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1.
Anticancer Res ; 41(4): 2183-2186, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33813431

RESUMO

BACKGROUND/AIM: The aim of this study was to identify simple and reliable factors to detect clinically insignificant prostate cancer (PC) for avoiding immediate prostate biopsies using biparametric magnetic resonance imaging (MRI), which consists of T2-weighted and diffusion-weighted imaging. PATIENTS AND METHODS: We retrospectively evaluated 427 men with suspected PC, who underwent biparametric MRI and standard 12-core transrectal prostate biopsy. MRI and prostate specific antigen density (PSAD) were analysed. To evaluate the combination of the two parameters, patients were divided into three groups (Group A: MRI negative and PSAD <0.23, Group B: MRI positive or PSAD ≥0.23, Group C: MRI positive and PSAD ≥0.23). A grade of ≥2 was defined as clinically significant PC. RESULTS: Clinically significant PC was detected in 46.5% of men with positive MRI findings, and 60.0% of men with PSAD ≥0.23. When combining MRI and PSAD, detection rates of clinically significant PC were 10.0%, 28.4% and 65.3% in group A, B and, C, respectively. CONCLUSION: Negative biparametric MRI findings with PSAD <0.23 might be a reliable evidence for avoiding immediate prostate biopsies.


Assuntos
Imagem por Ressonância Magnética/métodos , Antígeno Prostático Específico/análise , Neoplasias da Próstata/diagnóstico , Neoplasias da Próstata/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia com Agulha de Grande Calibre/métodos , Imagem de Difusão por Ressonância Magnética , Humanos , Biópsia Guiada por Imagem , Testes Imunológicos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Cuidados Pré-Operatórios , Próstata/patologia , Antígeno Prostático Específico/sangue , Neoplasias da Próstata/metabolismo , Estudos Retrospectivos , Carga Tumoral , Ultrassonografia de Intervenção
2.
Medicine (Baltimore) ; 100(8): e24615, 2021 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-33663071

RESUMO

ABSTRACT: The T-SPOT.TB assay detects cellular immune responses to 2 core Mycobacterium tuberculosis antigens, early secreted antigenic target of 6-kDa protein (ESAT-6) and culture filtrate protein-10 (CFP-10). T-SPOT.TB has been recently used for auxiliary diagnosis of active pulmonary tuberculosis (PTB). However, testing can produce inconsistent results due to differential PTB patient immune responses to these antigens, prompting us to identify factors underlying inconsistent results.Data were retrospectively analyzed from 1225 confirmed PTB patients who underwent T-SPOT.TB testing at 5 specialized tuberculosis hospitals in China between December 2012 and November 2015. Numbers of spot-forming cells (SFCs) reflecting T cell responses to ESAT-6 and CFP-10 antigens were recorded then analyzed via multivariable logistic regression to reveal factors underlying discordant T cell responses to these antigens.The agreement rate of 84.98% (82.85%-86.94%) between PTB patient ESAT-6 and CFP-10 responses demonstrated high concordance. Additionally, positivity rates were higher for ESAT-6 than for CFP-10 (84.8% vs 80.7%, P < .001), with ESAT-6 and CFP-10 microwell SFC numbers for each single positive group not differing significantly (P > .99), while spot numbers of the single positive group were lower than numbers for the double positive group (P < .001). Elderly patients (aged ≥66 years) and patients receiving retreatment were most likely to have discordance results.ESAT-6 promoted significantly more positive T-SPOT.TB results than did CFP-10 in PTB patients. Advanced age and retreatment status were correlated with discordant ESAT-6 and CFP-10 results. Assessment of factors underlying discordance may lead to improved PTB diagnosis using T-SPOT.TB.


Assuntos
Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Fragmentos de Peptídeos/imunologia , Linfócitos T/imunologia , Tuberculose Pulmonar/imunologia , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , China , Feminino , Humanos , Imunidade Celular/imunologia , Testes Imunológicos/normas , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Sensibilidade e Especificidade , Adulto Jovem
3.
Lancet Child Adolesc Health ; 5(4): 284-294, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33600774

RESUMO

Childhood cancer and its treatment often impact the haematopoietic and lymphatic systems, with immunological consequences. Immunological assessments are not routinely included in surveillance guidelines for most survivors of childhood cancer, although a robust body of literature describes immunological outcomes, testing recommendations, and revaccination guidelines after allogeneic haematopoietic cell transplantation. Survivorship care providers might not fully consider the impaired recovery of a child's immune system after cancer treatment if the child has not undergone haematopoietic cell transplantation. We did a scoping review to collate the existing literature describing immune function after childhood cancer therapy, including both standard-dose chemotherapy and high-dose chemotherapy with haematopoietic cell rescue. This Review aims to summarise: the principles of immunology and testing of immune function; the body of literature describing immunological outcomes after childhood cancer therapy, with an emphasis on the risk of infection, when is testing indicated, and preventive strategies; and knowledge gaps and opportunities for future research.


