ABSTRACT
Current guideline recommends the use of two identification methods for Neisseria gonorrhoeae. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) is now used for primary identification and may be sufficient for definitive identification of N. gonorrhoeae. The performance of three secondary tests (BactiCard, RapID NH and NET test) were compared using 45 bacterial isolates, including 37 Neisseria species. These secondary tests demonstrated diminished specificity (67% - 88%) for N. gonorrhoeae compared with MALDI-TOF. Additionally, data from six clinical microbiology laboratories was used to compare confirmatory test costs and the agreement of results with MALDI-TOF. Discrepancies were documented for 9.4% of isolates, though all isolates (n= 288) identified by MALDI-TOF as N. gonorrhoeae were confirmed by the reference laboratory. These data demonstrate that MALDI-TOF alone is sufficient for N. gonorrhoeae identification, as secondary did not add diagnostic value but do add costs to the testing process.
Subject(s)
Gonorrhea , Neisseria gonorrhoeae , Sensitivity and Specificity , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Neisseria gonorrhoeae/isolation & purification , Neisseria gonorrhoeae/classification , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/economics , Humans , Gonorrhea/diagnosis , Gonorrhea/microbiology , Bacteriological Techniques/economics , Bacteriological Techniques/methodsABSTRACT
OBJECTIVES: International infectious disease/obstetrical societies have recently recommended universal hepatitis C virus (HCV) prenatal screening and these same recommendations are forthcoming in Canada. At present, there is no formal analysis of universal HCV screening or linkage to care of pregnant people in Ontario. The objectives of our study were to determine the seroprevalence of HCV using 2 different methods to evaluate universal screening, as well as identify opportunities that may improve linkage to care. METHODS: To assess seroprevalence in a large urban area, we aimed to test 12 000 de-identified samples submitted for prenatal HIV testing in the catchment area of Toronto Public Health for HCV antibodies. Then, to assess the seroprevalence as well as the operational impact and follow-up in a real-world setting, we completed a Quality Improvement Project (QIP) for 1 year at a large tertiary care obstetrical centre in London, Ontario. RESULTS: From 2019 to 2021, 11 999 de-identified samples were screened from Toronto with a seroprevalence of 0.40 (95% CI 0.29-0.53). In London, 5771 people were screened in 2021 with a seroprevalence of 0.55% (95% CI 0.38-0.78). Taken together, those aged 26-35 years had the highest positivity; in the QIP, 9% had no documented risk factor, and 59% of individuals were not linked to the next step in HCV care. CONCLUSIONS: HCV prenatal seroprevalence in Ontario is comparable to hepatitis B virus, and â¼15-30-fold higher than HIV. Diagnosis in pregnancy is critical to facilitate referrals for treatment between pregnancies and could increase screening among children born to positive women.
Subject(s)
Hepatitis C , Mass Screening , Pregnancy Complications, Infectious , Humans , Female , Ontario/epidemiology , Hepatitis C/epidemiology , Hepatitis C/diagnosis , Pregnancy , Seroepidemiologic Studies , Pregnancy Complications, Infectious/epidemiology , Pregnancy Complications, Infectious/diagnosis , Adult , Mass Screening/methods , Prevalence , Prenatal Diagnosis/methods , Prenatal CareABSTRACT
We evaluated the performance of the Copan Transystem™ M40 collection swab containing Amies agar gel ('M40') on the Aptima® TV Assay for the detection of Trichomonas vaginalis ribosomal RNA (rRNA) using a novel 'Single Swab' molecular workflow. Overall agreement between Aptima TV Assay and wet mount microscopy was 99 % (152/153; 95 % confidence interval 0.9806 to 1.006), a positive agreement of 100 % and negative agreement of 99 %. Limit of detection for microscopy was 100,000-fold higher compared to molecular. T. vaginalis rRNA was stable in M40 under room-temperature and refrigerated conditions. The 'Single Swab' workflow resulted in a 57.4 % reduction in hands-on time, and a 5-fold increase in technologist productivity. Post-molecular test implementation analysis demonstrated a 2.27-fold increase in T. vaginalis positivity rate compared to the pre-implementation method. Collectively, our 'Single Swab' molecular testing approach was non-inferior to wet mount microscopy with the added benefits of a simplified and more efficient workflow.
