Diagnosis of Helicobacter pylori infection: serology vs. urea breath test.

The objective of the study was to ascertain an optimal Helicobacter pylori diagnostic strategy using population-level laboratory data comparing the performance of serology against urea breath test (UBT). H. pylori diagnostic test results for serology and UBT from two laboratories over a 12-year period (2006-20017) were extracted, linked, and analyzed. A subset of this population underwent both methods of testing within days of each other, enabling a direct comparison of the two methods. The average prevalence of H. pylori positivity was 21.3% by serology and 17.5% by UBT. There were 2,612 individuals who had serology performed first, followed by UBT within 14 days. For this subset, the sensitivity of serology compared with UBT was 96.5% with a specificity of 79.2%. The negative predictive value for serology was 98.4%. Contrary to various recent clinical guidelines, the data show that serology still has utility as a sensitive enough test to be used as an initial H. pylori screening test in a lower prevalence population. Negative serology can be used with confidence to rule out active infection, whereas a positive serology could be followed up with a UBT or a similar performing test such as stool antigen to differentiate active from past infection. For population-based diagnostic recommendations, such a strategy may be ideal since serology generally costs less than UBT and may be combined with a blood draw being done for other diagnostic tests. Continuing to offer serology increases options for patients and may provide economic benefits for single-payer health care systems or health maintenance organizations. IMPORTANCE: This study compares the performance of serology with urea breath test in the diagnosis of Helicobacter pylori in a population-level data set and mimics a head-to-head direct comparison as the study population had both tests performed within 2 weeks of each other. This provides new information supporting the use of serology in a diagnostic algorithm. There are several instances where serology could be preferable to patients to rule out disease, despite some guidelines suggesting serology should not be used.

Mapping Ixodes pacificus and Borrelia burgdorferi Habitat Suitability Under Current and Mid-Century Climate in the Pacific Northwest (BC and WA).

Introduction: Lyme disease is the most common vector-borne disease in the United States and Canada. The primary vector for the causative agent of Lyme disease, Borrelia burgdorferi, in the Pacific Northwest is the western blacklegged tick, Ixodes pacificus. Materials and Methods: Using active tick surveillance data from British Columbia, Canada, and Washington State, USA, habitat suitability models using MaxEnt (maximum entropy) were developed for I. pacificus to predict its current and mid-century geographic distributions. Passive surveillance data both from BC and WA were also visualized. Results: According to the constructed models, the number of frost-free days during the winter is the most relevant predictor of its habitat suitability, followed by summer climate moisture, ecoregion, and mean minimum fall temperature. The ensemble geographic distribution map predicts that the coastal regions and inland valleys of British Columbia and the Puget Lowlands of Washington State provide the most suitable habitats for I. pacificus. The density map of ticks submitted from passive surveillance data was overlaid on the current distribution map and demonstrates the correlation between numbers of submissions and habitat suitability. Mid-century projections, based on current climate change predictions, indicate a range expansion, especially of low and moderate suitability, from current distribution. Regarding Lyme disease risk, I. pacificus identified from both active and passive surveillance and tested positive for B. burgdorferi were found to be in areas of moderate to very high suitability for I. pacificus. Conclusion: According to developed models, the total suitable habitat area for I. pacificus will expand in the interior regions of British Columbia and Washington State. However, the risk remains small given relatively low infection rates among I. pacificus. Further studies are required to better understand how this might change in the future.

Whole-genome sequencing of Western Canadian Borrelia spp. collected from diverse tick and animal hosts reveals short-lived local genotypes interspersed with longer-lived continental genotypes.

