RESUMO
Beta-casein is a primary milk protein that constitutes approximately 30% of the casein in bovine milk, with the two most common types in cattle being A1 and A2. The A2 protein differs from the A1 version due to a mutation in the codon at position 67, resulting in a histidine to proline substitution. However, the bioactive peptide, beta-casomorphine-7 (BCM7), which originates from partial proteolysis of the A1 variant, has been linked to several gastrointestinal disorders in humans. Production of A1 beta casein-free products is increasing demand in the milk market, worldwide. This study generated and characterized a polyclonal IgY antibody that specifically recognizes the A1 beta-casein protein present in cow's milk. A commercially available IgY anti-A1 antibody was used as a positive control, and the sensitivity and specificity of both the commercial and produced anti-A1 antibodies were evaluated. The results showed 100% sensitivity and specificity of 100% of the commercial IgY anti-A1. The in-house produced anti-A1 antibody demonstrated a sensitivity of 95.2% and a specificity of 100%, indicating its potential as a reliable and cost effective tool for detecting A1 beta-casein protein in milk samples.
Assuntos
Caseínas , Leite , Humanos , Animais , Feminino , Bovinos , Leite/química , Anticorpos/análise , MutaçãoRESUMO
In resource-limited conditions such as the COVID-19 pandemic, on-site detection of diseases using the Point-of-care testing (POCT) technique is becoming a key factor in overcoming crises and saving lives. For practical POCT in the field, affordable, sensitive, and rapid medical testing should be performed on simple and portable platforms, instead of laboratory facilities. In this review, we introduce recent approaches to the detection of respiratory virus targets, analysis trends, and prospects. Respiratory viruses occur everywhere and are one of the most common and widely spreading infectious diseases in the human global society. Seasonal influenza, avian influenza, coronavirus, and COVID-19 are examples of such diseases. On-site detection and POCT for respiratory viruses are state-of-the-art technologies in this field and are commercially valuable global healthcare topics. Cutting-edge POCT techniques have focused on the detection of respiratory viruses for early diagnosis, prevention, and monitoring to protect against the spread of COVID-19. In particular, we highlight the application of sensing techniques to each platform to reveal the challenges of the development stage. Recent POCT approaches have been summarized in terms of principle, sensitivity, analysis time, and convenience for field applications. Based on the analysis of current states, we also suggest the remaining challenges and prospects for the use of the POCT technique for respiratory virus detection to improve our protection ability and prevent the next pandemic.
Assuntos
COVID-19 , Vírus , Humanos , Testes Imediatos , PandemiasRESUMO
The mosquito-borne disease caused by the Rocio virus is a neglected threat, and new immune inputs for serological testing are urgently required for diagnosis in low-resource settings and epidemiological surveillance. We used in silico approaches to identify a specific antigenic peptide (p_ROCV2) in the NS1 protein of the Rocio virus that was theoretically predicted to be stable and exposed on its surface, where it demonstrated key properties allowing it to interact with antibodies. These findings related to the molecular dynamics of this peptide provide important insights for advancing diagnostic platforms and investigating therapeutic alternatives.
Assuntos
Flavivirus , Simulação de Dinâmica Molecular , Animais , Testes Imunológicos , Simulação de Acoplamento Molecular , Peptídeos , Proteínas não Estruturais Virais/químicaRESUMO
This study demonstrates the influence of pregnancy on serum diagnosis of enzootic bovine leukosis (EBL), emphasizing the importance of routine testing to maintain herd health. For this, 143 pregnant cows were sampled in duplicate (30 days before and 15 days after calving). For EBL diagnosis, samples were submitted to agar gel immunodiffusion testing (AGID). Different results were observed before and after delivery in seventy-six serum samples (53.15%), indicating variations in the levels of serum globulins in the blood during the peripartum period. Therefore, using a single sample for serological diagnosis during the birth season might not represent the correct infection status of animal health due to physiological variations in antibody concentrations.