Subject(s)
Galanin , Peptide Fragments , Trichomonas Vaginitis , Trichomonas vaginalis , Humans , Female , Trichomonas vaginalis/genetics , Trichomonas Vaginitis/diagnosis , Sensitivity and Specificity , AgarABSTRACT
The GENCOV study aims to identify patient factors which affect COVID-19 severity and outcomes. Here, we aimed to evaluate patient characteristics, acute symptoms and their persistence, and associations with hospitalization. Participants were recruited at hospital sites across the Greater Toronto Area in Ontario, Canada. Patient-reported demographics, medical history, and COVID-19 symptoms and complications were collected through an intake survey. Regression analyses were performed to identify associations with outcomes including hospitalization and COVID-19 symptoms. In total, 966 responses were obtained from 1106 eligible participants (87% response rate) between November 2020 and May 2022. Increasing continuous age (aOR: 1.05 [95%CI: 1.01-1.08]) and BMI (aOR: 1.17 [95%CI: 1.10-1.24]), non-White/European ethnicity (aOR: 2.72 [95%CI: 1.22-6.05]), hypertension (aOR: 2.78 [95%CI: 1.22-6.34]), and infection by viral variants (aOR: 5.43 [95%CI: 1.45-20.34]) were identified as risk factors for hospitalization. Several symptoms including shortness of breath and fever were found to be more common among inpatients and tended to persist for longer durations following acute illness. Sex, age, ethnicity, BMI, vaccination status, viral strain, and underlying health conditions were associated with developing and having persistent symptoms. By improving our understanding of risk factors for severe COVID-19, our findings may guide COVID-19 patient management strategies by enabling more efficient clinical decision making.
Subject(s)
COVID-19 , Humans , COVID-19/epidemiology , Hospitalization , Inpatients , Ontario/epidemiology , Risk FactorsABSTRACT
Rapid advancements of genome sequencing (GS) technologies have enhanced our understanding of the relationship between genes and human disease. To incorporate genomic information into the practice of medicine, new processes for the analysis, reporting, and communication of GS data are needed. Blood samples were collected from adults with a PCR-confirmed SARS-CoV-2 (COVID-19) diagnosis (target N = 1500). GS was performed. Data were filtered and analyzed using custom pipelines and gene panels. We developed unique patient-facing materials, including an online intake survey, group counseling presentation, and consultation letters in addition to a comprehensive GS report. The final report includes results generated from GS data: (1) monogenic disease risks; (2) carrier status; (3) pharmacogenomic variants; (4) polygenic risk scores for common conditions; (5) HLA genotype; (6) genetic ancestry; (7) blood group; and, (8) COVID-19 viral lineage. Participants complete pre-test genetic counseling and confirm preferences for secondary findings before receiving results. Counseling and referrals are initiated for clinically significant findings. We developed a genetic counseling, reporting, and return of results framework that integrates GS information across multiple areas of human health, presenting possibilities for the clinical application of comprehensive GS data in healthy individuals.