Changing climates are allowing the geographic expansion of ticks and their animal hosts, increasing the risk of Borrelia-caused zoonoses in Canada. However, little is known about the genomic diversity of Borrelia from the west of the Canadian Rockies and from the tick vectors Ixodes pacificus, Ixodes auritulus and Ixodes angustus. Here, we report the whole-genome shotgun sequences of 51 Borrelia isolates from multiple tick species collected on a range of animal hosts between 1993 and 2016, located primarily in coastal British Columbia. The bacterial isolates represented three different species from the Lyme disease-causing Borrelia burgdorferi sensu lato genospecies complex [Borrelia burgdorferi sensu stricto (n=47), Borrelia americana (n=3) and Borrelia bissettiae (n=1)]. The traditional eight-gene multi-locus sequence typing (MLST) strategy was applied to facilitate comparisons across studies. This identified 13 known Borrelia sequence types (STs), established 6 new STs, and assigned 5 novel types to the nearest sequence types. B. burgdorferi s. s. isolates were further differentiated into ten ospC types, plus one novel ospC with less than 92 % nucleotide identity to all previously defined ospC types. The MLST types resampled over extended time periods belonged to previously described STs that are distributed across North America. The most geographically widespread ST, ST.12, was isolated from all three tick species. Conversely, new B. burgdorferi s. s. STs from Vancouver Island and the Vancouver region were only detected for short periods, revealing a surprising transience in space, time and host tick species, possibly due to displacement by longer-lived genotypes that expanded across North America.This article contains data hosted by Microreact.

Lyme Disease Confirmatory Western Blot Is Redundant for Screen Negative Samples in Low Endemic Areas, British Columbia, Canada.

Background: Borrelia burgdorferi sensu stricto is the causative agent of Lyme disease (LD). Possible early symptoms include flu-like symptoms and erythema migrans and later, the risk of disruption of the nervous system, joints, and heart. A two-tiered testing method is employed for serological diagnostics. The Public Health Agency of Canada guidelines recommend that samples tested negative on first-tiered test need not be confirmed by second-tiered test. Due to the challenging nature of diagnosis leading to misconceptions among physicians about false negatives, confirmatory testing is requested despite the initial negative result. Methods: Hundred screen-negative Lyme patient samples from 2007 to 2016 were tested by Western blot (WB) second-tiered confirmatory test upon physician's request in British Columbia to study the first-tiered screening test sufficiency. Results: Those negative for first-tiered enzyme-linked immunosorbent assay were also negative by WB. Conclusion: Results demonstrate that confirmatory testing is not necessary on screen-negative samples. Hence, first-tiered test is sufficient to rule out LD.

Passive tick surveillance and detection of Borrelia burgdorferi in ticks from companion animals in British Columbia: 2018 to 2020.

Objective: The present study was designed to identify tick species and determine prevalence of Borrelia burgdorferi infection in ticks obtained from companion animals in British Columbia. Animals and samples: Ticks were submitted by British Columbia veterinarians from client-owned companion animals over a 31-month period. Procedure: Each tick was identified and PCR testing for B. burgdorferi undertaken on all Ixodes species identified by the Zoonotic Diseases and Emerging Pathogens Section of British Columbia Centre for Disease Control Public Health Laboratory (BCCDC PHL). Results: Overall, 85% (n = 300) of ticks submitted were Ixodes spp., with the majority known to transmit B. burgdorferi. Furthermore, 0.8% (95% confidence interval: 0.094 to 2.78%) of these ticks were PCR-positive for B. burgdorferi. Conclusion and clinical relevance: Although the B. burgdorferi positivity rate in this study was low, it remains important for veterinary professionals to inform pet owners that ticks are present and can pose a risk to pets and humans. In eastern North America, B. burgdorferi infection risk has increased rapidly, underscoring the importance of ongoing surveillance in British Columbia to understand current and future distributions of ticks and tick-borne pathogens, especially in the context of climate change.