Assuntos
Antineoplásicos/efeitos adversos , Sobreviventes de Câncer , Reconstituição Imune/imunologia , Síndromes de Imunodeficiência/imunologia , Neoplasias/terapia , Doenças Preveníveis por Vacina/prevenção & controle , Vacinas/uso terapêutico , Imunidade Adaptativa/imunologia , Contagem de Células Sanguíneas , Transplante de Células-Tronco Hematopoéticas , Humanos , Imunidade Inata/imunologia , Síndromes de Imunodeficiência/etiologia , Testes Imunológicos , Baço/imunologia
4.
Mem Inst Oswaldo Cruz ; 115: e200287, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33533869

RESUMO

BACKGROUND: The heat-labile nature of Dengue virus (DENV) in serum samples must be considered when applying routine diagnostic tests to avoid issues that could impact the accuracy of test results with direct implications for case management and disease reporting. OBJECTIVES: To check if pre-analytical variables, such as storage time and temperature, have an impact on the accuracy of the main routine diagnostic tests for dengue. METHODS: Virus isolation, reverse transcription real-time polymerase chain reaction (RT-PCR) and NS1 enzyme-linked immunosorbent assay (ELISA) were evaluated using 84 samples submitted to different pre-analytical conditions. FINDINGS: Sensitivity and negative predictive value were directly affected by sample storage conditions. RT-PCR and virus isolation showed greater dependence on well-conserved samples for an accurate diagnosis. Interestingly, even storage at -30ºC for a relatively short time (15 days) was not adequate for accurate results using virus isolation and RT-PCR tests. On the other hand, NS1 ELISA showed no significant reduction in positivity for aliquots tested under the same conditions as in the previous tests. MAIN CONCLUSIONS: Our results support the stability of the NS1 marker in ELISA diagnosis and indicate that the accuracy of routine tests such as virus isolation and RT-PCR is significantly affected by inadequate transport and storage conditions of serum samples.


Assuntos
Antígenos Virais/sangue , Vírus da Dengue/isolamento & purificação , Dengue/diagnóstico , Ensaio de Imunoadsorção Enzimática/métodos , Testes Imunológicos/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Proteínas não Estruturais Virais/imunologia , Anticorpos Antivirais/sangue , Antígenos Virais/imunologia , Dengue/sangue , Dengue/virologia , Vírus da Dengue/genética , Vírus da Dengue/imunologia , Humanos , Valor Preditivo dos Testes , Sensibilidade e Especificidade , Proteínas não Estruturais Virais/genética
5.
Med Clin North Am ; 105(2): 387-396, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33589110

RESUMO

Systemic lupus erythematosus (SLE) is an autoimmune inflammatory condition that may involve multiple organ systems. Although the antinuclear antibody (ANA) test is positive in nearly every case of SLE, it is not specific for this disease and must be interpreted in the appropriate clinical context. Key features that warrant ANA testing include unexplained multisystem inflammatory disease, symmetric joint pain with inflammatory features, photosensitive rash, and cytopenias. ANA staining patterns and more specific autoantibody testing may be helpful in diagnosis of suspected SLE or ANA-associated disease. For patients with nonspecific symptoms, such as malaise and fatigue, ANA testing is of limited value.