Subject(s)
COVID-19 , Genetic Counseling , Adult , Humans , COVID-19/epidemiology , COVID-19/genetics , SARS-CoV-2/genetics , Genomics/methods , GenotypeABSTRACT
BACKGROUND: Widespread testing for severe acute respiratory coronavirus virus 2 (SARS-CoV-2) is necessary to curb the spread of coronavirus disease 2019 (COVID-19), but testing is undermined when the only option is a nasopharyngeal swab. Self-collected swab techniques can overcome many of the disadvantages of a nasopharyngeal swab, but they require evaluation. METHODS: Three self-collected non-nasopharyngeal swab techniques (saline gargle, oral swab and combined oral-anterior nasal swab) were compared to a nasopharyngeal swab for SARS-CoV-2 detection at multiple COVID-19 assessment centers in Toronto, Canada. The performance characteristics of each test were assessed. RESULTS: The adjusted sensitivity of the saline gargle was 0.90 (95% CI 0.86-0.94), the oral swab was 0.82 (95% CI, 0.72-0.89) and the combined oral-anterior nasal swab was 0.87 (95% CI, 0.77-0.93) compared to a nasopharyngeal swab, which demonstrated a sensitivity of Ë90% when all positive tests were the reference standard. The median cycle threshold values for the SARS-CoV-2 E-gene for concordant and discordant saline gargle specimens were 17 and 31 (P < .001), for the oral swabs these values were 17 and 28 (P < .001), and for oral-anterior nasal swabs these values were 18 and 31 (P = .007). CONCLUSIONS: Self-collected saline gargle and an oral-anterior nasal swab have a similar sensitivity to a nasopharyngeal swab for the detection of SARS-CoV-2. These alternative collection techniques are cheap and can eliminate barriers to testing, particularly in underserved populations.
Subject(s)
COVID-19 , Outpatients , Humans , Nasopharynx , SARS-CoV-2 , Saliva , Specimen HandlingABSTRACT
Widely available and easily accessible testing for COVID-19 is a cornerstone of pandemic containment strategies. Nasopharyngeal swabs (NPS) are the currently accepted standard for sample collection but are limited by their need for collection devices and sampling by trained healthcare professionals. The aim of this study was to compare the performance of saliva to NPS in an outpatient setting. This was a prospective study conducted at three centers, which compared the performance of saliva and NPS samples collected at the time of assessment center visit. Samples were tested by real-time reverse transcription polymerase chain reaction and sensitivity and overall agreement determined between saliva and NPS. Clinical data was abstracted by chart review for select study participants. Of the 432 paired samples, 46 were positive for SARS-CoV-2, with seven discordant observed between the two sample types (four individuals testing positive only by NPS and three by saliva only). The observed agreement was 98.4% (kappa coefficient 0.91) and a composite reference standard demonstrated sensitivity of 0.91 and 0.93 for saliva and NPS samples, respectively. On average, the Ct values obtained from saliva as compared to NPS were higher by 2.76. This study demonstrates that saliva performs comparably to NPS for the detection of SARS-CoV-2. Saliva was simple to collect, did not require transport media, and could be tested with equipment readily available at most laboratories. The use of saliva as an acceptable alternative to NPS could support the use of widespread surveillance testing for SARS-CoV-2.
Subject(s)
COVID-19 Nucleic Acid Testing , COVID-19/diagnosis , Nasopharynx/virology , Outpatients/statistics & numerical data , SARS-CoV-2/isolation & purification , Saliva/virology , Adult , Female , Humans , Limit of Detection , Male , Middle Aged , Ontario , Prospective Studies , RNA, Viral/genetics , Sensitivity and Specificity , Specimen HandlingABSTRACT
BACKGROUND: Ontario is 1 of 5 provinces that immunize adolescents for hepatitis B virus (HBV), despite the World Health Organization recommendation for universal birth dose vaccination. One rationale for not vaccinating at birth is that universal prenatal screening and related interventions prevent vertical transmission. The aims of our study were to evaluate the uptake and epidemiology of prenatal HBV screening, and to determine the number of children in Ontario with a diagnosis of HBV before adolescent vaccination. METHODS: We extracted data from ICES, Public Health Ontario and Better Outcomes & Registry Network (BORN) Ontario databases. We assessed prenatal screening uptake and prevalence of prenatal hepatitis B surface antigen (HBsAg) from 2012 to 2016, as well as subsequent hepatitis B e-antigen (HBeAg) and HBV DNA testing and percent positivity. We used age and region to subcategorize the results. In a separate unlinked analysis, we evaluated the number of children positive for HBV aged 0-11 years who were born in Ontario from 2003 to 2013. RESULTS: From 2012 to 2016, 93% of pregnant women were screened for HBV, with an HBsAg prevalence of 0.6%. Prevalence of HBsAg increased with age, peaking at older than 45 years at 3%. North Toronto had the highest overall prevalence of 1.5%, whereas northern Ontario had the lowest. Of women who were HBsAg positive, HBeAg and HBV DNA tests were subsequently ordered in 13% and 38%, respectively. Of children born in Ontario between 2003 and 2013, 139 of 23 759 tested positive for HBV. INTERPRETATION: Prenatal HBV screening is not universal and subsequent evaluation is poor, limiting optimal intervention and possibly contributing to some Ontario-born children being given a diagnosis of HBV before age 12 years. These findings underscore the limitations of the province's adolescent vaccination strategy.