Surveillance passive des tiques et détection de Borrelia burgdorferi chez des tiques provenant d'animaux de compagnie en Colombie-Britannique: 2018 à 2020. Objectif: Cette étude a été élaboré afin d'identifier les espèces de tiques et de déterminer la prévalence de l'infection à Borrelia burgdorferi chez des tiques obtenues d'animaux de compagnie en Colombie-Britannique. Animaux et échantillons: Les tiques ont été soumises par des médecins vétérinaires de la Colombie-Britannique obtenues d'animaux de compagnie de clients sur une période de 31 mois. Procédure: Chaque tique a été identifiée et un test PCR pour détecter B. burdorferi réalisé sur toutes les espèces Ixodes identifiées par la Section des maladies zoonotiques et des agents pathogènes émergents du Centre for Disease Control Public Health Laboratory de la Colombie-Britannique. Résultats: Au total, 85 % (n = 300) des tiques soumises étaient des Ixodes spp., dont la majorité reconnue pour transmettre B. burgdorferi. De plus, 0,8 % (intervalle de confiance 95 %: 0,094 à 2,78 %) de ces tiques étaient positives pour B. burgdorferi par PCR. Conclusion et signification clinique: Bien que le taux de positivité pour B. burgdorferi dans la présente étude soit faible, il n'en demeure pas moins important pour les professionnels vétérinaires d'informer les propriétaires d'animaux de compagnie que les tiques sont présentes et peuvent représenter un risque pour les animaux de compagnie et les humains. Dans le nord de l'Amérique du Nord, le risque d'infection par B. burgdorferi a augmenté rapidement, soulignant l'importance d'une surveillance continue en Colombie-Britannique pour comprendre la distribution actuelle et future des tiques et agents pathogènes transmis par les tiques, spécialement dans le contexte des changements climatiques.(Traduit par Dr Serge Messier).

Case Series of Jamestown Canyon Virus Infections with Neurologic Outcomes, Canada, 2011-2016.

Jamestown Canyon virus (JCV) is a mosquitoborne orthobunyavirus in the California serogroup that circulates throughout Canada and the United States. Most JCV exposures result in asymptomatic infection or a mild febrile illness, but JCV can also cause neurologic diseases, such as meningitis and encephalitis. We describe a case series of confirmed JCV-mediated neuroinvasive disease among persons from the provinces of British Columbia, Alberta, Quebec, and Nova Scotia, Canada, during 2011-2016. We highlight the case definitions, epidemiology, unique features and clinical manifestations, disease seasonality, and outcomes for those cases. Two of the patients (from Quebec and Nova Scotia) might have acquired JCV infections during travel to the northeastern region of the United States. This case series collectively demonstrates JCV's wide distribution and indicates the need for increased awareness of JCV as the underlying cause of meningitis/meningoencephalitis during mosquito season.

Adaptive immune responses to two-dose COVID-19 vaccine series in healthy Canadian adults ≥ 50 years: a prospective, observational cohort study.

To evaluate immune responses to COVID-19 vaccines in adults aged 50 years and older, spike protein (S)-specific antibody concentration, avidity, and function (via angiotensin-converting enzyme 2 (ACE2) inhibition surrogate neutralization and antibody dependent cellular phagocytosis (ADCP)), as well as S-specific T cells were quantified via activation induced marker (AIM) assay in response to two-dose series. Eighty-four adults were vaccinated with either: mRNA/mRNA (mRNA-1273 and/or BNT162b2); ChAdOx1-S/mRNA; or ChAdOx1-S/ChAdOx1-S. Anti-S IgG concentrations, ADCP scores and ACE2 inhibiting antibody concentrations were highest at one-month post-second dose and declined by four-months post-second dose for all groups. mRNA/mRNA and ChAdOx1-S/mRNA schedules had significantly higher antibody responses than ChAdOx1-S/ChAdOx1-S. CD8+ T-cell responses one-month post-second dose were associated with increased ACE2 surrogate neutralization. Antibody avidity (total relative avidity index) did not change between one-month and four-months post-second dose and did not significantly differ between groups by four-months post-second dose. In determining COVID-19 correlates of protection, a measure that considers both antibody concentration and avidity should be considered.

Factors associated with SARS-CoV-2 infection in unvaccinated children and young adults.

BACKGROUND AND OBJECTIVES: Pediatric COVID-19 cases are often mild or asymptomatic, which has complicated estimations of disease burden using existing testing practices. We aimed to determine the age-specific population seropositivity and risk factors of SARS-CoV-2 seropositivity among children and young adults during the pandemic in British Columbia (BC). METHODS: We conducted two cross-sectional serosurveys: phase 1 enrolled children and adults < 25 years between November 2020-May 2021 and phase 2 enrolled children < 10 years between June 2021-May 2022 in BC. Participants completed electronic surveys and self-collected finger-prick dried blood spot (DBS) samples. Samples were tested for immunoglobulin G antibodies against ancestral spike protein (S). Descriptive statistics from survey data were reported and two multivariable analyses were conducted to evaluate factors associated with seropositivity. RESULTS: A total of 2864 participants were enrolled, of which 95/2167 (4.4%) participants were S-seropositive in phase 1 across all ages, and 61/697 (8.8%) unvaccinated children aged under ten years were S-seropositive in phase 2. Overall, South Asian participants had a higher seropositivity than other ethnicities (13.5% vs. 5.2%). Of 156 seropositive participants in both phases, 120 had no prior positive SARS-CoV-2 test. Young infants and young adults had the highest reported seropositivity rates (7.0% and 7.2% respectively vs. 3.0-5.6% across other age groups). CONCLUSIONS: SARS-CoV-2 seropositivity among unvaccinated children and young adults was low in May 2022, and South Asians were disproportionately infected. This work demonstrates the need for improved diagnostics and reporting strategies that account for age-specific differences in pandemic dynamics and acceptability of testing mechanisms.

