RESUMEN
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.
Asunto(s)
Virus de la Encefalitis de California , Encefalitis de California , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Canadá/epidemiología , Virus de la Encefalitis de California/genética , Encefalitis de California/epidemiología , Encefalitis de California/virología , Historia del Siglo XXIRESUMEN
Inherited forms of arrhythmogenic and dilated cardiomyopathy (ACM and DCM) are characterized by variable disease expression and age-related penetrance. Calcium (Ca2+) is crucially important for proper cardiac function, and dysregulation of Ca2+ homeostasis seems to underly cardiomyopathy etiology. A polymorphism, c.286T>G p.(Ser96Ala), in the gene encoding the histidine-rich Ca2+ binding (HRC) protein, relevant for sarcoplasmic reticulum Ca2+ cycling, has previously been associated with a marked increased risk of life-threatening arrhythmias among idiopathic DCM patients. Following this finding, we investigated whether p.(Ser96Ala) affects major cardiac disease manifestations in carriers of the phospholamban (PLN) c.40_42delAGA; p.(Arg14del) pathogenic variant (cohort 1); patients diagnosed with, or predisposed to, ACM (cohort 2); and DCM patients (cohort 3). We found that the allele frequency of the p.(Ser96Ala) polymorphism was similar across the general European-American population (control cohort, 40.3-42.2%) and the different cardiomyopathy cohorts (cohorts 1-3, 40.9-43.9%). Furthermore, the p.(Ser96Ala) polymorphism was not associated with life-threatening arrhythmias or heart failure-related events across various patient cohorts. We therefore conclude that there is a lack of evidence supporting the important role of the HRC p.(Ser96Ala) polymorphism as a modifier in cardiomyopathy, refuting previous findings. Further research is required to identify bona fide genomic predictors for the stratification of cardiomyopathy patients and their risk for life-threatening outcomes.
Asunto(s)
Cardiomiopatías , Cardiomiopatía Dilatada , Humanos , Arritmias Cardíacas/genética , Arritmias Cardíacas/metabolismo , Calcio/metabolismo , Proteínas de Unión al Calcio/genética , Proteínas de Unión al Calcio/metabolismo , Cardiomiopatías/genética , Cardiomiopatía Dilatada/genética , Histidina/genética , Polimorfismo GenéticoRESUMEN
The aim of presenting this case was to show the difficulties in making the differential diagnosis of viral meningitis during the COVID pandemic situation.
Asunto(s)
COVID-19 , Herpes Simple , Herpesvirus Humano 1 , Meningitis Aséptica , Diagnóstico Diferencial , Herpes Simple/diagnóstico , Humanos , Meningitis Aséptica/diagnósticoRESUMEN
The evaluation of humoral protective immunity against SARS-CoV-2 remains crucial in understanding both natural immunity and protective immunity conferred by the several vaccines implemented in the fight against COVID-19. The reference standard for the quantification of antibodies capable of neutralizing SARS-CoV-2 is the plaque-reduction neutralization test (PRNT). However, given that it is a laboratory-developed assay, validation is crucial in order to ensure sufficient specificity and intra- and interassay precision. In addition, a multitude of other serological assays have been developed, including enzyme-linked immunosorbent assay (ELISA), flow cytometry-based assays, luciferase-based lentiviral pseudotype assays, and commercially available human ACE2 receptor-blocking antibody tests, which offer practical advantages in the evaluation of the protective humoral response against SARS-CoV-2. In this study, we validated a SARS-CoV-2 PRNT to assess both 50% and 90% neutralization of SARS-CoV-2 according to guidelines outlined by the World Health Organization. Upon validation, the reference-standard PRNT demonstrated excellent specificity and both intra- and interassay precision. Using the validated assay as a reference standard, we characterized the neutralizing antibody response in specimens from patients with laboratory-confirmed COVID-19. Finally, we conducted a small-scale multilaboratory comparison of alternate SARS-CoV-2 PRNTs and surrogate neutralization tests. These assays demonstrated substantial to perfect interrater agreement with the reference-standard PRNT and offer useful alternatives to assess humoral immunity against SARS-CoV-2. IMPORTANCE SARS-CoV-2, the causal agent of COVID-19, has infected over 246 million people and led to over 5 million deaths as of October 2021. With the approval of several efficacious COVID-19 vaccines, methods to evaluate protective immune responses will be crucial for the understanding of long-term immunity in the rapidly growing vaccinated population. The PRNT, which quantifies SARS-CoV-2-neutralizing antibodies, is used widely as a reference standard to validate new platforms but has not undergone substantial validation to ensure excellent inter- and intraassay precision and specificity. Our work is significant, as it describes the thorough validation of a PRNT, which we then used as a reference standard for the comparison of several alternative serological methods to measure SARS-CoV-2-neutralizing antibodies. These assays demonstrated excellent agreement with the reference-standard PRNT and include high-throughput platforms, which can greatly enhance capacity to assess both natural and vaccine-induced protective immunity against SARS-CoV-2.