Subject(s)
Hepatitis B/epidemiology , Infectious Disease Transmission, Vertical/prevention & control , Pregnancy Complications, Infectious/epidemiology , Prenatal Diagnosis , Adolescent , Adult , Age Factors , Child , Child Health Services , Child, Preschool , Cost of Illness , Female , Hepatitis B/prevention & control , Hepatitis B Vaccines , Humans , Infant , Infant, Newborn , Male , Middle Aged , Ontario/epidemiology , Pregnancy , Pregnancy Complications, Infectious/prevention & control , Prevalence , Registries , Young AdultABSTRACT
Zika virus (ZIKV) is a mosquito-borne flavivirus associated with a febrile illness as well as severe complications, including microcephaly and Guillain-Barré Syndrome. Antibody cross-reactivity between flaviviruses has been documented, and in regions where ZIKV is circulating, dengue virus (DENV) is also endemic, leaving the potential that previous exposure to DENV could alter clinical features of ZIKV infection. To investigate this, we performed a retrospective case-control study in which we compared Canadian travellers who had been infected with ZIKV and had serological findings indicating previous DENV or other flavivirus exposure (n = 16) to those without any previous exposure (n = 44). Patient samples were collected between February 2016 and September 2017 and submitted to Public Health Ontario for testing. ZIKV infection was determined using real-time RT-PCR and antibodies against DENV were identified by the plaque-reduction neutralization test. The mean time from symptom onset to sample collection was 5 days for both groups; the magnitude of viremia was not statistically different (Ct values: 35.6 vs. 34.9, p-value = 0.2). Clinical scores were also similar. Our findings indicate that previous DENV or other flavivirus exposure did not result in greater viremia or a higher illness score.
Subject(s)
Antibodies, Viral/blood , Dengue Virus/immunology , Viremia , Zika Virus Infection/immunology , Zika Virus Infection/virology , Zika Virus/isolation & purification , Adult , Canada , Case-Control Studies , Humans , Middle Aged , Real-Time Polymerase Chain Reaction , Retrospective Studies , Reverse Transcriptase Polymerase Chain Reaction , Severity of Illness Index , Travel-Related IllnessABSTRACT
The administration of antibiotics while critical for treatment, can be accompanied by potentially severe complications. These include toxicities associated with the drugs themselves, the selection of resistant organisms and depletion of endogenous host microbiota. In addition, antibiotics may be associated with less well-recognized complications arising through changes in the pathogens themselves. Growing evidence suggests that organisms exposed to antibiotics can respond by altering the expression of toxins, invasins and adhesins, as well as biofilm, resistance and persistence factors. The clinical significance of these changes continues to be explored; however, it is possible that treatment with antibiotics may inadvertently precipitate a worsening of the clinical course of disease. Efforts are needed to adjust or augment antibiotic therapy to prevent the transition of pathogens to hypervirulent states. Better understanding the role of antibiotic-microbe interactions and how these can influence disease course is critical given the implications on prescription guidelines and antimicrobial stewardship policies.
