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1.
Rev. bras. oftalmol ; 83: e0003, 2024. tab, graf
Artículo en Portugués | LILACS | ID: biblio-1529931

RESUMEN

RESUMO Objetivo: Identificar o perfil dos doadores de tecidos oculares humanos na área de atuação do Banco de Olhos da Paraíba, destacando o impacto da sorologia positiva para hepatite B no descarte dos tecidos para transplante. Métodos: O estudo é transversal e utilizou dados do Banco de Olhos da Paraíba entre janeiro de 2013 e dezembro de 2022. Dados sobre procedência, idade, sexo, causa do óbito, tempo entre óbito e enucleação, resultados sorológicos e motivo de descarte das córneas dos doadores foram coletados. Resultados: O maior motivo de descarte foi por sorologia positiva (56,5%), sendo positivadas as sorologias positivas para hepatite B e HBsAg em 11,1% e 4,75% dos pacientes, respectivamente. Conclusão: A sorologia positiva para hepatite B como um critério de descarte absoluto é responsável por grande parcela de descartes, apesar da pouca informação sobre suas repercussões e representação de infectividade nos receptores do transplante.


ABSTRACT Objective: To identify the profile of human ocular tissue donors in the area covered by the Eye Bank of Paraíba (PB), highlighting the impact of positive serology for hepatitis B (anti-HBc) in the disposal of tissues for transplantation. Methods: This is a cross-sectional that uses data from the Eye Bank of Paraíba (PB) between January 2013 and December 2022. Data on origin, age, sex, cause of death, time between death and enucleation, serological results, and reason for discarded donor corneas were collected. Results: The main reason for discarding was due to positive serology (56.5%), with positive anti-HBc and HBsAg serology in 11.1% and 4.75% of patients, respectively. Conclusion: Anti-HBc positive serology as an absolute disposal criterion is responsible for great part of disposals, despite little information about its repercussions and representation of infectivity in transplant recipients.


Asunto(s)
Humanos , Masculino , Femenino , Preescolar , Niño , Adolescente , Adulto , Persona de Mediana Edad , Anciano , Anciano de 80 o más Años , Donantes de Tejidos/estadística & datos numéricos , Trasplante de Córnea/normas , Trasplante de Córnea/estadística & datos numéricos , Selección de Donante/normas , Bancos de Ojos/normas , Anticuerpos contra la Hepatitis B/análisis , Pruebas Serológicas/normas , Virus de la Hepatitis B , Estudios Transversales , Estudios Retrospectivos , Transmisión de Enfermedad Infecciosa/legislación & jurisprudencia , Transmisión de Enfermedad Infecciosa/prevención & control , Transmisión de Enfermedad Infecciosa/estadística & datos numéricos , Bancos de Ojos/estadística & datos numéricos , Hepatitis B/prevención & control , Hepatitis B/transmisión , Antígenos del Núcleo de la Hepatitis B/análisis
2.
J Clin Microbiol ; 61(6): e0011223, 2023 06 20.
Artículo en Inglés | MEDLINE | ID: mdl-37222630

RESUMEN

Treponema pallidum subsp. pallidum is a fastidious spirochete and the etiologic agent of syphilis, a sexually transmitted infection (STI). Syphilis diagnoses and disease staging are based on clinical findings and serologic testing. Moreover, according to most international guidelines, PCR analysis of swab samples from genital ulcers is included in the screening algorithm where possible. It has been suggested that PCR might be omitted from the screening algorithm due to low added value. As an alternative to PCR, IgM serology might be used. In this study, we wanted to establish the added value of PCR and IgM serology for diagnosing primary syphilis. Added value was defined as finding more cases of syphilis, preventing overtreatment, or limiting the extent of partner notification to more recent partners. We found that both PCR and IgM immunoblotting could aid the timely diagnosis of early syphilis in ~24% to 27% of patients. PCR has the greatest sensitivity and can be applied to cases with an ulcer with suspected reinfection or primary infection. In the absence of lesions, the IgM immunoblot could be used. However, the IgM immunoblot has better performance in cases with suspected primary infection than in reinfections. The target population, testing algorithm, time pressures, and costs should determine whether either test provides sufficient value to be implemented in clinical practice.


Asunto(s)
Pruebas Diagnósticas de Rutina , Inmunoglobulina M , Sífilis , Humanos , Immunoblotting/normas , Inmunoglobulina M/análisis , Reacción en Cadena de la Polimerasa/normas , Sífilis/diagnóstico , Sífilis/inmunología , Sífilis/microbiología , Treponema pallidum/genética , Pruebas Serológicas/normas , Pruebas Diagnósticas de Rutina/economía , Pruebas Diagnósticas de Rutina/métodos , Pruebas Diagnósticas de Rutina/normas , Sensibilidad y Especificidad
3.
Int J Mol Sci ; 23(6)2022 Mar 17.
Artículo en Inglés | MEDLINE | ID: mdl-35328649

