3 subpopulations (gay, bisexual, and other men who have sex with men; men who have sex with women; and women). RESULTS: Of 3627 individuals diagnosed with infectious syphilis, 23.9% (n = 867) cases were not interviewed for substance use and were removed from further analysis. Of the remaining 2759 people, 41.8% (n = 1153) self-reported lifetime stimulant use. Gay, bisexual, and other men who have sex with men reported stimulant use less often than women (24.6% vs. 44.1%; P < 0.0001) and men who have sex with women (24.6% vs. 46.2%; P < 0.0001). Multivariable analyses demonstrated that stimulant use was associated with persons who injected drugs, had correctional involvement, or reported multiple sex partners. Men who have sex with women were more likely to self-report First Nations ethnicity (adjusted odds ratio, 1.76 [95% confidence interval, 1.25-2.49]), and women were more likely to have a concurrent gonorrhea infection (adjusted odds ratio, 1.62 [95% confidence interval, 1.15-2.28]). CONCLUSIONS: Nearly half of infectious syphilis cases in Alberta reported lifetime nonprescription stimulant use. Infectious syphilis cases with stimulant use were associated with injection drug use, multiple sex partners, and correctional involvement. Our observations highlight the need for integration of sexual health services into programs for people who use substances and those in corrections custody.

Retrospective Cohort Study of the Incidence and Outcomes of Jarisch-Herxheimer Reactions After Treatment of Infectious Syphilis in Late Pregnancy.

ABSTRACT: Of 39 pregnant women at ≥20 weeks' gestation treated with benzathine penicillin G for infectious syphilis, we identified only 2 mild Jarisch-Herxheimer reactions. There were no immediate fetal sequelae. Data from our study do not support the recommendation for routine admission for the treatment of infectious syphilis in late pregnancy.

Molecular Surveillance and Prediction of Antimicrobial Resistance of Neisseria gonorrhoeae in Northern Alberta, Canada, 2015 to 2018.

BACKGROUND: The aims of this study was to describe molecular surveillance of Neisseria gonorrhoeae in the North Zone of Alberta (NZ) and to determine its value in predicting antimicrobial resistance. METHODS: Sequence types (STs) and single-nucleotide polymorphism (SNP) assays were performed on nucleic acid amplification testing (NAAT) samples. Sequence types of NAATs were matched to ST of cultures from across Alberta. Antimicrobial resistance prediction of NAATs for cephalosporins, azithromycin, and ciprofloxacin using SNP was compared with matching ST culture results using agar dilution and whole-genome sequencing. RESULTS: Of 2755 eligible specimens (2492 cases), 61.9% (1646 specimens) were sent for sequence typing, identifying 196 unique ST. Antimicrobial resistance data for 1307 additional cases were available using matching cultures. Decreased susceptibility (DS) to antimicrobials used for gonorrhea treatment was rare in the NZ; according to the SNP assay, none of the specimens had predicted DS to cephalosporins or azithromycin resistance. However, of the NZ NAAT samples tested in this study, 10.7% (131 of 1220) were predicted to have intermediate cephalosporin minimum inhibitory concentrations and 9.6% (115 of 1204) were resistant to ciprofloxacin. Based on cultures, the proportions of resistance in all of Alberta were as follows: DS to cephalosporins, 0.6% (20 of 3373); DS to intermediate cephalosporin, 16.9% (570 of 3373); azithromycin resistance, 1.2% (41 of 3373); and ciprofloxacin resistance, 32.2% (1087 of 3373). CONCLUSIONS: Our results highlight our ability to use culture-independent methods to predict antimicrobial resistance in N. gonorrhoeae.

Examination of Care Milestones for Preventing Congenital Syphilis Transmission Among Syphilis-Infected Pregnant Women in Alberta, Canada: 2017-2019.