Draft whole-genome sequences of three Borrelia burgdorferi isolates from Western Canada.

Whole-genome sequences are presented for three Borrelia burgdorferi, a causative agent of Lyme disease in North America, isolated from Ixodes pacificus ticks collected in British Columbia, Canada. Shotgun DNA libraries were prepared with Illumina DNA Prep and sequenced using the MiniSeq platform. Genome assemblies enabled multilocus sequence typing and ospC typing.

Uptake of SARS-CoV-2 vaccination among structurally-marginalized people who use drugs in Vancouver, Canada.

We sought to evaluate the rates and predictors of SARS-CoV-2 vaccination among members of a structurally-marginalized population of people who use drugs (PWUD) during a targeted, community-wide, vaccination campaign in Vancouver, Canada. Interviewer-administered data were collected from study participants between June 2021 and March 2022. Generalized estimating equation analysis was used to identify factors associated with SARS-CoV-2 vaccine uptake, ascertained through a province-wide vaccine registry. Among 223 PWUD, 107 (48.0%) reported receipt of at least two SARS-CoV-2 vaccine doses at baseline and this increased to 151 (67.7%) by the end of the study period. Using social media as a source of vaccine information was negatively associated with SARS-CoV-2 vaccine uptake (Adjusted odds ratio [AOR] 0.27, 95% confidence interval [CI] 0.09-0.81) and HIV seropositivity (AOR 2.68, 95% CI 1.12-6.39) and older age (AOR 1.27, 95% CI 1.07-1.51) were positively associated with SARS-CoV-2 vaccine uptake. These findings suggest that the targeted vaccination campaign in Vancouver may be an effective model to promote SARS-CoV-2 vaccination in other jurisdictions. However, using social media as a source of vaccine information likely reduced SARS-CoV-2 vaccine uptake in PWUD arguing for further efforts to promote accessible and evidence-based vaccine information among marginalized populations.

Plasmodium knowlesi Infection in Traveler Returning to Canada from the Philippines, 2023.

A 55-year-old man sought treatment for an uncomplicated febrile illness after returning to Canada from the Philippines. A suspected diagnosis of Plasmodium knowlesi infection was confirmed by PCR, and treatment with atovaquone/proguanil brought successful recovery. We review the evolving epidemiology of P. knowlesi malaria in the Philippines, specifically within Palawan Island.

Examining the impacts of a syphilis awareness campaign among gay, bisexual, and other men who have sex with men (gbMSM) in British Columbia, Canada.

OBJECTIVES: Syphilis rates have increased in BC and disproportionately affect gay, bisexual, and other men who have sex with men (gbMSM). A social marketing campaign (Syphistory) ran from January to September 2017 with the primary goal of increasing syphilis knowledge and a secondary goal of increasing syphilis screening among gbMSM in BC. METHODS: We used pre- and post-campaign surveys to assess changes in syphilis knowledge from a convenience sample of clients attending STI clinics using one-sided t-tests. We used online Piwik metrics to examine the campaign reach, and provincial testing data to examine trends in syphilis screening. We used data from the Engage Study to examine factors associated with campaign awareness and associations with syphilis testing. RESULTS: Of the 2155 visitors to the Syphistory website with known geography, 79.4% were from BC. Moreover, STI clinic participants who saw the campaign demonstrated a greater knowledge of syphilis (9.7/12, 80.8%) than those who did not see the campaign (mean 8.9/12, 74%) (p < 0.001). Provincial syphilis testing rates were 8764 and 9749 in the 12 months before and after the campaign; however, we did not find an overall trend in testing before versus after the campaign (p = 0.147). Among Engage participants, 12.7% reported seeing the campaign and we found an association between campaign exposure and recent syphilis testing (aOR = 2.73; 95% CI = 1.51, 4.93). CONCLUSION: gbMSM who saw the campaign were more likely to report being tested for syphilis in the previous 6 months. STI clinic attendees who reported seeing the campaign also had higher syphilis knowledge compared to those who did not.