Assuntos
Anticorpos Antinucleares/sangue , Lúpus Eritematoso Sistêmico , Humanos , Testes Imunológicos/métodos , Lúpus Eritematoso Sistêmico/diagnóstico , Lúpus Eritematoso Sistêmico/imunologia
6.
Allergol. immunopatol ; 49(1): 95-100, ene.-feb. 2021. tab
Artigo em Inglês | IBECS | ID: ibc-199231

RESUMO

INTRODUCTION AND OBJECTIVES: The purpose of this study was to evaluate patients diagnosed with 22q11.2 deletion syndrome and determine the clues directing to diagnosis and evaluation of immunological findings for excellent management of the disease. MATERIAL AND METHODS: Thirty-three pediatric patients with 22q11.2 deletion syndrome diag­nosed between 1998 and 2019 at Pediatric Immunology Division of Ege University Faculty of Medicine and SBU Izmir Dr Behcet Uz Children's Education and Research Hospital were evaluated. RESULTS: This study includes the largest case series reported from Turkey. Congenital car­diac anomalies were the most common pathology associated with the syndrome (90.9%). Hypocalcemic symptoms were observed in 13 patients (40%). Twenty-two of the 33 (66.6%) patients were diagnosed before two years of age. Autoimmune diseases, dysmorphic facial findings, recurrent infections, growth retardation, and speech impairment were other clues for diagnosis in older patients. Clinical spectrum and immunological abnormalities of this syn­drome are quite variable. All T-cell subset counts were less than 5th percentile below median by age in one patient (3%) and 10 patients had normal all T-cell subset counts (30.3%). Overall, 69.6% of the patients had normal IgG, IgA, and IgM levels and two patients had panhypogam­maglobulinemia. Recurrent infections were revealed in 75.7% of the patients during follow-up. CONCLUSIONS: Presence of cardiac anomaly is more helpful in the diagnosis, especially under two years of age. Patients with immunologically high or standard risk did not show any differ­ence in terms of numbers and severity of infections and autoimmunity


No disponible


Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Síndrome da Deleção 22q11/diagnóstico , Síndromes de Imunodeficiência/epidemiologia , Síndromes de Imunodeficiência/imunologia , Transtornos Cromossômicos/epidemiologia , Cromossomos Humanos Par 22 , Síndrome da Deleção 22q11/imunologia , Doenças do Sistema Imunitário/diagnóstico , Doenças do Sistema Imunitário/imunologia , Testes Imunológicos , Técnicas Imunológicas/métodos , Doenças Autoimunes/diagnóstico , Doenças Autoimunes/imunologia
7.
Clin Microbiol Infect ; 27(4): 636.e1-636.e4, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33421573

RESUMO

OBJECTIVES: There is limited information on the performance of rapid antigen detection (RAD) tests to identify SARS-CoV-2-infected asymptomatic individuals. In this field study, we evaluated the Panbio™ COVID-19 Ag Rapid Test Device (Abbott Diagnostics, Jena, Germany) for this purpose. METHODS: A total of 634 individuals (355 female; median age, 37 years; range, 9-87) were enrolled. Two nasopharyngeal swabs were collected from household (n = 338) and non-household contacts (n = 296) of COVID-19 cases. RAD testing was carried out at the point of care. The RT-PCR test used was the TaqPath COVID-19 Combo Kit (Thermo Fisher Scientific, MA, USA). RESULTS: Household contacts were tested at a median of 2 days (range, 1-7) after diagnosis of the index case, whereas non-household contacts (n = 296) were tested at a median of 6 days (range, 1-7) after exposure. In total, 79 individuals (12.4%) tested positive by RT-PCR, of whom 38 (48.1%) yielded positive RAD results. The overall sensitivity and specificity of the RAD test was 48.1% (95% CI 37.4-58.9) and 100% (95% CI 99.3-100), respectively. Sensitivity was higher in household (50.8%; 95% CI 38.9-62.5) than in non-household (35.7%; 95% CI 16.3-61.2%) contacts. Individuals testing positive by RAD test were more likely (p < 0.001) to become symptomatic than their negative counterparts. DISCUSSION: The Panbio test displays low sensitivity in asymptomatic close contacts of COVID-19 patients, particularly in non-household contacts. Nonetheless, establishing the optimal timing for upper respiratory tract collection in this group seems imperative to pinpoint test sensitivity.