Assuntos
Leucose Enzoótica Bovina , Vírus da Leucemia Bovina , Animais , Anticorpos Antivirais , Bovinos , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Imunodifusão/métodos , Imunodifusão/veterinária , Período Periparto , GravidezRESUMO
Leprosy was recognized as a zoonotic disease, associated with nine-banded armadillos (Dasypus novemcinctus) in the Southern United States of America in 2011. In addition, there is growing evidence to support a role for armadillos in zoonotic leprosy in South America. The current study evaluated twenty specimens of the six-banded armadillo (Euphractus sexcinctus), collected from rural locations in the state of Rio Grande do Norte (RN), Brazil for evidence of infection with Mycobacterium leprae. Serum was examined using two "in-house" enzyme-linked immunosorbent assays (ELISAs) and via two commercially available (ML flow and NDO-LID®) immunochromatographic lateral flow (LF) tests, for detection of the PGL-I and/or LID-1 antigens of the bacterium. The presence of M. leprae DNA in liver tissue was examined using the multi-copy, M. leprae-specific repetitive element (RLEP), as target in conventional and nested PCR assays. Molecular and anti-PGL-I-ELISA data indicated that 20/20 (100 %) of the armadillos were infected with M. leprae. The corresponding detection levels recorded with the LF tests were 17/20 (85 %) and 16/20 (85 %), for the NDO-LID® and ML flow tests, respectively. Our results indicate that, in common with D. novemcinctus, six banded armadillos (a species hunted and reared as a food-source in some regions of Brazil, including RN), represent a potential reservoir of M. leprae and as such, their role in a possible zoonotic cycle of leprosy within Brazil warrants further investigation.
Assuntos
Tatus/microbiologia , Reservatórios de Doenças/veterinária , Hanseníase/veterinária , Mycobacterium leprae/genética , Mycobacterium leprae/imunologia , Animais , Brasil/epidemiologia , Reservatórios de Doenças/microbiologia , Ensaio de Imunoadsorção Enzimática , Feminino , Hanseníase/epidemiologia , Masculino , Reação em Cadeia da Polimerase , Zoonoses/epidemiologia , Zoonoses/microbiologiaRESUMO
BACKGROUND: A simple blood test for detecting active tuberculosis (TB) could be key to this epidemic containment, given that a large proportion of patients are unable to produce sputum for testing. Currently available interferon-γ release assays (IGRAs) are inadequate to diagnose active TB, with reported pooled sensitivity and specificity both under 81%. OBJECTIVE: To explore whether cytokines/chemokines other than interferon-γ in response to long-term cell stimulation could improve the ability to distinguish between different TB infection status. METHODS: We prospectively enrolled subjects with newly diagnosed pulmonary TB and their household contacts in Santiago. All contacts were tested with IGRA. Peripheral blood mononuclear cells were obtained and antigen-specific stimulated for 72â¯h before collecting their culture supernatants. RESULTS: Subjects with active TB displayed markedly low cytokines/chemokines secretion upon PBMC stimulation, with lower GM-CSF being the best differentiator from IGRA(+) contacts, with 71% (95% CI 53-85) sensitivity, 86% (95% CI 65-97) specificity and AUC = 0.79 (p = 0.0003). On the other hand, when compared to the uninfected IGRA(-) contacts, higher level of IL-2 secretion was the best indicator of active TB, with 73.5% (95% CI 56-87) sensitivity, 85% (95% CI 66-96) specificity and AUCâ¯=â¯0.79 (pâ¯=â¯0.0001). No single cytokine/chemokine released upon stimulation could accurately differentiate between active TB and all TB contacts grouped together. CONCLUSION: GM-CSF and IL-2 provided the best yield to differentiate active TB from latent TB and from TB uninfected, respectively, with higher specificities than that reported for IGRAs. However, none of both resulted sensitive enough to be used as a stand-alone biomarker for active TB.