RéSUMé: OBJECTIFS: Les taux de syphilis ont augmenté en Colombie-Britannique et affectent de manière disproportionnée les hommes gais, bisexuels et autres hommes ayant des relations sexuelles avec des hommes (gbHARSAH). Une campagne de marketing social (Syphistory) a été mené de janvier à septembre 2017 avec pour objectif principal d'informer sur la syphilis et pour objectif secondaire d'augmenter le dépistage de la syphilis chez les gbHARSAH en Colombie-Britannique. MéTHODES: Nous avons réalisé deux sondages, l'un avant et l'autre après la campagne, sur un échantillon de convenance constitué de patients fréquentant des cliniques ITS, pour évaluer les changements dans les connaissances sur la syphilis à l'aide de tests t unilatéraux. Nous avons utilisé les mesures Piwik en ligne pour examiner la portée de la campagne et les données provinciales sur les tests pour examiner les tendances quant au dépistage de la syphilis. Nous avons utilisé les données de l'étude Engage à Vancouver, pour identifier les facteurs associés à la sensibilisation lors de la campagne et les associations avec le dépistage de la syphilis. RéSULTATS: Sur les 2 155 visiteurs du site Web Syphistory dont la position géographique était connue, 79,4 % provenaient de la Colombie-Britannique. De plus, les participants aux cliniques ITS ayant vu la campagne ont démontré une meilleure connaissance de la syphilis (9,7/12, 80,8 %) par rapport à ceux n'ayant pas vu la campagne (moyenne 8,9/12, 74 %) (p<0,001). Les taux provinciaux de dépistage de la syphilis étaient de 8 764 et 9 749 au cours des 12 mois précédant et suivant la campagne; cependant, nous n'avons pas trouvé de tendance globale à la hausse des dépistages suite à la campagne (p=0,147). Parmi les participants Engage, 12,7 % ont déclaré avoir vu la campagne en ligne et nous avons trouvé une association entre l'exposition à la campagne et le dépistage récent de la syphilis (RCa=2,73; IC à 95 %=1,51, 4,93). CONCLUSION: Les gbHARSAH qui ont vu la campagne étaient plus susceptibles de déclarer avoir été testés pour la syphilis au cours des six derniers mois. Les participants aux cliniques ITS qui ont déclaré avoir vu la campagne avaient également une meilleure connaissance de la syphilis que ceux qui ne l'ont pas vue.

Surveillance for Ixodes scapularis and Ixodes pacificus ticks and their associated pathogens in Canada, 2020.

Background: Ixodes scapularis and Ixodes pacificus ticks are the principal vectors of the agent of Lyme disease and several other tick-borne diseases in Canada. Tick surveillance data can be used to identify local tick-borne disease risk areas and direct public health interventions. The objective of this article is to describe the seasonal and spatial characteristics of the main Lyme disease vectors in Canada, and the tick-borne pathogens they carry, using passive and active surveillance data from 2020. Methods: Passive and active surveillance data were compiled from the National Microbiology Laboratory Branch (Public Health Agency of Canada), provincial and local public health authorities, and eTick (an online, image-based platform). Seasonal and spatial analyses of ticks and their associated pathogens are presented, including infection prevalence estimates. Results: In passive surveillance, I. scapularis (n=7,534) were submitted from all provinces except Manitoba and British Columbia, while I. pacificus (n=718) were submitted only from British Columbia. No ticks were submitted from the Territories. The seasonal distribution of I. scapularis submissions was bimodal, but unimodal for I. pacificus. Four tick-borne pathogens were identified in I. scapularis (Borrelia burgdorferi, Anaplasma phagocytophilum, Babesia microti and Borrelia miyamotoi) and one in I. pacificus (B. miyamotoi). In active surveillance, I. scapularis (n=688) were collected in Ontario, Québec and New Brunswick. Five tick-borne pathogens were identified: B. burgdorferi, A. phagocytophilum, B. microti, B. miyamotoi and Powassan virus. Conclusion: This article provides a snapshot of the distribution of I. scapularis and I. pacificus and their associated human pathogens in Canada in 2020, which can help assess the risk of exposure to tick-borne pathogens in different provinces.