Asunto(s)
Prueba Serológica para COVID-19/métodos , COVID-19/diagnóstico , COVID-19/inmunología , Inmunidad Humoral/inmunología , Pruebas de Neutralización/métodos , SARS-CoV-2/inmunología , Enzima Convertidora de Angiotensina 2 , Animales , Anticuerpos Neutralizantes , Anticuerpos Antivirales/inmunología , Vacunas contra la COVID-19/inmunología , Chlorocebus aethiops , Pruebas Diagnósticas de Rutina , Ensayo de Inmunoadsorción Enzimática/métodos , Células HEK293 , Humanos , Inmunidad , SARS-CoV-2/aislamiento & purificación , Sensibilidad y Especificidad , Células VeroRESUMEN
There remains an urgent need for assays to quantify humoral protective immunity to SARS-CoV-2 to understand the immune responses of COVID-19 patients, evaluate efficacy of vaccine candidates in clinical trials, and conduct large-scale epidemiological studies. The plaque-reduction neutralization test (PRNT) is the reference-standard for quantifying antibodies capable of neutralizing SARS-CoV-2. However, the PRNT is logistically demanding, time-consuming, and requires containment level-3 facilities to safely work with live virus. In contrast, a surrogate virus neutralization test (sVNT) manufactured by Genscript is a quick and simple assay that detects antibodies that inhibit the RBD-ACE2 interaction, crucial for virus entry into host cells. In this study, we evaluate the sensitivity, specificity, and cross-reactivity of the sVNT compared with the PRNT using both 50% and 90% SARS-CoV-2 neutralization as a reference-standard. We found that the sVNT provides a high-throughput screening tool prior to confirmatory PRNT testing for the evaluation of SARS-CoV-2 neutralizing antibodies.
Asunto(s)
Anticuerpos Neutralizantes/sangre , Anticuerpos Antivirales/sangre , SARS-CoV-2/inmunología , Ensayo de Placa Viral/métodos , Enzima Convertidora de Angiotensina 2/metabolismo , COVID-19/diagnóstico , Ensayos Analíticos de Alto Rendimiento/métodos , Humanos , Pruebas de Neutralización/métodosRESUMEN
Background: Jamestown Canyon virus (JCV) and snowshoe hare virus (SSHV) are wide-ranging mosquito-borne arboviruses in the California serogroup viruses (CSGV) that are known to circulate in New Brunswick. Despite potential for debilitating central nervous system manifestations, the prevalence of human exposure to these viruses in New Brunswick is unknown. The goal of this study was to quantify rates of human exposure in New Brunswick to these neglected arboviruses. Methods: A retrospective, anonymized provincial serosurvey was performed using a stratified random sample of residual sera submitted between May 2015 and August 2016. To determine the seroprevalence of JCV and SSHV, competitive enzyme-linked immunosorbent assay-positive samples were confirmed positive using plaque-reduction neutralization testing (PRNT). Results: A total of 452 serum samples were screened. The seroprevalence of antibodies against CSGV was estimated to be 31.6% (95% CI 27.4% to 36.1%) with 143 positive samples. PRNT results indicated that most single virus exposures were due to JCV (38 of 143; 26.6%) rather than SSHV (3 of 143; 2.1%). The species of CSGV, to which the remaining 102 seropositive people were exposed, could not be precisely determined. Conclusions: The prevalence of human exposure to CSGV is high but comparable to rates observed in other Atlantic Canadian jurisdictions. Studies such as this provide important baseline epidemiological data regarding the risk of exposure to these neglected arboviruses. SSHV and JCV should be considered in the differential diagnosis for undiagnosed febrile and neuroinvasive illness during mosquito season, particularly when testing for common aetiologies is negative or inconclusive.