Assuntos
Antígenos de Bactérias/imunologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Testes Imunológicos , Interleucina-2/metabolismo , Tuberculose Latente/diagnóstico , Leucócitos Mononucleares/metabolismo , Mycobacterium tuberculosis/imunologia , Tuberculose Pulmonar/diagnóstico , Adolescente , Adulto , Idoso , Biomarcadores/metabolismo , Chile , Busca de Comunicante , Diagnóstico Diferencial , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Interações Hospedeiro-Patógeno , Humanos , Interferon gama/imunologia , Interferon gama/metabolismo , Testes de Liberação de Interferon-gama , Interleucina-2/imunologia , Tuberculose Latente/imunologia , Tuberculose Latente/metabolismo , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/microbiologia , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Estudos Prospectivos , Reprodutibilidade dos Testes , Fatores de Tempo , Tuberculose Pulmonar/imunologia , Tuberculose Pulmonar/metabolismo , Adulto JovemRESUMO
The canine lymphoma rarely involves the nasal cavity. The diagnosis can be established by means of cytology, histology and immunohistochemistry, where the latter is a valuable technique for identification and classification of tumor cells. The study describes a case of intranasal lymphoma in a dog, with the aid of immunodiagnostic. A canine, male, seven years old mongrel, had volume growth in all nasal dorsal and lateral, in association with anterior uveitis. As a complementary exam was asked to cytology of lymph nodes and nasal tumor. The cytological evaluation of the lymph nodes showed the presence of amastigotes of Leishmania spp., while the cytology of nasal tumors exhibited morphological pattern indicative of lymphoma. The animal euthanasia was opted and same animal was submitted for necropsy. The autopsy findings included the presence of tumor to the fullest extent of the nasal cavities, causing complete luminal obliteration. Nasal tumor, liver and eye fragments were collected, which were sent for histopathological analysis, and which revealed neoplastic proliferation, consisting of round cells, and suggested conducting immunohistochemical analysis to confirm the diagnosis. The immunohistochemical profile applied detected a lymphoma cells Natural Killer. The intranasal lymphoma in dogs is often overlooked in the differential diagnosis of upper respiratory tract diseases. It is fundamental to the realization of immunohistochemical test, because it amounts to a valuable diagnostic tool for patients with lymphoma
O linfoma canino raramente envolve a cavidade nasal. O diagnóstico pode ser estabelecido por meio da citologia, histopatologia e imunoistoquímica, onde a ultima é uma técnica valiosa para a identificação e classificação de células tumorais. O trabalho objetivou descrever um caso de linfoma intranasal em um canino, com o auxílio do imunodiagnóstico. Um canino, macho, sete anos de idade, sem raça definida, possuía aumento de volume em toda região nasal dorsal e lateral, em associação com uveíte anterior. Como exame complementar solicitou-se citologia de linfonodos e da tumoração nasal. A avaliação citológica dos gânglios linfáticos evidenciou a presença de formas amastigotas de Leishmania spp., enquanto a citologia da tumoração nasal exibiu padrão morfológico indicativo de linfoma. Optou-se pela eutanásia do animal e o mesmo foi encaminhado para necropsia. Os achados necroscópicos incluíram a presença de tumor em toda a extensão das cavidades nasais, ocasionando completa obliteração luminal e perda total dos ossos endoturbinados e conchas nasais. Foram coletados fragmentos do tumor nasal, fígado e globo ocular, os quais foram encaminhados para análise histopatológica, a qual revelou proliferação neoplásica, composta por células redondas, sendo sugerida a realização de análise imunoistoquímica para confirmação diagnóstica. O perfil imunoistoquímico aplicado detectou um linfoma de células Natural Killer. O linfoma intranasal em cães muitas vezes é negligenciado no diagnóstico diferencial das enfermidades do trato respiratório superior. Torna-se fundamental a realização do ensaio imunoistoquímico, pois equivale a uma valiosa ferramenta diagnóstica para os pacientes com linfoma
Assuntos
Animais , Cães , Cavidade Nasal/patologia , Linfoma/patologia , Linfoma/veterinária , Imuno-Histoquímica/veterinária , Neoplasias Bucais/veterinária , Testes Imunológicos/veterinária , Técnicas Citológicas/veterináriaRESUMO