Assuntos
Infecções Assintomáticas , /diagnóstico , Testes Imunológicos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos Virais/análise , Criança , Busca de Comunicante , Características da Família , Feminino , Humanos , Imunoensaio , Masculino , Pessoa de Meia-Idade , Sistemas Automatizados de Assistência Junto ao Leito , Sensibilidade e Especificidade , Adulto Jovem
8.
J Med Virol ; 93(4): 2523-2528, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33463719

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has proven to be extremely contagious and has spread rapidly all over the world. A key aspect in limiting the virus diffusion is to ensure early and accurate diagnosis. Serological assays could be an alternative in increasing testing capabilities, particularly when used as part of an algorithmic approach combined with molecular analysis. The aim of this study was to evaluate the diagnostic accuracy of a second generation chemiluminescent automated immunoassay able to detect anti-SARS-CoV-2 immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies. Data are carried out on healthy subjects and other infectious diseases pre-pandemic sera, as controls, and on two different coronavirus disease 2019 hospitalized patient groups (early and late infection time). Data obtained have been analyzed in terms of precision, linearity, sensitivity and specificity. Specificities are: 100% for anti-SARS-CoV-2 IgG and 98% for anti-SARS-CoV-2 IgM, in all patient groups. Sensitivities are: 97%, 100%, and 98% for anti-SARS-CoV-2 IgG and 87%, 83%, and 86% for anti-SARS-CoV-2 IgM in the early infection, in the late infection and in the total patient group, respectively. The Mindray anti-SARS-CoV-2 IgG and IgM assays demonstrated higher sensitivity and specificity, indicating that IgG and IgM simultaneous detection is useful even in the early phases of infection.


Assuntos
Anticorpos Antivirais/sangue , /sangue , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antivirais/imunologia , /imunologia , Reações Cruzadas , Feminino , Humanos , Imunoensaio/métodos , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Testes Imunológicos , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Glicoproteína da Espícula de Coronavírus/imunologia
9.
Int J Mol Sci ; 22(3)2021 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-33498248

RESUMO

Hereditary factor XIII (FXIII) deficiency is a rare autosomal bleeding disorder which can cause life-threatening bleeding. Acquired deficiency can be immune-mediated or due to increased consumption or reduced synthesis. The most commonly used screening test is insensitive, and widely used quantitative assays have analytical limitations. The present study sought to validate Technofluor FXIII Activity, the first isopeptidase-based assay available on a routine coagulation analyser, the Ceveron s100. Linearity was evidenced throughout the measuring range, with correlation coefficients of >0.99, and coefficients of variation for repeatability and reproducibility were <5% and <10%, respectively. A normally distributed reference range of 47.0-135.5 IU/dL was derived from 154 normal donors. Clinical samples with Technofluor FXIII Activity results between 0 and 167.0 IU/dL were assayed with Berichrom® FXIII Activity, a functional ammonia release assay, and the HemosIL™ FXIII antigen assay, generating correlations of 0.950 and 0.980, respectively. Experiments with a transglutaminase inhibitor showed that Technofluor FXIII Activity can detect inhibition of enzymatic activity. No interference was exhibited by high levels of haemolysis and lipaemia, and interference by bilirubin was evident at 18 mg/dL, a level commensurate with severe liver disease. Technofluor FXIII Activity is a rapid, accurate and precise assay suitable for routine diagnostic use with fewer interferents than ammonia release FXIII activity assays.


Assuntos
Automação Laboratorial/métodos , Testes de Coagulação Sanguínea/métodos , Carbono-Nitrogênio Liases/metabolismo , Deficiência do Fator XIII/diagnóstico , Fator XIII/análise , Corantes Fluorescentes/normas , Automação Laboratorial/normas , Bilirrubina/metabolismo , Testes de Coagulação Sanguínea/normas , Compostos Cromogênicos/normas , Fator XIII/metabolismo , Deficiência do Fator XIII/sangue , Fluorometria/métodos , Fluorometria/normas , Hemólise , Humanos , Testes Imunológicos/métodos , Testes Imunológicos/normas , Reprodutibilidade dos Testes , Transglutaminases/metabolismo
10.
Cells ; 10(2)2021 01 27.
Artigo em Inglês | MEDLINE | ID: mdl-33514016

RESUMO

Monitoring antigen-specific T cell immunity relies on functional tests that require T cells and antigen presenting cells to be uncompromised. Drawing of blood, its storage and shipment from the clinical site to the test laboratory, and the subsequent isolation, cryopreservation and thawing of peripheral blood mononuclear cells (PBMCs) before the actual test is performed can introduce numerous variables that may jeopardize the results. Therefore, no T cell test is valid without assessing the functional fitness of the PBMC being utilized. This can only be accomplished through the inclusion of positive controls that actually evaluate the performance of the antigen-specific T cell and antigen presenting cell (APC) compartments. For Caucasians, CEF peptides have been commonly used to this extent. Moreover, CEF peptides only measure CD8 cell functionality. We introduce here universal CD8+ T cell positive controls without any racial bias, as well as positive controls for the CD4+ T cell and APC compartments. In summary, we offer new tools and strategies for the assessment of PBMC functional fitness required for reliable T cell immune monitoring.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Leucócitos Mononucleares/imunologia , Células Apresentadoras de Antígenos/imunologia , Antígenos/imunologia , Humanos , Testes Imunológicos/métodos , Peptídeos/imunologia
11.
Sensors (Basel) ; 21(2)2021 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-33418986