Successful treatment of suspected early form of chronic Chagas cardiomyopathy: a case report.

Background: Chagas disease, caused by the protozoan Trypanosoma cruzi, is the most common parasitic aetiology of non-ischaemic cardiomyopathy in the Americas, causing significant morbidity and mortality. The clinical spectrum ranges from early asymptomatic disease to severe cardiac manifestations including dilated cardiomyopathy, heart failure, dysrhythmias, conduction abnormalities, thromboembolism, and sudden death. Case summary: We present a case of Chagas disease in a 75-year-old patient originally from El Salvador who presented to our Canadian tertiary centre with heart failure and atrial fibrillation/flutter. The patient had dilated cardiomyopathy with severely reduced systolic function, which was thought to be early Chagas cardiomyopathy after confirmatory positive serologies for T. cruzi. The patient demonstrated significant clinical improvement and recovery of systolic function with benznidazole therapy that was sustained up to 12 months on follow up. Discussion: The American Heart Association recommends considering treatment of early chronic Chagas cardiomyopathy with anti-trypanosomal therapy. Our case highlights the importance of multidisciplinary collaboration in the diagnosis of early Chagas cardiomyopathy and critical timing of benznidazole, as effectiveness is limited in late disease due to myocardial cell-death programme. Although the historical BENEFIT study is known to not have shown mortality reduction, we advocate that the significant reduction in cardiovascular-related hospitalizations should be considered for symptomatic patients with early Chagas cardiomyopathy with the potential benefit of improving cardiac function and avoiding need for heart transplantation.

Characterizing Longitudinal Antibody Responses in Recovered Individuals Following COVID-19 Infection and Single-Dose Vaccination: A Prospective Cohort Study.

BACKGROUND: Investigating antibody titers in individuals who have been both naturally infected with SARS-CoV-2 and vaccinated can provide insight into antibody dynamics and correlates of protection over time. METHODS: Human coronavirus (HCoV) IgG antibodies were measured longitudinally in a prospective cohort of qPCR-confirmed, COVID-19 recovered individuals (k = 57) in British Columbia pre- and post-vaccination. SARS-CoV-2 and endemic HCoV antibodies were measured in serum collected between Nov. 2020 and Sept. 2021 (n = 341). Primary analysis used a linear mixed-effects model to understand the effect of single dose vaccination on antibody concentrations adjusting for biological sex, age, time from infection and vaccination. Secondary analysis investigated the cumulative incidence of high SARS-CoV-2 anti-spike IgG seroreactivity equal to or greater than 5.5 log10 AU/mL up to 105 days post-vaccination. No re-infections were detected in vaccinated participants, post-vaccination by qPCR performed on self-collected nasopharyngeal specimens. RESULTS: Bivariate analysis (complete data for 42 participants, 270 samples over 472 days) found SARS-CoV-2 spike and RBD antibodies increased 14-56 days post-vaccination (p < 0.001) and vaccination prevented waning (regression coefficient, B = 1.66 [95%CI: 1.45-3.46]); while decline of nucleocapsid antibodies over time was observed (regression coefficient, B = -0.24 [95%CI: -1.2-(-0.12)]). A positive association was found between COVID-19 vaccination and endemic human ß-coronavirus IgG titer 14-56 days post vaccination (OC43, p = 0.02 & HKU1, p = 0.02). On average, SARS-CoV-2 anti-spike IgG concentration increased in participants who received one vaccine dose by 2.06 log10 AU/mL (95%CI: 1.45-3.46) adjusting for age, biological sex, and time since infection. Cumulative incidence of high SARS-CoV-2 spike antibodies (>5.5 log10 AU/mL) was 83% greater in vaccinated compared to unvaccinated individuals. CONCLUSIONS: Our study confirms that vaccination post-SARS-CoV-2 infection provides multiple benefits, such as increasing anti-spike IgG titers and preventing decay up to 85 days post-vaccination.