The canine lymphoma rarely involves the nasal cavity. The diagnosis can be established by means of cytology, histology and immunohistochemistry, where the latter is a valuable technique for identification and classification of tumor cells. The study describes a case of intranasal lymphoma in a dog, with the aid of immunodiagnostic. A canine, male, seven years old mongrel, had volume growth in all nasal dorsal and lateral, in association with anterior uveitis. As a complementary exam was asked to cytology of lymph nodes and nasal tumor. The cytological evaluation of the lymph nodes showed the presence of amastigotes of Leishmania spp., while the cytology of nasal tumors exhibited morphological pattern indicative of lymphoma. The animal euthanasia was opted and same animal was submitted for necropsy. The autopsy findings included the presence of tumor to the fullest extent of the nasal cavities, causing complete luminal obliteration. Nasal tumor, liver and eye fragments were collected, which were sent for histopathological analysis, and which revealed neoplastic proliferation, consisting of round cells, and suggested conducting immunohistochemical analysis to confirm the diagnosis. The immunohistochemical profile applied detected a lymphoma cells Natural Killer. The intranasal lymphoma in dogs is often overlooked in the differential diagnosis of upper respiratory tract diseases. It is fundamental to the realization of immunohistochemical test, because it amounts to a valuable diagnostic tool for patients with lymphoma(AU)
O linfoma canino raramente envolve a cavidade nasal. O diagnóstico pode ser estabelecido por meio da citologia, histopatologia e imunoistoquímica, onde a ultima é uma técnica valiosa para a identificação e classificação de células tumorais. O trabalho objetivou descrever um caso de linfoma intranasal em um canino, com o auxílio do imunodiagnóstico. Um canino, macho, sete anos de idade, sem raça definida, possuía aumento de volume em toda região nasal dorsal e lateral, em associação com uveíte anterior. Como exame complementar solicitou-se citologia de linfonodos e da tumoração nasal. A avaliação citológica dos gânglios linfáticos evidenciou a presença de formas amastigotas de Leishmania spp., enquanto a citologia da tumoração nasal exibiu padrão morfológico indicativo de linfoma. Optou-se pela eutanásia do animal e o mesmo foi encaminhado para necropsia. Os achados necroscópicos incluíram a presença de tumor em toda a extensão das cavidades nasais, ocasionando completa obliteração luminal e perda total dos ossos endoturbinados e conchas nasais. Foram coletados fragmentos do tumor nasal, fígado e globo ocular, os quais foram encaminhados para análise histopatológica, a qual revelou proliferação neoplásica, composta por células redondas, sendo sugerida a realização de análise imunoistoquímica para confirmação diagnóstica. O perfil imunoistoquímico aplicado detectou um linfoma de células Natural Killer. O linfoma intranasal em cães muitas vezes é negligenciado no diagnóstico diferencial das enfermidades do trato respiratório superior. Torna-se fundamental a realização do ensaio imunoistoquímico, pois equivale a uma valiosa ferramenta diagnóstica para os pacientes com linfoma(AU)
Assuntos
Animais , Cães , Linfoma/patologia , Linfoma/veterinária , Cavidade Nasal/patologia , Testes Imunológicos/veterinária , Neoplasias Bucais/veterinária , Técnicas Citológicas/veterinária , Imuno-Histoquímica/veterináriaRESUMO
Background: Sporotrichosis is an endemic zoonosis in the metropolitan region of Rio de Janeiro caused by fungi included in the Sporothrix complex, in which cats are the main source of infection for humans and animals. Coinfections in cats with sporotrichosis from this region, their risk factors and how they affect the treatment outcome in these animals are little known. The objectives of this study were to determine the coinfections of Sporothrix spp. with Toxoplasma gondii, Feline Immunodeficiency Virus (FIV) and Feline Leukemia Virus (FeLV) and to correlate these infections with risk factors and the outcome of sporotrichosis treatment in cats from an endemic area of sporotrichosis in Rio de Janeiro, Brazil.Materials, Methods & Results: It was conducted a cohort study involving 213 cats with definitive diagnosis of sporotrichosis from the metropolitan area of Rio de Janeiro and assisted in the Laboratory of Clinical Research on Dermatozoonosis in Domestic Animals (LAPCLIN-DERMZOO)/Evandro Chagas National Institute of Infectious Diseases (INI)/Oswaldo Cruz Foundation (Fiocruz), in Rio de Janeiro, RJ, Brazil, from November 2007 until February 2011. These animals were monthly evaluated due to sporotrichosis treatment until their sporotrichosis treatment outcomes. In every clinical evaluation, 5 mL of blood were collected in order to obtain the serum, which was stored at -20ºC. Information from the animals medical records have also been collected, such as sex, eating habits, living with other cats, access to the streets, castration, age and the outcome of sporotrichosis treatment. Serological follow-up of anti-T. gondii antibodies were performed through indirect hemagglutination assay (IHA) and indirect fluorescent antibody test (IFAT) in all clinical evaluations. The FIV and FeLV antibody detection were made through a rapid immunoassay using the catsserum samples from the first clinical evaluation.[...]