RESUMO

Rapid diagnostic tests (RDTs) are often used for the detection of anti-human immunodeficiency virus (HIV) antibodies in remote locations in low- and middle-income countries (LMIC) with low or limited access to central laboratories. The typical format of an RDT is a lateral flow assay (LFA) with visual interpretation prone to subjectivity. This risk of misinterpretation can be overcome with luminescent upconverting nanoparticle reporters (UCNPs) measured with a miniaturized easy-to-use reader instrument. An LFA with UCNPs for anti-HIV-1/2 antibodies was developed and the assay performance was evaluated extensively with challenging patient sample panels. Sensitivity (n = 145) of the UCNP-LFA was 96.6% (95% CI: 92.1-98.8%) and specificity (n = 309) was 98.7% (95% CI: 96.7-99.7%). Another set of samples (n = 200) was used for a comparison between the UCNP-LFA and a conventional visual RDT. In this comparison, the sensitivities for HIV-1 were 96.4% (95% CI: 89.8-99.3%) and 97.6% (95% CI: 91.6-99.7%), for the UCNP-LFA and conventional RDT, respectively. The specificity was 100% (95% CI: 96.4-100%) for both assays. The developed UCNP-LFA demonstrates the applicability of UCNPs for the detection of anti-HIV antibodies. The signal measurement is done by a reader instrument, which may facilitate automated result interpretation, archiving and transfer of data from de-centralized locations.


Assuntos
HIV-1 , Nanopartículas , Anticorpos , Humanos , Imunoensaio , Testes Imunológicos , Sensibilidade e Especificidade
12.
Int J Mol Sci ; 22(2)2021 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-33445768

RESUMO

Rheumatoid arthritis (RA) is a chronic systemic inflammatory disease mainly involving synovial inflammation and articular bone destruction. RA is a heterogeneous disease with diverse clinical presentations, prognoses and therapeutic responses. Following the first discovery of rheumatoid factors (RFs) 80 years ago, the identification of both anti-citrullinated protein antibodies (ACPAs) and anti-carbamylated protein antibodies (anti-CarP Abs) has greatly facilitated approaches toward RA, especially in the fields of early diagnosis and prognosis prediction of the disease. Although these antibodies share many common features and can function synergistically to promote disease progression, they differ mechanistically and have unique clinical relevance. Specifically, these three RA associating auto-antibodies (autoAbs) all precede the development of RA by years. However, while the current evidence suggests a synergic effect of RF and ACPA in predicting the development of RA and an erosive phenotype, controversies exist regarding the additive value of anti-CarP Abs. In the present review, we critically summarize the characteristics of these autoantibodies and focus on their distinct clinical applications in the early identification, clinical manifestations and prognosis prediction of RA. With the advancement of treatment options in the era of biologics, we also discuss the relevance of these autoantibodies in association with RA patient response to therapy.


Assuntos
Anticorpos Anti-Proteína Citrulinada/imunologia , Artrite Reumatoide/diagnóstico , Artrite Reumatoide/imunologia , Autoanticorpos/imunologia , Fator Reumatoide/imunologia , Autoantígenos/imunologia , Autoimunidade , Biomarcadores , Progressão da Doença , Humanos , Testes Imunológicos , Prognóstico
13.
J Infect Chemother ; 27(2): 384-386, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33397587

RESUMO

We evaluated the rapid immunochromatographic test for severe acute respiratory coronavirus 2 (SARS-CoV-2) antigen detection using 16 saliva specimens collected from 6 COVID-19 hospitalized patients, and detected N-antigen in 4 of 7 RT-PCR positive specimens. This POCT detected SARS-CoV-2 antigen in saliva and would be useful for COVID-19 diagnosis.