BACKGROUND: An infectious syphilis outbreak in Alberta has resulted in increased congenital syphilis (CS) cases. To shed light on potential risk factors, we used administrative data sets to examine care milestones for the prevention of CS among pregnant women diagnosed with syphilis, as well as correlates of women giving birth to infants with CS. METHODS: Provincial administrative databases were used to identify and describe pregnant women diagnosed with any stage of infectious or noninfectious syphilis who gave birth in Alberta between January 1, 2017, and December 31, 2019. Data on prenatal care, syphilis screening, and syphilis medication dispensation were used to evaluate the care milestones. Clinical care and maternal demographics were assessed using logistic and linear regression analyses to determine correlates for missed care milestones or a newborn outcome of CS. RESULTS: Of 182 syphilis-infected pregnant women, 63 (34.6%) delivered a newborn with CS. Overall, in the first trimester, 136 (75.1%) women had a health care visit, 72 (39.6%) had a prenatal care visit, 71 (39.0%) were screened for syphilis, and 44 (24.2%) were treated. Gestational time to treatment initiation (adjusted odds ratio, 1.04; 95% confidence interval, 1.02-1.06) and older maternal age at diagnosis (adjusted odds ratio: 1.28, 95% confidence interval, 1.08-1.50) were independently associated with CS outcomes. No variables were found to be independently associated with a health care visit, prenatal screening, or initiation of treatment. CONCLUSIONS: Although nearly two-thirds of CS cases were prevented, there remained missed opportunities in the prevention of CS. Early treatment, which relies on timely access to prenatal care and screening, was the most important for the prevention of CS.

Expanding access to HIV testing through Canadian community pharmacies: findings from the APPROACH study.

BACKGROUND: There is a need for acceptable and feasible HIV testing options to ensure people living with HIV know their status so they can access care. Pharmacist-provided HIV point-of-care testing (POCT) may overcome testing barriers, including privacy concerns, testing wait times, and improve accessibility. In the APPROACH study, we aimed to develop and assess an HIV POCT program in community pharmacies for future scale up and evaluation. This paper describes the program uptake, participant and pharmacist experiences, and implementation factors. METHODS: A pharmacist-provided HIV POCT program was offered in 4 pharmacies in two Canadian provinces. A mixed methods design incorporated self-report questionnaire data, participant telephone interviews, pharmacist focus groups, workload analysis, and situational analysis to assess the uptake, acceptability and feasibility of the HIV POCT program. RESULTS: Over the 6-month pilot, 123 HIV tests were performed. One new case of HIV was identified; this participant was linked with confirmatory testing and HIV care. Participants were predominantly male (76%), with a mean age of 35 years. This was the first HIV test for 27% participants, and 75% were at moderate to very high risk of undiagnosed HIV infection, by Denver HIV Risk Score. Questionnaires and telephone interviews showed participants were very satisfied with the program; 99% agreed HIV POCT should be routinely offered in pharmacies and 78% were willing to pay for the service. Participants felt the pharmacy was convenient, discreet, and that the pharmacist was supportive and provided education about how to reduce their future risk. Pharmacists felt prepared, confident, and expressed professional satisfaction with offering HIV POCT. Community and public health supports, clear linkage to care plans to refer participants with positive HIV POCT results, and provision of counselling tools were important enabling factors for the program. Pharmacist remuneration, integration with existing healthcare systems, and support for ongoing promotion of HIV POCT availability in pharmacies were identified as needs for future scale-up and sustainability. CONCLUSIONS: A successful model of pharmacy-based POCT, including linkage to care, was developed. Further research is needed to determine the effectiveness and cost-effectiveness of this approach in finding new diagnoses and linking them with care. TRIAL REGISTRATION: Retrospectively registered with clinicaltrials.gov (NCT03210701) on July 6, 2017.

Genetic Characterization and Enhanced Surveillance of Ceftriaxone-Resistant Neisseria gonorrhoeae Strain, Alberta, Canada, 2018.