ABSTRACT
In order to expand hepatitis C virus (HCV) screening, a change in the diagnostic paradigm is warranted to improve accessibility and decrease costs, such as utilizing dried blood spot (DBS) collection. In our study, blood from 68 patients with chronic HCV infection was spotted onto DBS cards and stored at the following temperatures for one week: -80 °C, 4 °C, 21 °C, 37 °C, and alternating 37 °C and 4 °C; to assess whether temperature change during transportation would affect sensitivity. Sample was eluted from the DBS cards and tested for HCV antibodies (HCV-Ab) and HCV core antigen (core-Ag). HCV-Abs were detected from 68/68 DBS samples at -80 °C, 4 °C, 21 °C, and 67/68 at 37 °C and alternating 37 °C and 4 °C. Sensitivity of core-Ag was as follows: 94% (-80 °C), 94% (4 °C), 91% (21 °C), 93% (37 °C), and 93% (37 °C/4 °C). Not only did temperature not greatly affect sensitivity, but sensitivities are higher than previously reported, and support the use of this assay as an alternative to HCV RNA. We then completed a head-to-head comparison (n = 49) of venous versus capillary samples, and one versus two DBS. No difference in core-Ag sensitivity was observed by sample type, but there was an improvement when using two spots. We conclude that HCV-Abs and core-Ag testing from DBS cards has high diagnostic accuracy and could be considered as an alternative to HCV RNA in certain settings.
Subject(s)
Dried Blood Spot Testing/methods , Hepacivirus/immunology , Hepatitis C Antigens/analysis , Hepatitis C/blood , Adult , Aged , Female , Hepacivirus/genetics , Hepacivirus/isolation & purification , Hepatitis C/diagnosis , Hepatitis C/virology , Hepatitis C Antibodies/analysis , Hepatitis C Antibodies/immunology , Hepatitis C Antigens/immunology , Humans , Male , Middle AgedABSTRACT
The isolation of stool pathogens is difficult due to their fastidious nature and the rapid overgrowth of fecal flora. In this study, we evaluate the preservation and isolation of enteric pathogens from stool using the automated COPAN Walk-Away Specimen Processor (WASP®) in conjunction with FecalSwab™ and selenite media. Pathogen viability and fecal commensal abundance were stable in FecalSwab™ media under both room-temperature and refrigerated incubation conditions, resulting in a significantly increased number of well-isolated pathogen colonies observed when compared to samples incubated in modified Cary-Blair media. Isolation of individual pathogen colonies was improved via WASP® planting compared to those planted using the Isoplater system. Furthermore, preincubation using the newly formulated COPAN selenite media significantly enhanced the yield of Salmonella enterica serovar Typhimurium. Together, the automated WASP® system combined with FecalSwab™ and selenite media represents a rapid and efficient approach for the processing of stool specimens compared to standard methods.
Subject(s)
Automation, Laboratory/methods , Bacteria/isolation & purification , Bacterial Infections/diagnosis , Bacteriological Techniques/methods , Feces/microbiology , Gastroenteritis/diagnosis , Specimen Handling/methods , Culture Media/chemistry , TemperatureABSTRACT
Carbapenemase producing Enterobacteriaceae (CPE) are becoming a global healthcare concern. Current laboratory methods for the detection of CPE include screening followed by confirmatory phenotypic and genotypic tests. These processes would generally take ≥72 hours, which could negatively impact patient care and Infection Control practices. To this end, we developed a protocol for rapid resistance testing (RRT) to detect hydrolysis in a panel of beta lactam antibiotics consisting of ampicillin, cefazolin, cefotaxime and imipenem, using liquid chromatography tandem mass spectrometry. Ninety-nine beta lactamase producing Enterobacteriaceae isolates were used to evaluate the RRT method, 54 isolates were CPE and 45 isolates were Class A or AmpC beta lactamase producing Enterobacteriaceae but not carbapenemase producers. We also tested 10 E.coli isolates that were susceptible to ampicillin, cefazolin, cefotaxime and imipenem. Receiver Operating Characteristic (ROC) Curves analysis showed that imipenem had a sensitivity and a specificity of 100% for crabapenemase detection at hydrolysis cut off values that are greater than 50% and less than or equal to 80%. The RRT protocol can be conducted in a time frame of less than 2 hours. This preliminary study shows that the rapid resistance testing protocol might have utility for the rapid detection of CPE. Additional work with a greater number and variety of beta- lactamase producing Enterobacteriaceae isolates is required to validate these preliminary findings.