Historique : Le virus de Jamestown Canyon (VJC) et le virus du lièvre d'Amérique (VLA) sont des arbovirus à grande portée transmis par des moustiques des virus du sérogroupe Californie (VSGC) qui circulent au Nouveau-Brunswick (NB). Malgré le risque de manifestations débilitantes du système nerveux central, on ne connaît pas la prévalence d'exposition humaine à ces virus au NB. La présente étude visait à quantifier le taux d'exposition humaine à ces arbovirus négligés au NB. Méthodologie : Les chercheurs ont réalisé une enquête sérologique rétrospective provinciale anonymisée au moyen d'un échantillon randomisé stratifié de sérum résiduel soumis entre mai 2015 et août 2016 au dépistage systématique. Ils ont stratifié le processus de sélection selon l'âge, le sexe et la zone de santé régionale afin de garantir un échantillonnage proportionné. Pour déterminer la séroprévalence du VJC et du VLA, ils ont confirmé la positivité des résultats d'échantillons positifs au test ELISA au moyen de tests de séroneutralisation par réduction des plaques (TSRP). Résultats : Au total, 452 échantillons de sérum ont fait l'objet d'un dépistage. Au NB, la séroprévalence des anticorps anti-VSGC était évaluée à 31,6 % (IC à 95 %, 27,4 % à 36,1 %), pour 143 échantillons positifs. Selon les résultats du TSRP, la plupart des expositions à un seul virus étaient causées par le VJC (38 cas sur 143, 26,6 %) plutôt qu'au VLA (trois cas sur 143, 2,1 %). Les espèces de VSGC, auxquelles les 102 autres personnes séropositives ont été exposées, n'ont pas pu être établies avec précision. Conclusions : La prévalence d'exposition humaine au VSGC est élevée, mais comparable aux taux observés dans d'autres régions des provinces de l'Atlantique. Des études comme celle-ci fournissent des données épidémiologiques de référence importantes à l'égard du risque d'exposition humaine à ces arbovirus négligés. Il faut tenir compte du VLA et du VJC dans le diagnostic différentiel de maladie fébrile et neuro-invasive pendant la saison des moustiques, notamment lorsque les tests pour dépister d'autres étiologies courantes sont négatifs ou non concluants.
RESUMEN
BACKGROUND: Jamestown Canyon virus (JCV) is a mosquito-borne orthobunyavirus that causes acute febrile illness, meningitis, and meningoencephalitis, mainly among adults. JCV is widely distributed in North America and the number of JCV cases in the U.S. has increased in recent years. Therefore, the central nervous system disease caused by JCV can be considered a potentially re-emerging viral disease. However, the seroprevalence of JCV is unknown in Japan. The purpose of this study is to evaluate the seroprevalence of JCV in the Japanese population. METHODS: We used an IgG enzyme-linked immunosorbent assay (IgG-ELISA) with JCV-infected cell-lysates and/or a neutralizing (NT) antibody assay. The cut-off value of IgG-ELISA was determined using IgG-ELISA to analyze serum specimens from 37 healthy Japanese donors. IgG-ELISA was validated by assessing its sensitivity and specificity, using 38 human serum samples previously tested for the presence or absence of antibodies against JCV and snowshoe hare virus (SSHV), in an in-house NT antibody assay conducted by the Public Health Agency of Canada. The seroepidemiological study was performed using IgG-ELISA and NT antibody assay to analyze 246 human serum samples from the serum bank of the National Institute of Infectious Diseases (NIID) in Japan. RESULTS: The cut-off value of IgG-ELISA was determined at 0.20, based on the mean (- 0.075) and standard deviation (0.092) values using Japanese donors' sera. The sensitivity and the specificity of IgG-ELISA determined using 25 JCV-positive and 4 JCV-negative serum samples were 96 and 100%, respectively. Analysis of the 246 Japanese serum samples revealed that no specimen showed a higher value than the cut-off value of IgG-ELISA, and no sample tested positive by the NT antibody assay. CONCLUSIONS: Our results showed that JCV is not circulating significantly in Japan. To the best of our knowledge, this is the first report to demonstrate the seroprevalence of JCV in the general population in Japan.