In July 2018, a case of Neisseria gonorrhoeae associated with ceftriaxone treatment failure was identified in Alberta, Canada. We identified the isolate and nucleic acid amplification testing (NAAT) specimen as the ceftriaxone-resistant strain multilocus sequence type 1903/NG-MAST 3435/NG-STAR 233, originally identified in Japan (FC428), with the same penA 60.001 mosaic allele and genetic resistance determinants. Core single-nucleotide variant (SNV) analysis identified 13 SNVs between this isolate and FC428. Culture-independent surveillance by PCR for the A311V mutation in the penA allele and N. gonorrhoeae multiantigen sequence typing directly from NAAT transport media positive for N. gonorrhoeae by NAAT did not detect spread of the strain. We identified multiple sequence types not previously detected in Alberta by routine surveillance. This case demonstrates the benefit of using culture-independent methods to enhance detection, public health investigations, and surveillance to address this global threat.

Gonococcal Conjunctivitis in Adults: Case Report and Retrospective Review of Cases in Alberta, Canada, 2000-2016.

BACKGROUND: A case of gonococcal conjunctivitis (GC) prompted us to review the reported cases and treatment regimens of GC in Alberta, Canada. METHODS: Gonococcal cases reported from 2000 to 2016 were extracted from the provincial sexually transmitted infection reporting database. The diagnosis of GC was based on a positive culture and/or nucleic acid amplification test from the eye. RESULTS: A total of 45 cases of GC were reported in adults. Three quarters (75.6%; n = 34) of the cases were diagnosed using culture, 57.8% (n = 26) of cases were among men, and 55.5% (n = 25) were diagnosed since 2014. Very few (13.3%; n = 6) of the cases were treated according to current Canadian Guidelines on Sexually Transmitted Infections, using 2 g of ceftriaxone in combination with azithromycin or doxycycline. Results of test of cures were available for 15.6% (n = 7) of the cases and occurred within 10 to 79 days (median = 26 days) after treatment; all were negative. CONCLUSIONS: Gonococcal conjunctivitis was relatively uncommon in our region, but given its potential for severe manifestations and sequelae coupled with the rising rates of gonorrhea; it remains important to consider this diagnosis in sexually active individuals presenting with purulent conjunctivitis. Additional studies are needed to inform treatment recommendations and to evaluate outcomes of infection.

Gonococcal and Chlamydial Cases of Pelvic Inflammatory Disease at 2 Canadian Sexually Transmitted Infection Clinics, 2004 to 2014: A Retrospective Cross-sectional Review.

One hundred thirteen patients with gonococcal and chlamydial pelvic inflammatory disease were reviewed at 2 Canadian sexually transmitted infection clinics. Most patients (81%) with pelvic inflammatory disease were diagnosed as having chlamydia alone. Three treatment failures were seen in patients treated with ofloxacin.

Retrospective Review of Gonococcal and Chlamydial Cases of Epididymitis at 2 Canadian Sexually Transmitted Infection Clinics, 2004-2014.

Fifty-seven cases of gonococcal and chlamydial infections complicated by acute epididymitis seen at 2 Alberta STI clinics from 2004 to 2014 were reviewed. The majority responded to treatment recommended by national guidelines. Three of 6 treatment failures were not treated according to guidelines.

Trichomonas vaginalis Prevalence and Correlates in Women and Men Attending STI Clinics in Western Canada.

Trichomonas vaginalis prevalence (2.8%) in female sexually transmitted infection clinic attendees was within the prevalence of chlamydia (5.8%) and gonorrhea (1.8%), while being very low for male attendees (0.2%). Correlates among women were indigenous ethnicity, other ethnicity, and being symptomatic.

A Retrospective Review of Treatment Failures Using Azithromycin and Doxycycline in the Treatment of Rectal Chlamydia Infections in Women and Men Who Have Sex With Men.

We examined the prevalence of rectal chlamydia treatment failures in men who have sex with men and women attending Alberta sexually transmitted infection clinics. Among those completing a test of cure, there was no significant difference among patients treated initially with azithromycin (treatment failure, 39/460 [8.5%]; 95% confidence interval, 5.9%-11.0%) compared with patients treated with doxycycline (0/16; 95% confidence interval, 0%-0.2%; P = 0.63).

Impact of Introducing Triage Criteria for Express Testing at a Canadian Sexually Transmitted Infection Clinic.