Subject(s)
Anti-Bacterial Agents/pharmacology , Bacterial Proteins/isolation & purification , Carbapenem-Resistant Enterobacteriaceae/metabolism , Microbial Sensitivity Tests/methods , Tandem Mass Spectrometry/methods , beta-Lactam Resistance , beta-Lactamases/isolation & purification , Anti-Bacterial Agents/metabolism , Anti-Bacterial Agents/therapeutic use , Bacterial Proteins/metabolism , Carbapenem-Resistant Enterobacteriaceae/drug effects , Carbapenem-Resistant Enterobacteriaceae/isolation & purification , Carbapenems/metabolism , Carbapenems/pharmacology , Carbapenems/therapeutic use , Cephalosporins/metabolism , Cephalosporins/pharmacology , Cephalosporins/therapeutic use , Chromatography, High Pressure Liquid/instrumentation , Chromatography, High Pressure Liquid/methods , Enterobacteriaceae Infections/drug therapy , Enterobacteriaceae Infections/microbiology , Escherichia coli/drug effects , Escherichia coli/metabolism , Humans , Hydrolysis , Microbial Sensitivity Tests/instrumentation , Tandem Mass Spectrometry/instrumentation , Time Factors , beta-Lactamases/metabolismABSTRACT
We evaluated a direct from positive blood culture pelleting procedure that utilizes a lysis-centrifugation protocol for the identification of microorganisms by MALDI-TOF MS with subsequent antimicrobial susceptibility testing (AST) and rapid methicillin- and beta-lactam-resistance screening. The identification evaluation was performed on 125 cultures and resulted in the correct genus-level identification in 91.2% of cultures and a species-level concordance of 82.4% compared to routine subcultured growth. For the AST evaluation, susceptibility results from direct pelleting and subcultured growth for 187 cultures were compared; an average ±2-fold dilution agreement of 98.2% (1650/1681) and 98.6% (1375/1394) for Gram-negatives and -positives, respectively, was found. Major errors fell below 5% except for MRSA, which was falsely reported as oxacillin sensitive 17.2% (11/66) of the time. Lastly, the sensitivity and specificity of rapid MRSA screening were 94.7% (36/38) and 90.0% (9/10), respectively, while the ESBL screening results were 90.3% (65/72) and 100.0% (13/13) respectively.
Subject(s)
Blood Culture , Microbial Sensitivity Tests , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Anti-Bacterial Agents/pharmacology , Bacteremia/diagnosis , Bacteremia/microbiology , Bacteria/classification , Bacteria/drug effects , Bacteria/isolation & purification , Bacteriological Techniques , Blood Culture/methods , Centrifugation/methods , Humans , Sensitivity and Specificity , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methodsABSTRACT
The Zika virus (ZIKV) epidemic is an ongoing public health concern. ZIKV is a flavivirus reported to be associated with microcephaly, and recent work in animal models demonstrates the ability of the virus to cross the placenta and affect fetal brain development. Recent findings suggest that the virus preferentially infects neural stem cells and thereby deregulates gene expression, cell cycle progression, and increases cell death. However, neuronal stem cells are not the only brain cells that are susceptible to ZIKV and infection of other brain cells may contribute to disease progression. Herein, we characterized ZIKV replication in astrocytes, and profiled temporal changes in host microRNAs (miRNAs) and transcriptomes during infection. We observed the deregulation of numerous processes known to be involved in flavivirus infection, including genes involved in the unfolded protein response pathway. Moreover, a number of miRNAs were upregulated, including miR-30e-3p, miR-30e-5p, and, miR-17-5p, which have been associated with other flavivirus infections. This study highlights potential miRNAs that may be of importance in ZIKV pathogenesis.