RESUMEN

As the global SARS-CoV-2 pandemic continues to plague healthcare systems, it has become clear that opportunistic pathogens cause a considerable proportion of SARS-CoV-2-associated mortality and morbidity cases. Of these, Covid-Associated Pulmonary Aspergilliosis (CAPA) is a major concern with evidence that it occurs in the absence of traditional risk factors such as neutropenia and is diagnostically challenging for the attending physician. In this review, we focus on the immunopathology of SARS-CoV-2 and how this potentiates CAPA through dysregulation of local and systemic immunity as well as the unintended consequences of approved COVID treatments including corticosteroids and IL-6 inhibitors. Finally, we will consider how knowledge of the above may aid in the diagnosis of CAPA using current diagnostics and what treatment should be instituted in probable and confirmed cases.


Asunto(s)
COVID-19/complicaciones , COVID-19/inmunología , Susceptibilidad a Enfermedades/inmunología , Interacciones Huésped-Patógeno/inmunología , Aspergilosis Pulmonar/etiología , SARS-CoV-2/inmunología , Antifúngicos/uso terapéutico , Biomarcadores , COVID-19/virología , Manejo de la Enfermedad , Humanos , Huésped Inmunocomprometido , Aspergilosis Pulmonar/diagnóstico , Aspergilosis Pulmonar/terapia , Reproducibilidad de los Resultados , Pruebas Serológicas/métodos , Pruebas Serológicas/normas , Resultado del Tratamiento
4.
PLoS Negl Trop Dis ; 16(2): e0010152, 2022 02.
Artículo en Inglés | MEDLINE | ID: mdl-35120141

RESUMEN

BACKGROUND: Chikungunya virus (CHIKV) causes febrile illnesses and has always been misdiagnosed as other viral infections, such as dengue and Zika; thus, a laboratory test is needed. Serological tests are commonly used to diagnose CHIKV infection, but their accuracy is questionable due to varying degrees of reported sensitivities and specificities. Herein, we conducted a systematic review and meta-analysis to evaluate the diagnostic accuracy of serological tests currently available for CHIKV. METHODOLOGY AND PRINCIPAL FINDINGS: A literature search was performed in PubMed, CINAHL Complete, and Scopus databases from the 1st December 2020 until 22nd April 2021. Studies reporting sensitivity and specificity of serological tests against CHIKV that used whole blood, serum, or plasma were included. QUADAS-2 tool was used to assess the risk of bias and applicability, while R software was used for statistical analyses. Thirty-five studies were included in this meta-analysis; 72 index test data were extracted and analysed. Rapid and ELISA-based antigen tests had a pooled sensitivity of 85.8% and 82.2%, respectively, and a pooled specificity of 96.1% and 96.0%, respectively. According to our meta-analysis, antigen detection tests serve as a good diagnostic test for acute-phase samples. The IgM detection tests had more than 90% diagnostic accuracy for ELISA-based tests, immunofluorescence assays, in-house developed tests, and samples collected after seven days of symptom onset. Conversely, low sensitivity was found for the IgM rapid test (42.3%), commercial test (78.6%), and for samples collected less than seven of symptom onset (26.2%). Although IgM antibodies start to develop on day 2 of CHIKV infection, our meta-analysis revealed that the IgM detection test is not recommended for acute-phase samples. The diagnostic performance of the IgG detection tests was more than 93% regardless of the test formats and whether the test was commercially available or developed in-house. The use of samples collected after seven days of symptom onset for the IgG detection test suggests that IgG antibodies can be detected in the convalescent-phase samples. Additionally, we evaluated commercial IgM and IgG tests for CHIKV and found that ELISA-based and IFA commercial tests manufactured by Euroimmun (Lübeck, Germany), Abcam (Cambridge, UK), and Inbios (Seattle, WA) had diagnostic accuracy of above 90%, which was similar to the manufacturers' claim. CONCLUSION: Based on our meta-analysis, antigen or antibody-based serological tests can be used to diagnose CHIKV reliably, depending on the time of sample collection. The antigen detection tests serve as a good diagnostic test for samples collected during the acute phase (≤7 days post symptom onset) of CHIKV infection. Likewise, IgM and IgG detection tests can be used for samples collected in the convalescent phase (>7 days post symptom onset). In correlation to the clinical presentation of the patients, the combination of the IgM and IgG tests can differentiate recent and past infections.


Asunto(s)
Antígenos Virales/aislamiento & purificación , Fiebre Chikungunya/diagnóstico , Pruebas Serológicas/normas , Antígenos Virales/sangre , Virus Chikungunya/inmunología , Virus Chikungunya/aislamiento & purificación , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Sensibilidad y Especificidad
5.
Clin Transl Med ; 11(12): e668, 2021 12.
Artículo en Inglés | MEDLINE | ID: mdl-34898055

RESUMEN

The level of postvaccine protection depends on two factors: antibodies and T-cell responses. While the first one is relatively easily measured, the measuring of the second one is a difficult problem. The recent studies indicate that the first one may be a good proxy for the protection, at least for SARS-CoV-2. The massive data currently gathered by both researcher and citizen scientists may be pivotal in confirming this observation, and the collective body of evidence is growing daily. This leads to an acceptance of IgG antibody levels as an accessible biomarker of individual's protection. With enormous and immediate need for assessing patient condition at the point of care, quantitative antibody analysis remains the most effective and efficient way to assess the protection against the disease. Let us not discount importance of reference points in the turmoil of current pandemics.