The implementation of express testing in an sexually transmitted infection (STI) clinic reduced the length of visit time compared with other visit types and increased the proportion of STIs diagnosed at clinic visits. Express testing did not impact the time to treatment for asymptomatic patients diagnosed as having an STI.

Sensitivity and specificity of two investigational Point of care tests for Syphilis and HIV (PoSH Study) for the diagnosis and treatment of infectious syphilis in Canada: a cross-sectional study.

OBJECTIVES: Single-visit testing and treatment for syphilis can reduce follow-up visits. The objectives of this study were to evaluate the performance and treatment outcomes of two dual syphilis/HIV point-of-care tests (POCTs). METHODS: Participants aged 16 years and older were offered concurrent syphilis/HIV POCTs with fingerstick blood sampling using two extremely rapid (<5 minutes) devices (MedMira Multiplo Rapid TP/HIV test and INSTI Multiplex HIV-1/HIV-2/Syphilis Antibody Test). Those with positive POCT results were offered same-day syphilis treatment and linkage to HIV care. Nurses performed testing at two emergency departments, a First Nations community, a correctional facility, and a sexually transmitted infection clinic. POCT results were compared with those of standard serological testing. Sensitivity and specificity were calculated. RESULTS: Between August 2020 and February 2022, 1526 visits were completed. Both POCTs accurately identified participants with HIV (sensitivity, 100% [24 of 24]; 95% CI, 86.2-100%; specificity, 99.6% [1319 of 1324]; 95% CI, 99.1-99.8%), linking 24 HIV cases to care. Both tests were most sensitive with a rapid plasma reagin (RPR) of ≥1:8 dilutions (Multiplo: sensitivity, 98.3% [231 of 235]; 95% CI, 95.7-99.3%; specificity, 99.5% [871 of 875]; 95% CI, 98.8-99.8%; INSTI Multiplex: sensitivity, 97.9% [230 of 235]; 95% CI, 95.1-99.1%; specificity, 99.8% [873 of 875]; 95% CI, 99.2-99.9%) and least sensitive with non-reactive RPR (Multiplo: sensitivity, 54.1% [59 of 109]; 95% CI, 44.8-63.2%; specificity, 99.5% [871 of 875]; 95% CI, 98.8-99.8%; INSTI Multiplex: sensitivity, 28.4% [31 of 109]; 95% CI, 20.8-37.5%; specificity, 99.8% [873 of 875]; 95% CI, 99.2-99.9%). Eighty-five percent of participants with infectious syphilis were treated on the same day as the positive POCT result. DISCUSSION: Two extremely rapid (<5 minutes) dual syphilis/HIV POCTs showed excellent sensitivity and specificity for the diagnosis of active syphilis (RPR, ≥1:8 dilutions) and HIV and confirmed the ability to offer single-visit testing and treatment for syphilis and linkage to HIV care in diverse clinical settings.

Retrospective testing for the presence of monkeypox virus in a high-risk population from February-June 2022 in Alberta, Canada.

Background: A multi-country outbreak of monkeypox virus (MPXV) infections was identified by the World Health Organization in May 2022. The western Canadian province of Alberta identified its first case of MPXV in a returning traveller on June 2, 2022. We undertook a retrospective testing exercise to evaluate whether MPXV may have been circulating in the province earlier. Methods: Skin (genital and non-genital) and mucosal lesion swabs submitted for herpes simplex virus (HSV)/varicella zoster virus (VZV)/syphilis testing from male patients attending sexually-transmitted infection clinics across the province of Alberta from January 28 to May 30, 2022 were retrieved from storage. The population tested was selected based on the epidemiology of the current 2022 multi-country MPXV outbreak. Samples underwent viral nucleic acid extraction and testing for the presence of Orthopoxvirus DNA using a commercial real-time polymerase chain reaction (PCR) kit. Results: A total of 392 samples (representing 341 unique individuals of median age 31 years) were retrieved. Of them, 349 (89.0%) samples were submitted for HSV/VZV/syphilis testing, 13 (3.3%) for HSV/VZV only, and 30 (7.7%) for syphilis PCR only. None of the 392 samples tested were found to be positive for Orthopoxvirus DNA. Conclusions: The results of this study indicate that circulation of MPXV in a higher-risk population in Alberta, prior to the first case, was less likely. We recommend that other provinces/territories review their local epidemiology, context and resources prior to conducting similar studies.