Subject(s)
Astrocytes/metabolism , Astrocytes/virology , MicroRNAs/genetics , RNA, Messenger/genetics , Zika Virus/pathogenicity , Animals , Astrocytes/pathology , Cell Line , Female , Gene Expression , Humans , Microarray Analysis , Pregnancy , Up-Regulation , Virus Replication , Zika Virus/physiologyABSTRACT
With the emerging Zika virus (ZIKV) epidemic, serologic diagnosis relies on a labor-intensive IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) and confirmation by a plaque reduction neutralization test (PRNT). To streamline serologic testing, several commercial assays have been developed. Our aim was to compare the commercial Euroimmun anti-ZIKV IgM and IgG assays to the reference MAC-ELISA and PRNT currently in use. Serum specimens submitted to Public Health Ontario Laboratory, Canada, were tested for IgM and IgG using the Euroimmun assays and the results were compared with those from MAC-ELISA. The PRNT was performed on positive or equivocal specimens using either MAC-ELISA or Euroimmun assays, MAC-ELISA-inconclusive specimens, and a convenience sample of specimens negative by both assays (cohort 1). Another set of specimens selected on the basis of PRNT results was subsequently tested by the Euroimmun assays (cohort 2). MAC-ELISA was positive, equivocal, negative, and inconclusive in 57/223, 15/223, 147/223, and 4/223 specimens, respectively. Among the 76 specimens that were MAC-ELISA positive, equivocal, or inconclusive, 30 (39.5%) were Euroimmun IgM and/or IgG positive or equivocal. Among the 147 MAC-ELISA-negative specimens, 136 (92.5%) were Euroimmun IgM and IgG negative. The sensitivity of the combined Euroimmun IgM/IgG against the PRNT was 83% (cohort 1) and 92% (cohort 2), whereas the specificity was 81% (cohort 1) and 65% (cohort 2). The combined Euroimmun IgM/IgG showed good specificity (92.5%) but suboptimal sensitivity (39.5%) compared with that of the MAC-ELISA. However, the sensitivity of the combined Euroimmun IgM/IgG against the PRNT was significantly higher (83 to 92%). More studies are needed before commercial assays are implemented for routine ZIKV serologic diagnosis.
Subject(s)
Antibodies, Viral/blood , Enzyme-Linked Immunosorbent Assay/methods , Immunoglobulin G/blood , Immunoglobulin M/blood , Serologic Tests/methods , Zika Virus Infection/diagnosis , Zika Virus/immunology , Adult , Cohort Studies , Female , Humans , Male , Middle Aged , Ontario , Pregnancy , Sensitivity and SpecificityABSTRACT
UNLABELLED: The capacity of subinhibitory levels of antibiotics to modulate bacterial virulence in vitro has recently been brought to light, raising concerns over the appropriateness of low-dose therapies, including antibiotic prophylaxis for recurrent urinary tract infection management. However, the mechanisms involved and their relevance in influencing pathogenesis have not been investigated. We characterized the ability of antibiotics to modulate virulence in the uropathogens Staphylococcus saprophyticus and Escherichia coli. Several antibiotics were able to induce the expression of adhesins critical to urothelial colonization, resulting in increased biofilm formation, colonization of murine bladders and kidneys, and promotion of intracellular niche formation. Mice receiving subinhibitory ciprofloxacin treatment were also more susceptible to severe infections and frequent recurrences. A ciprofloxacin prophylaxis model revealed this strategy to be ineffective in reducing recurrences and worsened infection by creating larger intracellular reservoirs at higher frequencies. Our study indicates that certain agents used for antibiotic prophylaxis have the potential to complicate infections. IMPORTANCE: Antibiotics are the mainstay treatment for bacterial infections; however, evidence is emerging that argues these agents may have off-target effects if sublethal concentrations are present. Most studies have focused on changes occurring in vitro, leaving questions regarding the clinical relevance in vivo. We utilized a murine urinary tract infection model to explore the potential impact of low-dose antibiotics on pathogenesis. Using this model, we showed that subinhibitory antibiotics prime uropathogens for adherence and invasion of murine urothelial tissues. These changes in initial colonization promoted the establishment of chronic infection. Furthermore, treatment of chronically infected mice with subtherapeutic ciprofloxacin served to exacerbate infection. A part of these changes was thought to be due to suppression of mucosal immunity, as demonstrated through reductions in cytokine secretion and migration of leukocytes into the urinary tract. This work identifies novel risk factors associated with antibiotic therapy when dosing strategies fall below subtherapeutic levels.