Assuntos
Animais , Gatos , Coinfecção/veterinária , Esporotricose/veterinária , Fatores de Risco , Toxoplasma , Vírus da Imunodeficiência Felina , Vírus da Leucemia Felina , Brasil , Doenças Endêmicas/veterinária , Retroviridae , Sporothrix , Testes Imunológicos/veterináriaRESUMO
Background: Sporotrichosis is an endemic zoonosis in the metropolitan region of Rio de Janeiro caused by fungi included in the Sporothrix complex, in which cats are the main source of infection for humans and animals. Coinfections in cats with sporotrichosis from this region, their risk factors and how they affect the treatment outcome in these animals are little known. The objectives of this study were to determine the coinfections of Sporothrix spp. with Toxoplasma gondii, Feline Immunodeficiency Virus (FIV) and Feline Leukemia Virus (FeLV) and to correlate these infections with risk factors and the outcome of sporotrichosis treatment in cats from an endemic area of sporotrichosis in Rio de Janeiro, Brazil.Materials, Methods & Results: It was conducted a cohort study involving 213 cats with definitive diagnosis of sporotrichosis from the metropolitan area of Rio de Janeiro and assisted in the Laboratory of Clinical Research on Dermatozoonosis in Domestic Animals (LAPCLIN-DERMZOO)/Evandro Chagas National Institute of Infectious Diseases (INI)/Oswaldo Cruz Foundation (Fiocruz), in Rio de Janeiro, RJ, Brazil, from November 2007 until February 2011. These animals were monthly evaluated due to sporotrichosis treatment until their sporotrichosis treatment outcomes. In every clinical evaluation, 5 mL of blood were collected in order to obtain the serum, which was stored at -20ºC. Information from the animals medical records have also been collected, such as sex, eating habits, living with other cats, access to the streets, castration, age and the outcome of sporotrichosis treatment. Serological follow-up of anti-T. gondii antibodies were performed through indirect hemagglutination assay (IHA) and indirect fluorescent antibody test (IFAT) in all clinical evaluations. The FIV and FeLV antibody detection were made through a rapid immunoassay using the catsserum samples from the first clinical evaluation.[...](AU)
Assuntos
Animais , Gatos , Esporotricose/veterinária , Coinfecção/veterinária , Toxoplasma , Vírus da Imunodeficiência Felina , Vírus da Leucemia Felina , Fatores de Risco , Sporothrix , Testes Imunológicos/veterinária , Retroviridae , Brasil , Doenças Endêmicas/veterináriaRESUMO
Ethnic origin, genetics, gender and environmental factors have been shown to influence some immunologic indices, so that development of reference values for populations of different backgrounds may be necessary. We have determined the distribution of lymphocyte subsets in healthy Brazilian individuals from birth to adulthood. Lymphocyte subsets were determined using four-colour cytometry in a cross-sectional study of 463 human immunodeficiency virus-unexposed children and adults from birth through 49 years of age. Lymphocyte subsets varied according to age, as previously observed in other studies. However, total CD4+ T cell numbers were lower than what was described in the Pediatric AIDS Clinical Trials Group P1009 (PACTG P1009), which assessed an American population of predominantly African and Hispanic backgrounds until the 12-18 year age range, when values were comparable. Naïve percentages and absolute values of CD8+ T cells, as assessed by CD45RA expression, were also lower than the PACTG P1009 data for all analysed age ranges. CD38 expression on both CD4+ and CD8+ T cells was lower than the PACTG P1009 values, with a widening gap between the two studies at older age ranges. Different patterns of cell differentiation seem to occur in different settings and may have characteristic expression within each population.