Historique: En mai 2022, l'Organisation mondiale de la Santé a déclaré une flambée multinationale d'infection par le virus de la variole simienne (MPXV). Le 2 juin 2022, la province de l'Alberta, dans l'Ouest canadien, a recensé son premier cas de MPXV chez un voyageur de retour de l'étranger. Les chercheurs ont entrepris un exercice de dépistage rétrospectif pour évaluer la possibilité que le MPXV ait circulé auparavant dans la province. Méthodologie: Les chercheurs ont extrait de l'entreposage les écouvillons des lésions cutanées (génitales et non génitales) et muqueuses soumis en vue de dépister le virus herpès simplex (VHS), le virus varicelle-zona (VZV) et le virus de la syphilis des patients de sexe masculin qui avaient fréquenté les cliniques d'infections transmises sexuellement de la province de l'Alberta entre le 28 janvier et le 30 mai 2022. Ils ont sélectionné la population soumise au dépistage en fonction de l'épidémiologie de la flambée multinationale de MPXV en 2022. Les écouvillons ont été soumis à l'extraction et au test des acides nucléiques viraux pour dépister la présence d'ADN de l'Orthopoxvirus au moyen d'un test commercial d'amplification en chaîne par polymérase (PCR). RÉsultats: Les chercheurs ont extrait un total de 392 échantillons (représentant 341 personnes uniques d'un âge médian de 31 ans). De ce nombre, 349 (89,0 %) avaient été soumis au test PCR du VHS, du VZV et de la syphilis, 13 (3,3 %), du VHS et du VZV seulement et 30 (7,7 %), de la syphilis seulement. Aucun des 392 échantillons n'a donné de résultat positif à l'ADN de l'Orthopoxvirus. Conclusions: D'après les résultats de la présente étude, il est peu probable que le MPXV ait circulé dans la population plus vulnérable de l'Alberta avant la détection du premier cas. Les chercheurs recommandent que les autres provinces et territoires examinent leur épidémiologie locale, le contexte et les ressources avant de procéder à des études de ce type.

Outcomes of infectious syphilis in pregnant patients and maternal factors associated with congenital syphilis diagnosis, Alberta, 2017-2020.

Background: Congenital syphilis (CS) is a significant public health challenge, requiring early diagnosis and treatment to improve infant outcomes. The aim of this study is to describe public health outcomes of infectious syphilis cases among pregnant patients and factors associated with a CS diagnosis for their infant. Methods: We conducted a retrospective review of demographic and clinical characteristics of infectious syphilis cases diagnosed during pregnancy and resulting infant outcomes in Alberta from 2017 to 2020 from the provincial communicable disease database. Adequate maternal treatment was defined as receiving at least one dose of Benzathine penicillin G-LA 2.4 million units IM at least 28 days before delivery. Univariate and multivariate analysis was performed to determine factors associated with CS diagnosis using SPSS version 25. Results: A total of 374 cases of infectious syphilis were diagnosed in pregnancy, with two patients being diagnosed twice in a single pregnancy. The majority (79.1%; n=296) of women had a live birth, followed by therapeutic abortion (9.4%; n=35), stillbirth (7.5%; n=28) and spontaneous abortion (4.0%; n=15). Infant records (n=265) were available for review (n=117 CS cases and 148 non-cases). Correlates associated with CS were screening time in third trimester (adjusted odds ratio [AOR] 8.4, 95% confidence interval [CI], 2.9-24.6) and fewer than 28 days before delivery (AOR 8.1, 1.4-47.8 [vs. first and second trimester] and inadequate treatment (AOR 86.1, CI, 15.9-466.5). Among the CS cases, 23.1% (n=27) were stillborn compared with one (0.7%) stillbirth in the non-CS infants (p<0.001). Conclusion: The early identification and treatment of syphilis in pregnancy is crucial to preventing poor infant outcomes.