Asunto(s)
Anticuerpos Antivirales/inmunología , Virus de la Encefalitis de California/inmunología , Encefalitis de California/epidemiología , Ensayo de Inmunoadsorción Enzimática/métodos , Pruebas de Neutralización/métodos , Adolescente , Adulto , Animales , Anticuerpos Neutralizantes/sangre , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/sangre , Niño , Preescolar , Culicidae/virología , Encefalitis de California/virología , Femenino , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Lactante , Recién Nacido , Japón/epidemiología , Masculino , Persona de Mediana Edad , Prevalencia , Sensibilidad y Especificidad , Estudios Seroepidemiológicos , Adulto JovenRESUMEN
Neutralizing antibodies against Zika virus (ZIKV) are a robust indicator of exposure to ZIKV. Here we describe the plaque reduction neutralization test (PRNT), which quantifies ZIKV-neutralizing antibodies (nAbs) by incubating infectious ZIKV with different dilutions of sera and applying the mixture to a susceptible Vero cell monolayer. After 4 days of incubation, neutral red dye is added to optimize the visualization of plaques caused by ZIKV cytopathic effect. Compared with controls, sera containing nAbs against ZIKV will demonstrate ≥90% reduction in the number of plaques observed. This protocol can be utilized for research applications and serological diagnostic testing with the use of additional controls.
Asunto(s)
Anticuerpos Antivirales/aislamiento & purificación , Pruebas de Neutralización/métodos , Ensayo de Placa Viral/métodos , Infección por el Virus Zika/diagnóstico , Virus Zika/inmunología , Animales , Anticuerpos Neutralizantes/metabolismo , Anticuerpos Antivirales/sangre , Especificidad de Anticuerpos , Chlorocebus aethiops , Grupos Control , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática/métodos , Humanos , Pruebas Serológicas/métodos , Células Vero , Infección por el Virus Zika/sangre , Infección por el Virus Zika/inmunología , Infección por el Virus Zika/virologíaRESUMEN
BACKGROUND: West Nile Virus (WNV) is a member of the Japanese Encephalitis (JE) serocomplex within the Flaviviridae family. We report four whole blood donors and one plasma donor with WNV nucleic acid test (NAT)-reactive donations between September 2018 and November 2019, following recent Japanese Encephalitis virus (JEV) vaccination. CASE SERIES: Cases 1 and 4 had reactive WNV NAT donations 1 day after receiving the JEV vaccine. Case 2 had a reactive WNV donation 3 days after receiving the JEV vaccine. Case 3 had a reactive WNV NAT donation 3 days after returning from Arizona and 1 day after receiving the JEV vaccine. Case 5 had a reactive WNV donation the same day as receiving the JEV vaccine. STUDY DESIGN AND METHODS: WNV screening used the Roche cobas WNV nucleic acid test (NAT) (Roche Molecular Systems). Reference testing on WNV-reactive donations was carried out by the National Microbiology Laboratory (NML). JEV vaccine dilutions were also analyzed. RESULTS: Supplemental NAT was negative for WNV and JEV for Cases 1, 3, and 5. Case 2 had a weak amplification curve for one of two JEV NAT targets. Case 4 was JEV NAT-positive, WNV NAT-negative. Serologic testing on donation specimens for Cases 2, 4, and 5 did not support recent or remote WNV infection. JEV vaccine dilutions were detected by both cobas and supplemental NAT. CONCLUSIONS: We recommend implementing a temporary blood donor deferral following a JEV vaccination, if screening utilizes a WNV assay with the capability of detecting other members of the JE serocomplex.