Subject(s)
Anti-Bacterial Agents/administration & dosage , Bacterial Infections/drug therapy , Escherichia coli/drug effects , Escherichia coli/growth & development , Staphylococcus saprophyticus/drug effects , Staphylococcus saprophyticus/growth & development , Urinary Tract Infections/drug therapy , Adhesins, Bacterial/metabolism , Animals , Bacterial Adhesion , Bacterial Infections/immunology , Biofilms/growth & development , Disease Models, Animal , Drug Therapy/methods , Escherichia coli/immunology , Escherichia coli/physiology , Female , Kidney/microbiology , Mice, Inbred C3H , Mice, Inbred C57BL , Recurrence , Staphylococcus saprophyticus/immunology , Staphylococcus saprophyticus/physiology , Urinary Bladder/microbiology , Urinary Tract Infections/immunology , VirulenceABSTRACT
INTRODUCTION: The aim of this study was to examine endogenous biotin levels in tumour specimens collected from patients with renal and testicular tumours and compare them to the surrounding non-neoplastic surgical margin. METHODS: Frozen samples were obtained from the Ontario Tumour Bank. Renal and testicular tumour tissue were included in this study. Normal tissue from the negative surgical margins of each tumour served as a control. Biotin detection in tissue specimens was determined using immunohistochemistry (IHC). RESULTS: Specimens collected from 56 patients (36 men and 20 women) were included in this study. Histopathology of the 52 renal tumours included 31 (60%) conventional type RCC, 5 (10%) chromophobe RCC, 5 (10%) papillary RCC, 1 (2%) oncocytoma and 10 (19%) upper tract urothelial carcinoma (UC). The 4 testicular tumours included 1 seminomatous (25%) germ cell tumour and 3 (75%) non seminomatous germ cell tumours. CONCLUSION: No biotin signal was perceived in all tested tumour samples. Endogenous biotin expression was detected in the matching non-neoplastic surgical margin of tested renal tissues. This lack of staining may prove to be a valuable tool in future studies.
ABSTRACT
INTRODUCTION: Ochratoxin-A (OTA) is one of the most abundant food-contaminating mycotoxins, known for its nephrotoxicity, neurotoxicity, gonadotoxicity, teratogenicity, immunosuppression and carcinogenesis. OTA has been linked to several genitourinary pathologies, including Balkan nephropathy and genitourinary malignancies. We examine OTA levels in serum samples and tumour specimens collected from patients with renal and testicular tumours. METHODS: Frozen samples were obtained from the Ontario Tumour Bank. Serum specimens, along with renal and testicular tumour biopsies, were included in this study. Normal tissue from the negative surgical margins of each tumour served as a control. OTA levels in serum was measured using the enzyme-linked immunosorbent assay (ELISA), while OTA detection in tissue specimens was determined using immunohistochemistry (IHC). RESULTS: We included specimens collected from 56 patients (36 men and 20 women). Histopathology of the 52 renal tumours included 31 (60%) conventional type renal cell carcinomas (RCC), 5 (10%) chromophobe RCC, 5 (10%) papillary RCC, 1 (2%) oncocytoma and 10 (19%) upper tract urothelial carcinoma (UC). The 4 testicular tumours included 1 seminomatous (25%) germ cell tumour and 3 (75%) non-seminomatous germ cell tumours. OTA was detected in the serum of renal tumour patients, with a range from 0.004 to 0.25 ng/mL (mean: 0.07 and median 0.06 ng/mL). There was no OTA signal detected by IHC staining in all tested renal and testicular tumours. CONCLUSIONS: The OTA levels detected in the serum of patients were highly variable and relatively low. No OTA was detected in the tissue samples.