Asunto(s)
Anticuerpos Antivirales/química , Anticuerpos/química , Biomarcadores/metabolismo , COVID-19/sangre , COVID-19/inmunología , Especificidad de Anticuerpos , Humanos , Sistema Inmunológico , Inmunidad , Inmunoglobulina G/metabolismo , Unidades de Cuidados Intensivos , Pandemias , Sistemas de Atención de Punto , SARS-CoV-2 , Pruebas Serológicas/métodos , Pruebas Serológicas/normas , Vacunas
6.
Viruses ; 13(9)2021 08 30.
Artículo en Inglés | MEDLINE | ID: mdl-34578299

RESUMEN

Phleboviruses (genus Phlebovirus, family Phenuiviridae) are emerging pathogens of humans and animals. Sand-fly-transmitted phleboviruses are found in Europe, Africa, the Middle East, and the Americas, and are responsible for febrile illness and nervous system infections in humans. Rio Grande virus (RGV) is the only reported phlebovirus in the United States. Isolated in Texas from southern plains woodrats, RGV is not known to be pathogenic to humans or domestic animals, but serologic evidence suggests that sheep (Ovis aries) and horses (Equus caballus) in this region have been infected. Rift Valley fever virus (RVFV), a phlebovirus of Africa, is an important pathogen of wild and domestic ruminants, and can also infect humans with the potential to cause severe disease. The introduction of RVFV into North America could greatly impact U.S. livestock and human health, and the development of vaccines and countermeasures is a focus of both the CDC and USDA. We investigated the potential for serologic reagents used in RVFV diagnostic assays to also detect cells infected with RGV. Western blots and immunocytochemistry assays were used to compare the antibody detection of RGV, RVFV, and two other New World phlebovirus, Punta Toro virus (South and Central America) and Anhanga virus (Brazil). Antigenic cross-reactions were found using published RVFV diagnostic reagents. These findings will help to inform test interpretation to avoid false positive RVFV diagnoses that could lead to public health concerns and economically costly agriculture regulatory responses, including quarantine and trade restrictions.


Asunto(s)
Reacciones Cruzadas/inmunología , Phlebovirus/inmunología , Juego de Reactivos para Diagnóstico/normas , Virus de la Fiebre del Valle del Rift/inmunología , Pruebas Serológicas/normas , Animales , Infecciones por Bunyaviridae/clasificación , Infecciones por Bunyaviridae/diagnóstico , Infecciones por Bunyaviridae/inmunología , Caballos/virología , Phlebovirus/clasificación , Phlebovirus/patogenicidad , Fiebre del Valle del Rift/diagnóstico , Fiebre del Valle del Rift/inmunología , Virus de la Fiebre del Valle del Rift/patogenicidad , Pruebas Serológicas/métodos , Ovinos/virología , Estados Unidos
7.
Int J Technol Assess Health Care ; 37(1): e87, 2021 Sep 22.
Artículo en Inglés | MEDLINE | ID: mdl-34548114

RESUMEN

OBJECTIVE: In vitro diagnostic tests for SARS-COV-2, also known as serological tests, have rapidly spread. However, to date, mostly single-center technical and diagnostic performance's assessments have been carried out without an intralaboratory validation process and a health technology assessment (HTA) systematic approach. Therefore, the rapid HTA for evaluating antibody tests for SARS-COV-2 was applied. METHODS: The use of rapid HTA is an opportunity to test innovative technology. Unlike traditional HTA (which evaluates the benefits of new technologies after being tested in clinical trials or have been applied in practice for some time), the rapid HTA is performed during the early stages of developing new technology. A multidisciplinary team conducted the rapid HTA following the HTA Core Model® (version 3.0) developed by the European Network for Health Technology Assessment. RESULTS: The three methodological and analytical steps used in the HTA applied to the evaluation of antibody tests for SARS-COV-2 are reported: the selection of the tests to be evaluated; the research and collection of information to support the adoption and appropriateness of the technology; and the preparation of the final reports and their dissemination. Finally, the rapid HTA of serological tests for SARS-CoV-2 is summarized in a report that allows its dissemination and communication. CONCLUSIONS: The rapid-HTA evaluation method, in addition to highlighting the characteristics that differentiate the tests from each other, guarantees a timely and appropriate evaluation, becoming a tool to create a direct link between science and health management.