Asunto(s)
Donantes de Sangre , Virus de la Encefalitis Japonesa (Especie)/inmunología , Vacunación , Fiebre del Nilo Occidental/diagnóstico , Virus del Nilo Occidental/aislamiento & purificación , Adulto , Anciano , Reacciones Cruzadas , Femenino , Humanos , Masculino , Tamizaje Masivo/métodos , Persona de Mediana Edad , Técnicas de Amplificación de Ácido Nucleico , ARN Viral/análisis , ARN Viral/aislamiento & purificación , Vacunación/efectos adversos , Inactivación de Virus , Fiebre del Nilo Occidental/sangre , Fiebre del Nilo Occidental/etiología , Virus del Nilo Occidental/genética , Adulto JovenRESUMEN
Chikungunya virus (CHIKV) is a mosquito-borne pathogen that causes CHIKV fever. Definitive diagnosis is crucial for patients experiencing symptoms similar to other arboviral diseases because they can vary in clinical consequences. An increasing number of patients experience long-term rheumatic effects of CHIKV infection, but these cases may not be optimally detected by molecular assays and anti-CHIKV IgM ELISAs (M-ELISAs) used for confirmation and screening, respectively. The subsequent confirmatory serological test, the plaque reduction neutralization test (PRNT), is laborious and time-consuming. In this study, we evaluated a new diagnostic algorithm in which the M-ELISA is conducted in parallel with an anti-CHIKV IgG ELISA (G-ELISA) and observed that the Euroimmun M-ELISA combined with the Euroimmun G-ELISA or the Abcam G-ELISA exhibited excellent sensitivity and specificity for CHIKV. The combinations demonstrated perfect and near perfect inter-rater agreement with the PRNT, respectively, suggesting their potential to be used as alternatives to the confirmatory serological PRNT assay for CHIKV.
Asunto(s)
Anticuerpos Antiidiotipos/sangre , Anticuerpos Antivirales/sangre , Fiebre Chikungunya/diagnóstico , Ensayo de Inmunoadsorción Enzimática , Algoritmos , Fiebre Chikungunya/sangre , Fiebre Chikungunya/inmunología , Virus Chikungunya , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Juego de Reactivos para Diagnóstico/normas , Sensibilidad y Especificidad , Pruebas SerológicasRESUMEN
The previous serological algorithm for Zika virus (ZIKV) comprised screening by anti-ZIKV IgM capture ELISA (MAC-ELISA) for samples collected within 3 months postexposure or onset (MPEO). Samples positive by MAC-ELISA and samples collected beyond 3 MPEO were tested by the confirmatory plaque reduction neutralization test (PRNT), which proved laborious and time-consuming during the 2015 outbreak. Thus, we evaluated several ZIKV ELISAs to establish an anti-IgM and anti-IgG combination for use as a screening tool for all samples prior to PRNT confirmation. The MAC-ELISA or InBios-M in combination with the Euroimmun-G demonstrated sensitivities of 99.1% and 97.2%, respectively, and nonflavivirus specificity of 96.0%. Their cross-reactivities were 71.4% and 50.0%, respectively, for sera positive for Dengue virus antibodies. Due to near-perfect interrater agreement with PRNT and excellent detection of samples collected beyond 3 MPEO, these combinations were recommended as a screening protocol in a new high-throughput algorithm with special considerations for ZIKV diagnostics.