A diagnostic classifier for gene expression-based identification of early Lyme disease.

Background: Lyme disease is a tick-borne illness that causes an estimated 476,000 infections annually in the United States. New diagnostic tests are urgently needed, as existing antibody-based assays lack sufficient sensitivity and specificity. Methods: Here we perform transcriptome profiling by RNA sequencing (RNA-Seq), targeted RNA-Seq, and/or machine learning-based classification of 263 peripheral blood mononuclear cell samples from 218 subjects, including 94 early Lyme disease patients, 48 uninfected control subjects, and 57 patients with other infections (influenza, bacteremia, or tuberculosis). Differentially expressed genes among the 25,278 in the reference database are selected based on ≥1.5-fold change, ≤0.05 p value, and ≤0.001 false-discovery rate cutoffs. After gene selection using a k-nearest neighbor algorithm, the comparative performance of ten different classifier models is evaluated using machine learning. Results: We identify a 31-gene Lyme disease classifier (LDC) panel that can discriminate between early Lyme patients and controls, with 23 genes (74.2%) that have previously been described in association with clinical investigations of Lyme disease patients or in vitro cell culture and rodent studies of Borrelia burgdorferi infection. Evaluation of the LDC using an independent test set of samples from 63 subjects yields an overall sensitivity of 90.0%, specificity of 100%, and accuracy of 95.2%. The LDC test is positive in 85.7% of seronegative patients and found to persist for ≥3 weeks in 9 of 12 (75%) patients. Conclusions: These results highlight the potential clinical utility of a gene expression classifier for diagnosis of early Lyme disease, including in patients negative by conventional serologic testing.

British Columbia's Experience after Implementation of the Treponema pallidum Reverse Algorithm and PCR Detection, 2015 to 2020.

British Columbia (BC) implemented the syphilis reverse screening algorithm and Treponema pallidum PCR testing in 2014. We summarize the performance characteristics of the algorithm, together with PCR direct detection, and report on syphilis cases identified from 2015 to 2020. Prior to 2015, samples for syphilis diagnosis were first screened by rapid plasma reagin (RPR). As of 2015, sera were screened by the Siemens Advia Centaur syphilis assay (enzyme immunoassay [EIA]). Positive and equivocal samples were reflex tested by a T. pallidum passive particle agglutination assay (TPPA) and RPR. We used T. pallidum DNA PCR on clinical samples and restriction fragment length polymorphism analysis to identify azithromycin resistance mutations. Case/epidemiological data were obtained from the BC surveillance system. Of 1,631,519 samples screened by the EIA, 72,492 (4.4%) were positive and 187 (<0.1%) were equivocal. Of EIA-positive/equivocal samples, 10.6% were false positive, and false positivity was higher at lower EIA indices. The reverse algorithm detected 4,693 late latent syphilis cases that likely would have been missed by RPR screening. PCR had a very high sensitivity of 100% versus 52.9% and 52.4% for dark-field (DF) and immunofluorescence (IF) microscopy, respectively. The azithromycin resistance mutation A2058G was identified in 96% of PCR-positive samples, and A2059G was identified in 4%. Annually, there were 944 to 1,467 syphilis cases, with 62% in men who reported male sexual partners. The reverse algorithm had a low false-positive rate and very few equivocal screening results but did identify previously undiagnosed late latent syphilis cases. PCR was more sensitive than both DF and IF microscopy for direct diagnosis and enabled monitoring for azithromycin resistance. IMPORTANCE In this study, we summarize the performance characteristics of the algorithm, together with PCR direct detection and epidemiological analysis, and report on syphilis cases identified from 2015 to 2020. This allowed us to paint a complete picture of the outcome of the utilization of the reverse algorithm for diagnosing syphilis cases. The study clearly showed that the reverse algorithm had a low false-positive rate and very few equivocal screening results but did identify previously undiagnosed late latent syphilis cases. PCR was more sensitive than both DF and IF microscopy for direct diagnosis and enabled monitoring for azithromycin resistance.