Asunto(s)
Prueba de COVID-19/métodos , COVID-19/diagnóstico , COVID-19/inmunología , Pruebas Serológicas/métodos , Humanos , SARS-CoV-2 , Pruebas Serológicas/normas , Evaluación de la Tecnología Biomédica , Factores de Tiempo
8.
Diagn Microbiol Infect Dis ; 101(3): 115412, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34425450

RESUMEN

The COVID-19 pandemic has led to the influx of immunoassays for the detection of antibodies towards severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) into the global market. The Canadian Public Health Laboratory Network Serology Task Force undertook a nationwide evaluation of twelve laboratory and 6 point-of-care based commercial serological assays for the detection of SARS-CoV-2 antibodies. We determined that there was considerable variability in the performance of individual tests and that an orthogonal testing algorithm should be prioritized to maximize the accuracy and comparability of results across the country. The manual enzyme immunoassays and point-of-care tests evaluated had lower specificity and increased coefficients of variation compared to automated enzyme immunoassays platforms putting into question their utility for large-scale sero-surveillance. Overall, the data presented here provide a comprehensive approach for applying accurate serological assays for longitudinal sero-surveillance and vaccine trials while informing Canadian public health policy.


Asunto(s)
Anticuerpos Antivirales/sangre , COVID-19/epidemiología , Laboratorios/normas , Salud Pública , SARS-CoV-2/inmunología , Pruebas Serológicas/normas , COVID-19/sangre , Canadá/epidemiología , Ensayos Analíticos de Alto Rendimiento , Humanos , Técnicas para Inmunoenzimas , SARS-CoV-2/aislamiento & purificación , Pruebas Serológicas/métodos
9.
J Med Virol ; 93(11): 6309-6316, 2021 11.
Artículo en Inglés | MEDLINE | ID: mdl-34375007

RESUMEN

Serological tests detecting antibodies for Epstein-Barr virus (EBV) antigens are frequently used to define infection status. Several new automated assays are available for this purpose. We compared the performance of Architect, Immulite, Vidas, and Euroimmune immunofluorescence assays (IFA)/enzyme-linked immunosorbent assays (ELISA) for the detection of EBV viral capsid antigen (VCA) immunoglobulin M (IgM), VCA IgG, Epstein-Barr nuclear antigen (EBNA)-1 IgG. The routine diagnosis of EBV in our laboratory is done by anti-EBV VCA IgM IFT, anti-EBV VCA IgG IFT, and anti-EBNA-1 IgG ELISA (Euroimmune) Kits. Samples were tested with EBV Kits of Architect, Immulite, and Vidas for anti-VCA IgM, anti-VCA IgG, and anti-EBNA-1 IgG. The agreement between assays was calculated for each marker individually and for the determination of the EBV infection profile, based on the combination of three markers. BIOCHIP Sequence EBV (Avidity test) and/or EUROLINE EBV Profile 2 (IgG/IgM) were used as confirmatory assays to resolve discrepancies. The best concordance for VCA IgM detection was between Immulite and Vidas; for VCA IgG and EBNA-1 IgG were between Architect and Vidas. The sensitivities and specificities for VCA IgM were 97% and 88% for IFA, 100% and 94% for Architect, 100% and 99% for Vidas, and 100% and 100% for Immulite, respectively. The most problematic marker was EBNA-1 IgG with a 68.1% specificity by Immulite. Vidas panel had a perfect performance (100%) for determining all EBV profiles. Overall, evaluated assays had comparable performance. There were more discordant VCA IgG and EBNA-1 IgG results than VCA IgM results. The agreement between Architect and Vidas was better than other assays.


Asunto(s)
Anticuerpos Antivirales/sangre , Infecciones por Virus de Epstein-Barr/sangre , Infecciones por Virus de Epstein-Barr/diagnóstico , Herpesvirus Humano 4/inmunología , Juego de Reactivos para Diagnóstico/normas , Pruebas Serológicas/normas , Adolescente , Adulto , Antígenos Virales/inmunología , Proteínas de la Cápside/inmunología , Niño , Preescolar , Infecciones por Virus de Epstein-Barr/inmunología , Femenino , Humanos , Inmunoglobulina M/sangre , Lactante , Mediciones Luminiscentes/instrumentación , Mediciones Luminiscentes/normas , Masculino , Persona de Mediana Edad , Sensibilidad y Especificidad , Pruebas Serológicas/instrumentación , Pruebas Serológicas/métodos , Adulto Joven
10.
Viruses ; 13(8)2021 07 31.
Artículo en Inglés | MEDLINE | ID: mdl-34452381

RESUMEN

The case fatality rate of rabies, nearly 100%, is one of the most unique characteristic of this ancient virus infection. The crucial role rabies virus neutralizing antibody plays in protection is both well established and explanation of why rabies serology is important. Various laboratory methods can and have been used but serum neutralization methods have long been the gold standard due to the ability to measure function (neutralization), however these methods can be difficult to perform for several reasons. Assays such as enzyme linked absorbance assays (ELISA), indirect fluorescence antibody (IFA) and more recently lateral flow methods are in use. Interpretation of results can be problematic, not only between methods but also due to modifications of the same method that can lead to misinterpretations. A common assumption in review of laboratory test results is that different methods for the same component produce comparable results under all conditions or circumstances. Assumptions and misinterpretations provide the potential for detrimental decisions, ranging from regulatory to clinically related, and most importantly what 'level' is protective. Review of the common challenges in performance and interpretation of rabies serology and specific examples illuminate critical issues to consider when reviewing and applying results of rabies serological testing.