Assuntos
Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Adulto Jovem , /citologia , /citologia , Subpopulações de Linfócitos/citologia , Fatores Etários , Linfócitos B/citologia , Brasil , Ensaios Clínicos como Assunto , Estudos Transversais , Citometria de Fluxo/métodos , Voluntários Saudáveis , Células Matadoras Naturais/citologia , Contagem de Linfócitos , Leucócitos Mononucleares/citologia , Valores de ReferênciaRESUMO
Toxocariasis is a neglected zoonosis caused by the nematodes Toxocara canis and Toxocara cati. This disease is widespread in many countries, reaching high prevalence independently of the economic conditions. However, the true number of cases of toxocariasis is likely to be underestimated owing to the lack of adequate surveillance programs. Although some diagnostic tests are available, their sensitivity and specificity need to be improved. In addition, treatment options for toxocariasis are limited and are non-specific. Toxocariasis is listed as one of the five most important neglected diseases by the CDC. This review presents recent advances related to the control of toxocariasis, including new immunodiagnostics, therapies, and drug formulations, as well as novel interventions using DNA vaccines, immunomodulators, and probiotics.
Assuntos
Toxocaríase/diagnóstico , Toxocaríase/terapia , Animais , Humanos , Fatores Imunológicos/uso terapêutico , Testes Imunológicos/tendências , Doenças Negligenciadas/diagnóstico , Doenças Negligenciadas/patologia , Doenças Negligenciadas/prevenção & controle , Doenças Negligenciadas/terapia , Probióticos , Toxocaríase/patologia , Toxocaríase/prevenção & controle , Toxocaríase/transmissão , Vacinas de DNARESUMO
Immunoglobulin Y (IgY) is the major antibody isotype in birds, reptiles, amphibia, and lungfish, playing a similar biological role as mammal IgG. Due to its phylogenetic distance, immune diversification and presence in the egg yolk, IgY provide a number of advantages in immunodiagnostic compared to IgG from mammals. Moreover, IgY production is in agreement with international efforts to reduce, refine and if possible, to replace animals in experimentation, contributing substantially in favor of animal welfare. This article presents an overview about structural and functional features, production and applications of IgY in immunodiagnostic, as well as the advantages of chicken antibodies use.
A imunoglobulina Y (IgY) é a classe de anticorpos de maior importância em aves, répteis, anfíbios e peixes pulmonados, desempenhando um papel semelhante a IgG de mamíferos. Devido a sua distância filogenética, mecanismos de diversificação imune e presença na gema do ovo, IgY proporciona uma série de vantagens em imunodiagnóstico, quando comparada a IgG de mamíferos. Além disso, esse método alternativo de produção de anticorpo está de acordo com os esforços internacionais para reduzir, refinar e, se possível, substituir animais em experimentação, contribuindo substancialmente a favor do bem-estar animal. Este artigo apresenta uma revisão sobre as características estruturais e funcionais da IgY, bem como os métodos de produção, vantagens e aplicações em imunodiagnóstico, além das vantagens da sua utilização.
RESUMO
Immunoglobulin Y (IgY) is the major antibody isotype in birds, reptiles, amphibia, and lungfish, playing a similar biological role as mammal IgG. Due to its phylogenetic distance, immune diversification and presence in the egg yolk, IgY provide a number of advantages in immunodiagnostic compared to IgG from mammals. Moreover, IgY production is in agreement with international efforts to reduce, refine and if possible, to replace animals in experimentation, contributing substantially in favor of animal welfare. This article presents an overview about structural and functional features, production and applications of IgY in immunodiagnostic, as well as the advantages of chicken antibodies use.(AU)
A imunoglobulina Y (IgY) é a classe de anticorpos de maior importância em aves, répteis, anfíbios e peixes pulmonados, desempenhando um papel semelhante a IgG de mamíferos. Devido a sua distância filogenética, mecanismos de diversificação imune e presença na gema do ovo, IgY proporciona uma série de vantagens em imunodiagnóstico, quando comparada a IgG de mamíferos. Além disso, esse método alternativo de produção de anticorpo está de acordo com os esforços internacionais para reduzir, refinar e, se possível, substituir animais em experimentação, contribuindo substancialmente a favor do bem-estar animal. Este artigo apresenta uma revisão sobre as características estruturais e funcionais da IgY, bem como os métodos de produção, vantagens e aplicações em imunodiagnóstico, além das vantagens da sua utilização.(AU)