Asunto(s)
Algoritmos , Anticuerpos Antivirales/sangre , Tamizaje Masivo/métodos , Pruebas Serológicas/métodos , Infección por el Virus Zika/diagnóstico , Virus Zika/inmunología , Reacciones Cruzadas , Virus del Dengue/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Sensibilidad y EspecificidadRESUMEN
Due to the increase of Zika virus (ZIKV) transmission throughout the world, many commercial kits have recently become available to aid in laboratory diagnosis of ZIKV infections in clinical samples. Here, we analyze the fully automated Liaison® XL Zika Capture immunoglobulin M (IgM) assay against the recommended IgM-capture enzyme-linked immunosorbent assay.
Asunto(s)
Anticuerpos Antivirales/sangre , Inmunoglobulina M/sangre , Pruebas Serológicas/métodos , Infección por el Virus Zika/diagnóstico , Virus Zika/inmunología , Automatización de Laboratorios/métodos , HumanosRESUMEN
Using residual serum samples from Nova Scotia, Canada, we found that 87.8% of tested deer and an estimated 20.6% of the human population were infected with Jamestown Canyon virus. Human seropositivity reached 48.2% in 1 region. This virus may be an underrecognized cause of disease in Nova Scotia.
Asunto(s)
Ciervos , Virus de la Encefalitis de California/aislamiento & purificación , Encefalitis de California/veterinaria , Estudios Seroepidemiológicos , Adolescente , Adulto , Animales , Niño , Encefalitis de California/epidemiología , Encefalitis de California/virología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Nueva Escocia/epidemiología , Adulto JovenRESUMEN
California serogroup (CSG) viruses, such as Jamestown Canyon and snowshoe hare viruses, are mosquitoborne pathogens that cause febrile illness and neurologic disease. Human exposures have been described across Canada, but infections are likely underdiagnosed. We describe a case of neuroinvasive illness in a New Brunswick, Canada, patient infected with a CSG virus.
Asunto(s)
Disfunción Cognitiva/virología , Virus de la Encefalitis de California/clasificación , Encefalitis de California/epidemiología , Anticuerpos Antivirales/inmunología , Canadá/epidemiología , Disfunción Cognitiva/diagnóstico , Virus de la Encefalitis de California/inmunología , Encefalitis de California/diagnóstico , Encefalitis de California/transmisión , Encefalitis de California/virología , Historia del Siglo XXI , Humanos , Inmunoglobulina M/inmunología , Estudios Seroepidemiológicos , SerogrupoRESUMEN
Because of the global spread of Zika virus, accurate and high-throughput diagnostic immunoassays are needed. We compared the sensitivity and specificity of 5 commercially available Zika virus serologic assays to the recommended protocol of Zika virus IgM-capture ELISA and plaque-reduction neutralization tests. Most commercial immunoassays showed low sensitivity, which can be increased.
Asunto(s)
Inmunoensayo/métodos , Infección por el Virus Zika/virología , Virus Zika/aislamiento & purificación , Anticuerpos Antivirales/análisis , Anticuerpos Antivirales/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Humanos , Inmunoglobulina G/análisis , Inmunoglobulina G/inmunología , Inmunoglobulina M/análisis , Inmunoglobulina M/inmunología , Pruebas de Neutralización/métodos , Sensibilidad y Especificidad , Virus Zika/inmunología , Infección por el Virus Zika/diagnósticoRESUMEN
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.