Asunto(s)
Anticuerpos Antivirales/sangre , Virus de la Rabia/inmunología , Rabia/diagnóstico , Rabia/inmunología , Pruebas Serológicas/normas , Anticuerpos Neutralizantes/sangre , Interpretación Estadística de Datos , Ensayo de Inmunoadsorción Enzimática/métodos , Ensayo de Inmunoadsorción Enzimática/normas , Técnica del Anticuerpo Fluorescente Indirecta , Pruebas Hematológicas , Humanos , Pruebas de Neutralización/métodos , Pruebas de Neutralización/normas , Virus de la Rabia/aislamiento & purificación , Pruebas Serológicas/clasificación , Pruebas Serológicas/métodos
11.
Diagn Microbiol Infect Dis ; 101(3): 115489, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34352435

RESUMEN

BACKGROUND: Parvovirus B19 is a pathogenic virus often diagnosed by serology, yet little is known about analytical performance of commercial enzyme immunoassays (EIAs). OBJECTIVE: To investigate performance of 4 EIAs for parvovirus B19 IgM and IgG: Liaison, Euroimmun, Mikrogen and Virion/Serion. STUDY DESIGN: To compare 4 EIAs to Biotrin's ELISA on 168 samples and determine consensus score for discordant samples using Mikrogen's confirmatory line assay. RESULTS: Two thirds of results for IgM/IgG were identical for all 4 EIAs and Biotrin. Liaison shows the highest IgM sensitivity, but has low specificity. Euroimmun lacks IgM sensitivity. Mikrogen had a good overall performance, but had the lowest IgG specificity. Virion/Serion had variable performance with a low IgM specificity and the most borderline and cross-reactive results. CONCLUSIONS: Liaison and Mikrogen have similar performance to Biotrin's ELISA. Euroimmun lacks sensitivity and Virion/Serion produced many borderline and cross-reactive results.


Asunto(s)
Anticuerpos Antivirales/sangre , Eritema Infeccioso/diagnóstico , Técnicas para Inmunoenzimas/normas , Parvovirus B19 Humano/inmunología , Pruebas Serológicas/normas , Eritema Infeccioso/inmunología , Humanos , Técnicas para Inmunoenzimas/métodos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Sensibilidad y Especificidad , Pruebas Serológicas/métodos
12.
Parasit Vectors ; 14(1): 408, 2021 Aug 17.
Artículo en Inglés | MEDLINE | ID: mdl-34404476

RESUMEN

BACKGROUND: Toxoplasmosis caused by Toxoplasma gondii is a serious disease threatening human and animal health. People can be infected with T. gondii by ingesting raw pork contaminated with cysts or oocysts. Serological test is a sensitive and specific method usually used for large-scale diagnosis of T. gondii infection in humans and animals (such as pigs). Commercial pig Toxoplasma antibody ELISA diagnostic kits are expensive, which limits their use; moreover, the wide antigen composition used in these diagnostic kits is still unclear and difficult to standardize. The multiepitope peptide antigen is a novel diagnostic marker, and it has potential to be developed into more accurate and inexpensive diagnostic kits. METHODS: The synthetic multiepitope antigen (MAG) cDNA encoding a protein with epitopes from five T. gondii-dominant antigens (SAG1, GRA1, ROP2, GRA4, and MIC3) was designed, synthesized, and expressed in Escherichia coli BL21 (DE3) strain. The recombinant protein was detected through western blot with pig anti-T. gondii-positive and -negative serum, and then IgG enzyme-linked immunosorbent assay (ELISA) named MAG-ELISA was designed. The MAG-ELISA was evaluated in terms of specificity, sensitivity, and stability. The MAG-ELISA was also compared with a commercial PrioCHECK® Toxoplasma Ab porcine ELISA (PrioCHECK ELISA). Finally, the trend of pig anti-T. gondii IgG levels after artificial infection with RH tachyzoites was evaluated using MAG-ELISA and two other ELISA methods (rMIC3-ELISA and PrioCHECK ELISA). RESULTS: MAG antigen could be specifically recognized by pig anti-T. gondii-positive but not -negative serum. MAG-ELISA showed high diagnostic performance in terms of specificity (88.6%) and sensitivity (79.1%). MAG-ELISA could be used for detecting anti-T. gondii IgG in the early stage of T. gondii infection in pigs (at least 7 days after artificial infection). CONCLUSIONS: Our results suggest that MAG antigen can be applied to specifically recognize anti-T. gondii IgG in pig, and MAG-ELISA has the potential for large-scale screening tests of T. gondii infection in pig farms and intensive industries.


