RESUMO
Andes orthohantavirus (ANDV) is the etiologic agent of hantavirus cardiopulmonary syndrome (HCPS), which has a case fatality rate around 35%, with no effective treatment or vaccine available. ANDV neutralizing antibody (NAb) measurements are important for the evaluation of the immune response following infection, vaccination, or passive administration of investigational monoclonal or polyclonal antibodies. The standard assay for NAb measurement is a focus reduction neutralization test (FRNT) featuring live ANDV and must be completed under biosafety level (BSL)-3 conditions. In this study, we compared neutralization assays featuring infectious ANDV or vesicular stomatitis virus (VSV) pseudovirions decorated with ANDV glycoproteins for their ability to measure anti-ANDV NAbs from patient samples. Our studies demonstrate that VSV pseudovirions effectively measure NAb from clinical samples and have greater sensitivity compared to FRNT with live ANDV. Importantly, the pseudovirus assay requires less labor and sample materials and can be conducted at BSL-2.
Assuntos
Infecções por Hantavirus , Orthohantavírus , Anticorpos Neutralizantes , Anticorpos Antivirais , Infecções por Hantavirus/diagnóstico , Humanos , Testes de NeutralizaçãoRESUMO
This work describes the first method using biochar (BC) as carbonaceous platform for immunoassay application. BC is a highly functionalized material obtained through biomass pyrolysis under controlled conditions. Due to the highly functionalized surface, covalent binding between BC and biomolecules can be performed by EDC/NHS conjugation. The application of the modified electrode was done with Hantavirus, that are etiologic agents mainly transmitted by wild rodents. Among its pathologies Hantavirus Cardiopulmonary Syndrome (HCPS) arises at Americas, caused by Hantavirus Araucária and reaches 40% lethality. The diagnostic is based on the presence of specific hantavirus nucleoprotein (Np), under viremic condition or IgG2b antibodies (Ab), during first symptoms. The results presented a device sensitivity of 5.28⯵A dec-1 and a LOD of 0.14â¯ngâ¯mL-1 to the Np detection, ranging from 5.0â¯ngâ¯mL-1 to 1.0⯵gâ¯mL-1, the Ab detection works as qualitative type sensor above 200â¯ngâ¯mL-1. Both sensors were evaluated its selectivity and serum samples; selectivity against Gumboro disease, VP2 protein, and antibody IgG2a against Yellow fever disease (YF), respectively. So, the devices here proposed are promising tool suitable for both rodent and human hantavirus clinical surveys.
Assuntos
Carvão Vegetal/química , Orthohantavírus/isolamento & purificação , Anticorpos Imobilizados/imunologia , Anticorpos Antivirais/imunologia , Sangue/virologia , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Eletrodos , Orthohantavírus/química , Humanos , Imunoensaio/métodos , Imunoglobulina G/imunologia , Limite de Detecção , Reprodutibilidade dos Testes , Proteínas Virais/imunologiaRESUMO
Hantaviruses are etiologic agents of a zoonotic disease transmitted mainly from wild rodents to humans, causing Hemorrhagic Fever with Renal Syndrome in Eurasia and the Hantavirus Cardiopulmonary Syndrome in the Americas (HCPS), reaching a lethality rate of 40% in Brazil. Hantavirus diagnostic and seroprevalence are often based on the presence of IgM and IgG antibodies against the virus. Here we propose a rapid test assay able to identify hantavirus antibodies with sensibility and specificity similar to ELISA assays. We analyzed five groups of samples, including healthy human population and small mammals of endemic areas, suspected cases of HCPS, patients with non-related infections and a serum panel from a different geographical region. The test presented good rates of sensibility (87-100%) and specificity (97-100%) for all groups, being a promising tool suitable for both rodent and human hantavirus epidemiological surveys.
Assuntos
Anticorpos Antivirais/sangue , Infecções por Hantavirus/epidemiologia , Infecções por Hantavirus/veterinária , Imunoensaio/métodos , Orthohantavírus/imunologia , Sistemas Automatizados de Assistência Junto ao Leito , Animais , Humanos , Roedores , Sensibilidade e Especificidade , Estudos SoroepidemiológicosRESUMO
En Chile, la infección por hantavirus Andes (ANDV) tiene una expresión clínica variable, reconociéndose diversos grados de severidad. El presente estudio se realizó con el objeto de analizar la posible asociación entre la constitución genética de pacientes chilenos para el sistema HLA y la expresión clínica de la infección por ANDV. Se analizaron los alelos HLA A, B, DRB1 y DQB1, en dos grupos de pacientes con infección por ANDV: 41 pacientes con evolución clínica leve (sin insuficiencia respiratoria severa y sin requerimientos de ventilación mecánica) y 46 pacientes con evolución clínica grave (con insuficiencia respiratoria grave y/o shock). La determinación molecular del sistema HLA se realizó mediante SSP-PCR. El alelo HLA DRB1 * 15, se encontró en una frecuencia significativamente más alta en los pacientes leves (p = 0,007). Por lo tanto, el alelo DRB 1*15 se asociaría al curso clínico leve de la enfermedad. El alelo HLA-B*08, se encontró en una frecuencia mayor en los pacientes graves, la diferencia alcanzó una significación estadística marginal (p = 0,06). Así, el alelo HLA-B*08, podría estar asociado al curso clínico grave de síndrome cardiopulmonar ocasionado por hantavirus Andes.
Andes hantavirus (ANDV) infection in Chile has a variable clinical expression, and infected individuals may present with different grades of disease severity. This study aimed to determine if clinical expression of ANDV infection in Chilean patients is associated with the HLA system. HLA alíeles A, B, DRB1 and DQB1, were studied in two groups of patients with confirmed ANDV infection: 41 patients with a mild disease course (without respiratory failure and cardiovascular shock) and 46 patients with a severe disease course (with respiratory failure and shock). Molecular typing of HLA system was performed by SSP-PCR. The HLA-DRB 1*15 alíele, was significantly more common in the group of patients with mild disease (p = 0,007) and thus for possibly associated with a protective effect against ANDV infection. Conversely, HLA-B*08 was more common in patients with severe disease (p = 0,06). Although the association was marginally significant, alíele HLA-B*08 may be linked to an increased susceptibility to the severe clinical course of HCPS by ANDV.