Asunto(s)
Anticuerpos Antivirales/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Pruebas Serológicas/métodos , Infección por el Virus Zika/diagnóstico , Virus Zika/inmunología , Adulto , Estudios de Cohortes , Femenino , Humanos , Masculino , Persona de Mediana Edad , Ontario , Embarazo , Sensibilidad y EspecificidadRESUMEN
With the emerging Zika virus (ZIKV) epidemic, accessible real-time reverse transcription-PCR (rRT-PCR) assays are needed to streamline testing. The commercial Altona Diagnostics RealStar ZIKV rRT-PCR test kit (Altona PCR) has been approved for emergency use authorization by the U.S. FDA. Our aim was to verify the Altona PCR by comparing it to the CDC-designed dual-target ZIKV rRT-PCR reference assay (reference PCR) and describe the demographics of patients tested for ZIKV by rRT-PCR in Ontario, Canada. A large set of clinical specimens was tested for ZIKV by the Altona PCR and the reference PCR. Positive or equivocal specimens underwent PCR and Sanger sequencing targeting the ZIKV NS5 gene. A total of 671 serum specimens were tested by the reference PCR: 58 (8.6%) were positive, 193 (28.8%) were equivocal, and 420 (62.6%) were negative. Ninety percent of the reference PCR-positive patients were tested in the first 5 days after symptom onset. The Altona PCR was performed on 284/671 specimens tested by the reference PCR. The Altona PCR was positive for 53/58 (91%) reference PCR-positive specimens and 16/193 (8%) reference PCR-equivocal specimens; the ZIKV NS5 PCR was positive for all 68 Altona PCR-positive specimens and negative for all 181 Altona PCR-negative specimens that underwent the NS5 PCR. The Altona PCR has very good sensitivity (91%) and specificity (97%) compared to the reference PCR. The Altona PCR can be used for ZIKV diagnostic testing and has less extensive verification requirements than a laboratory-developed test.
Asunto(s)
Sangre/virología , Líquido Cefalorraquídeo/virología , ARN Viral/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Orina/virología , Infección por el Virus Zika/diagnóstico , Virus Zika/genética , Adulto , Reacciones Cruzadas , Pruebas Diagnósticas de Rutina/métodos , Femenino , Humanos , Masculino , Juego de Reactivos para Diagnóstico , Sensibilidad y Especificidad , Infección por el Virus Zika/virologíaRESUMEN
Arthropod-borne diseases negatively affect humans worldwide. Understanding the biology of the arthropod vectors and the pathogens they harbor, the arthropods are moving targets as a result of climate change, ecosystem degradation, species introductions, and increased human travel. Viruses within the California serogroup of the genus Orthobunyavirus (family Bunyaviridae) are among the mosquito-borne viruses of concern owing to their zoonotic potential. Two of these, snowshoe hare virus (SSHV) and Jamestown Canyon virus, were shown, using a combination of serology and virus isolations, to circulate on the Island of Newfoundland, Canada, in the 1980s. More recently, serological analysis demonstrated that these two viruses continue to circulate on the Island in several domesticated and wild animals. Here, we detected the seroconversion to SSHV in wild snowshoe hares and in a single sentinel rabbit. The seroconversion in the sentinel rabbit occurred in early August (2011), which corresponded to the weeks of peak mosquito collections and the timing of the detection of SSHV in suspected mosquito vectors. A portion of the SSHV S segment sequence was generated from mosquito pools collected at sites near the sentinel rabbits and phylogenetically analyzed using the neighbor-joining method with other available California serogroup virus sequences. This analysis validated the SSHV identification but showed that the Newfoundland sequence fell outside the other SSHV sequences available, which originated from the United States between 1959 and 2005.
Asunto(s)
Culicidae/virología , Virus de la Encefalitis de California/fisiología , Encefalitis de California/transmisión , Liebres , Insectos Vectores/virología , Animales , Encefalitis de California/virología , Terranova y Labrador , Estaciones del AñoRESUMEN
Studies examining the prevalence of zoonotic agents in the Caribbean are very limited. The objective of this study was to examine the seroprevalence of seven zoonotic agents among individuals residing on 10 English-speaking Caribbean countries. Sera from healthy, pregnant women were collected from Antigua-Barbuda, Belize, Bermuda, Dominica, Grenada, Jamaica, Montserrat, St. Kitts-Nevis, St. Lucia, and St. Vincent-Grenadines and tested for the presence of IgG antibodies to dengue virus, hepatitis E virus, hantaviruses, leptospiral agents, spotted fever group rickettsiae (SFGR), typhus group rickettsiae (TGR), and Coxiella burnetii (Q fever). The highest seroprevalence values were observed for dengue virus, SFGR, and leptospirosis, although the lowest seroprevalence values were observed for hepatitis E virus, C. burnetii, and TGR. Antibodies to hantaviruses were not detected in any individuals.