Asunto(s)
Antígenos de Protozoos/genética , Antígenos de Protozoos/inmunología , Epítopos/genética , Proteínas Recombinantes/inmunología , Pruebas Serológicas/normas , Toxoplasma/inmunología , Toxoplasmosis Animal/diagnóstico , Animales , Epítopos/inmunología , Inmunoglobulina G/sangre , Proteínas Recombinantes/genética , Sensibilidad y Especificidad , Pruebas Serológicas/métodos , Porcinos , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/inmunología , Enfermedades de los Porcinos/parasitología , Toxoplasma/genética , Toxoplasmosis Animal/sangre , Toxoplasmosis Animal/inmunología
13.
Viruses ; 13(7)2021 06 22.
Artículo en Inglés | MEDLINE | ID: mdl-34206519

RESUMEN

Alphaviruses have a single-stranded, positive-sense RNA genome that contains two open reading frames encoding either the non-structural or the structural genes. Upon infection, the genomic RNA is translated into the non-structural proteins (nsPs). NsPs are required for viral RNA replication and transcription driven from the subgenomic promoter (sgP). Transfection of an RNA encoding the luciferase gene under the control of the sgP into cells enabled the detection of replication-competent chikungunya virus (CHIKV) or Mayaro virus (MAYV) with high sensitivity as a function of the induced luciferase activity. This assay principle was additionally used to analyze virus-neutralizing antibodies in sera and might be an alternative to standard virus neutralization assays based on virus titration or the use of genetically modified tagged viruses.


Asunto(s)
Alphavirus/inmunología , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/sangre , ARN Viral/genética , Pruebas Serológicas/métodos , Alphavirus/clasificación , Infecciones por Alphavirus/sangre , Infecciones por Alphavirus/diagnóstico , Infecciones por Alphavirus/inmunología , Animales , Línea Celular , Virus Chikungunya/genética , Reacciones Cruzadas , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Luciferasas/genética , Ratones , Ratones Endogámicos BALB C , Sensibilidad y Especificidad , Pruebas Serológicas/normas
14.
PLoS Negl Trop Dis ; 15(6): e0009457, 2021 06.
Artículo en Inglés | MEDLINE | ID: mdl-34181665

RESUMEN

BACKGROUND: Serological surveys with multiplex bead assays can be used to assess seroprevalence to multiple pathogens simultaneously. However, multiple methods have been used to generate cut-off values for seropositivity and these may lead to inconsistent interpretation of results. A literature review was conducted to describe the methods used to determine cut-off values for data generated by multiplex bead assays. METHODOLOGY/PRINCIPAL FINDINGS: A search was conducted in PubMed that included articles published from January 2010 to January 2020, and 308 relevant articles were identified that included the terms "serology", "cut-offs", and "multiplex bead assays". After application of exclusion of articles not relevant to neglected tropical diseases (NTD), vaccine preventable diseases (VPD), or malaria, 55 articles were examined based on their relevance to NTD or VPD. The most frequently applied approaches to determine seropositivity included the use of presumed unexposed populations, mixture models, receiver operating curves (ROC), and international standards. Other methods included the use of quantiles, pre-exposed endemic cohorts, and visual inflection points. CONCLUSIONS/SIGNIFICANCE: For disease control programmes, seropositivity is a practical and easily interpretable health metric but determining appropriate cut-offs for positivity can be challenging. Considerations for optimal cut-off approaches should include factors such as methods recommended by previous research, transmission dynamics, and the immunological backgrounds of the population. In the absence of international standards for estimating seropositivity in a population, the use of consistent methods that align with individual disease epidemiological data will improve comparability between settings and enable the assessment of changes over time.


Asunto(s)
Enfermedades Transmisibles/sangre , Enfermedades Transmisibles/diagnóstico , Estudios Seroepidemiológicos , Pruebas Serológicas/métodos , Pruebas Serológicas/normas , Medicina Tropical/métodos , Humanos , Enfermedades Prevenibles por Vacunación/diagnóstico
16.
J Immunol Methods ; 496: 113088, 2021 09.
Artículo en Inglés | MEDLINE | ID: mdl-34181967

RESUMEN

Blue eye disease (BED) of pigs was identified in the early 1980s in La Piedad, Michoacan, Mexico. The causal agent is Porcine orthorubulavirus (PRV), which affects pigs of all ages, producing nervous, respiratory, and reproductive disorders. BED is geographically endemic to the center of Mexico, where 75% of the country's swine industry is concentrated. Due to its adverse effects on the swine industry and the risk of dissemination to other countries, it is essential to have reliable diagnostic methods for BED. The objective of this study was to establish the optimal conditions for three serological tests, hemagglutination inhibition (HI), immunoperoxidase monolayer assay (IPMA), and serum neutralization (SN), and to compare their sensitivity, specificity, kappa coefficient, and predictive values. Twelve different HI protocols (9408 tests), one SN protocol and one IPMA protocol (784 tests, each) were evaluated. Forty-nine sera were analyzed, and thirty-seven sera showed true positive results, while twelve showed true negative results. The kappa coefficient was used to assess the variation in each test. The best HI protocol registered a sensitivity and specificity of 89 and 100%, respectively, the IPMA test showed values of 85 and 100%, and the SN test registered a sensitivity of 91% and a specificity of 96%. One of the disadvantages of the HI test is that when chicken red blood cells (RBCs) are used, elution occurs in a short incubation time, which would decrease the specificity. The use of bovine RBCs increases the specificity of the testy and makes it more stable, but it decreases the sensitivity. The results of HI and SN revealed the importance of eliminating the complement system of the serum and removing other inhibitors to avoid test nonspecificity. The IPMA test does not use an active virus; hence, it is considered safe and does not present any risk of disseminating PRV.


Asunto(s)
Anticuerpos Neutralizantes/sangre , Anticuerpos Antivirales/sangre , Infecciones Virales del Ojo/diagnóstico , Pruebas de Inhibición de Hemaglutinación/veterinaria , Técnicas para Inmunoenzimas/veterinaria , Infecciones por Rubulavirus/diagnóstico , Rubulavirus/inmunología , Pruebas Serológicas/veterinaria , Enfermedades de los Porcinos/diagnóstico , Animales , Biomarcadores/sangre , Infecciones Virales del Ojo/sangre , Infecciones Virales del Ojo/inmunología , Infecciones Virales del Ojo/virología , Pruebas de Inhibición de Hemaglutinación/normas , Técnicas para Inmunoenzimas/normas , México , Valor Predictivo de las Pruebas , Reproducibilidad de los Resultados , Infecciones por Rubulavirus/sangre , Infecciones por Rubulavirus/inmunología , Infecciones por Rubulavirus/virología , Pruebas Serológicas/normas , Porcinos , Enfermedades de los Porcinos/sangre , Enfermedades de los Porcinos/inmunología , Enfermedades de los Porcinos/virología
17.
Transfusion ; 61(9): 2677-2687, 2021 09.
Artículo en Inglés | MEDLINE | ID: mdl-34121205

RESUMEN

BACKGROUND: Antibody response duration following severe acute respiratory syndrome coronavirus 2 infection tends to be variable and depends on severity of disease and method of detection. STUDY DESIGN AND METHODS: COVID-19 convalescent plasma from 18 donors was collected longitudinally for a maximum of 63-129 days following resolution of symptoms. All the samples were initially screened by the Ortho total Ig test to confirm positivity and subsequently tested with seven additional direct sandwich or indirect binding assays (Ortho, Roche, Abbott, Broad Institute) directed against a variety of antigen targets (S1, receptor binding domain, and nucleocapsid [NC]), along with two neutralization assays (Broad Institute live virus PRNT and Vitalant Research Institute [VRI] Pseudovirus reporter viral particle neutralization [RVPN]). RESULTS: The direct detection assays (Ortho total Ig total and Roche total Ig) showed increasing levels of antibodies over the time period, in contrast to the indirect IgG assays that showed a decline. Neutralization assays also demonstrated declining responses; the VRI RVPN pseudovirus had a greater rate of decline than the Broad PRNT live virus assay. DISCUSSION: These data show that in addition to variable individual responses and associations with disease severity, the detection assay chosen contributes to the heterogeneous results in antibody stability over time. Depending on the scope of the research, one assay may be preferable over another. For serosurveillance studies, direct, double Ag-sandwich assays appear to be the best choice due to their stability; in particular, algorithms that include both S1- and NC-based assays can help reduce the rate of false-positivity and discriminate between natural infection and vaccine-derived seroreactivity.


Asunto(s)
Anticuerpos Antivirales/inmunología , Donantes de Sangre , COVID-19/epidemiología , COVID-19/inmunología , SARS-CoV-2/inmunología , Anticuerpos Neutralizantes/sangre , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/sangre , COVID-19/sangre , COVID-19/diagnóstico , Interacciones Huésped-Patógeno/inmunología , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Estudios Seroepidemiológicos , Pruebas Serológicas/métodos , Pruebas Serológicas/normas , Índice de Severidad de la Enfermedad
18.
PLoS One ; 16(5): e0251263, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34010292

RESUMEN

Rift Valley fever virus (RVFV), an arbovirus belonging to the Phlebovirus genus of the Phenuiviridae family, causes the zoonotic and mosquito-borne RVF. The virus, which primarily affects livestock (ruminants and camels) and humans, is at the origin of recent major outbreaks across the African continent (Mauritania, Libya, Sudan), and in the South-Western Indian Ocean (SWIO) islands (Mayotte). In order to be better prepared for upcoming outbreaks, to predict its introduction in RVFV unscathed countries, and to run efficient surveillance programmes, the priority is harmonising and improving the diagnostic capacity of endemic countries and/or countries considered to be at risk of RVF. A serological inter-laboratory proficiency test (PT) was implemented to assess the capacity of veterinary laboratories to detect antibodies against RVFV. A total of 18 laboratories in 13 countries in the Middle East, North Africa, South Africa, and the Indian Ocean participated in the initiative. Two commercial kits and two in-house serological assays for the detection of RVFV specific IgG antibodies were tested. Sixteen of the 18 participating laboratories (88.9%) used commercial kits, the analytical performance of test sensitivity and specificity based on the seroneutralisation test considered as the reference was 100%. The results obtained by the laboratories which used the in-house assay were correct in only one of the two criteria (either sensitivity or specificity). In conclusion, most of the laboratories performed well in detecting RVFV specific IgG antibodies and can therefore be considered to be prepared. Three laboratories in three countries need to improve their detection capacities. Our study demonstrates the importance of conducting regular proficiency tests to evaluate the level of preparedness of countries and of building a network of competent laboratories in terms of laboratory diagnosis to better face future emerging diseases in emergency conditions.


Asunto(s)
Fiebre del Valle del Rift/diagnóstico , África/epidemiología , Animales , Anticuerpos Antivirales/sangre , Enfermedades Endémicas/veterinaria , Ensayo de Inmunoadsorción Enzimática/normas , Ensayo de Inmunoadsorción Enzimática/estadística & datos numéricos , Ensayo de Inmunoadsorción Enzimática/veterinaria , Humanos , Inmunoglobulina G/sangre , Océano Índico/epidemiología , Laboratorios/normas , Medio Oriente/epidemiología , Garantía de la Calidad de Atención de Salud , Reproducibilidad de los Resultados , Fiebre del Valle del Rift/epidemiología , Fiebre del Valle del Rift/inmunología , Virus de la Fiebre del Valle del Rift/inmunología , Factores de Riesgo , Pruebas Serológicas/normas , Pruebas Serológicas/estadística & datos numéricos , Pruebas Serológicas/veterinaria
19.
Viruses ; 13(4)2021 03 31.
Artículo en Inglés | MEDLINE | ID: mdl-33807442

RESUMEN

Flaviviruses circulate worldwide and cause a number of medically relevant human diseases, such as dengue, Zika, yellow fever, and tick-borne encephalitis (TBE). Serology plays an important role in the diagnosis of flavivirus infections, but can be impeded by antigenic cross-reactivities among flaviviruses. Therefore, serological diagnosis of a recent infection can be insufficiently specific, especially in areas where flaviviruses co-circulate and/or vaccination coverage against certain flaviviruses is high. In this study, we developed a new IgM assay format, which is well suited for the specific diagnosis of TBE, Zika and dengue virus infections. In the case of TBE and Zika, the IgM response proved to be highly specific for the infecting virus. In contrast, primary dengue virus infections induced substantial amounts of cross-reactive IgM antibodies, which is most likely explained by structural peculiarities of dengue virus particles. Despite the presence of cross-reactive IgM, the standardized nature and the quantitative read-out of the assay even allowed the serotype-specific diagnosis of recent dengue virus infections in most instances.


Asunto(s)
Anticuerpos Antivirales/sangre , Antígenos Virales/inmunología , Reacciones Cruzadas/inmunología , Infecciones por Flavivirus/diagnóstico , Flavivirus/inmunología , Inmunoglobulina M/sangre , Pruebas Serológicas/métodos , Antígenos Virales/clasificación , Estudios de Cohortes , Dengue/sangre , Dengue/diagnóstico , Dengue/inmunología , Virus del Dengue/inmunología , Virus de la Encefalitis Transmitidos por Garrapatas/inmunología , Encefalitis Transmitida por Garrapatas/diagnóstico , Encefalitis Transmitida por Garrapatas/inmunología , Flavivirus/clasificación , Infecciones por Flavivirus/sangre , Infecciones por Flavivirus/virología , Humanos , Serogrupo , Pruebas Serológicas/normas , Virus Zika/inmunología , Infección por el Virus Zika/sangre , Infección por el Virus Zika/diagnóstico , Infección por el Virus Zika/inmunología
20.
Diagn Microbiol Infect Dis ; 100(3): 115371, 2021 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-33838594

RESUMEN

We validated a multiplex bead assay for diphtheria toxoid IgG antibodies against the Vero cell toxin neutralization test using 1300 specimens (correlation = 0.88). At the ≥0.01 IU/mL cutoff for minimal seroprotection, sensitivity was 95% and specificity was 83%. Agreement for three categories (<0.01, 0.01-<0.1, ≥0.1 IU/mL) was 81% (kappa = 0.71).


Asunto(s)
Anticuerpos Antibacterianos/sangre , Toxoide Diftérico , Inmunoglobulina G/sangre , Pruebas Serológicas/métodos , Animales , Chlorocebus aethiops , Pruebas de Neutralización/métodos , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Pruebas Serológicas/normas , Células Vero
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