Assuntos
Antígenos Virais/análise , /diagnóstico , Saliva/virologia , Humanos , Testes Imunológicos , Nasofaringe/virologia , Testes Imediatos , RNA Viral/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sensibilidade e Especificidade
15.
BMC Infect Dis ; 21(1): 118, 2021 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-33499820

RESUMO

BACKGROUND: Early infant diagnosis of HIV infection is challenging in sub-Saharan Africa, particularly in rural areas, leading to delays in diagnosis and treatment. Use of a point-of-care test would overcome many challenges. This study evaluated the validity of a novel point-of-care p24 antigen detection test (LYNX) in rural and urban settings in southern Zambia. METHODS: Two studies were conducted: a cross-sectional study from 2014 to 2015 at Macha Hospital (LYNX Hospital study) and a longitudinal study from 2016 to 2018 at 12 health facilities in Southern Province, Zambia (NSEBA study). In both studies, children attending the facilities for early infant diagnosis were enrolled and a blood sample was collected for routine testing at the central lab and immediate on-site testing with the LYNX test. The performance of the LYNX test was measured in comparison to nucleic acid-based testing at the central lab. RESULTS: In the LYNX Hospital study, 210 tests were performed at a median age of 23.5 weeks (IQR: 8.9, 29.0). The sensitivity and specificity of the test were 70.0 and 100.0%, respectively. In the NSEBA study, 2608 tests were performed, including 1305 at birth and 1222 on children ≥4 weeks of age. For samples tested at birth, sensitivity was 13.6% (95% CI: 2.9, 34.9) and specificity was 99.6% (95% CI: 99.1, 99.9). While specificity was high for all ages, sensitivity increased with age and was higher for participants tested at ≥4 weeks of age (80.6%; 95% CI: 67.4, 93.7). Children with positive nucleic acid tests were more likely to be negative by the LYNX test if their mother received antiretroviral therapy during pregnancy (60.7% vs. 24.2%; p = 004). CONCLUSIONS: Considering the high specificity and moderate sensitivity that increased with age, the LYNX test could be of value for early infant diagnosis for infants ≥4 weeks of age, particularly in rural areas where centralized testing leads to long delays. Point-of-care tests with moderate sensitivity and high specificity that are affordable, easy-to-use, and easily implemented and maintained should be developed to expand access to testing and deliver same-day results to infants in areas where it is not feasible to implement nucleic acid-based point-of-care assays.


Assuntos
Proteína do Núcleo p24 do HIV/análise , Infecções por HIV/diagnóstico , Testes Imediatos , Fármacos Anti-HIV/uso terapêutico , Estudos Transversais , Testes Diagnósticos de Rotina , Diagnóstico Precoce , Estudos de Viabilidade , Feminino , Proteína do Núcleo p24 do HIV/sangue , Infecções por HIV/congênito , Infecções por HIV/epidemiologia , Infecções por HIV/transmissão , Humanos , Testes Imunológicos , Ciência da Implementação , Lactente , Recém-Nascido , Transmissão Vertical de Doença Infecciosa , Estudos Longitudinais , Masculino , Triagem Neonatal/métodos , Sistemas Automatizados de Assistência Junto ao Leito , Gravidez , Complicações Infecciosas na Gravidez/tratamento farmacológico , Complicações Infecciosas na Gravidez/epidemiologia , População Rural , Sensibilidade e Especificidade , Zâmbia/epidemiologia
16.
Methods Mol Biol ; 2237: 179-189, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33237417

RESUMO

Recent advances in biosensing analytical platforms have brought relevant outcomes for novel diagnostic and therapy-oriented applications. In this context, hydrogels have emerged as appealing matrices to locally confine biomolecules onto sensing surfaces under solution mimetic conditions, preserving their structural integrity and function. Here, we describe the application of a self-assembling peptide hydrogel as a suitable matrix for 3D microarray bioassays. The hydrogel is printable and self-adhesive and allows for fast analyte diffusion. As a showcase example, we describe its application in a diagnostic immunoassay for the detection of arbovirus infection.


Assuntos
Bioimpressão/métodos , Hidrogéis/química , Testes Imunológicos/métodos , Análise Serial de Proteínas/métodos , Animais , Infecções por Arbovirus/diagnóstico , Humanos , Imunoensaio/métodos , Peptídeos/química
17.
Methods Mol Biol ; 2237: 237-245, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33237423

RESUMO

Biomarkers for diseases are important for the development of clinical diagnostic tests and can provide early intervention for cancer or cardiovascular patients. Over the past decade, antibody array technology has achieved significant technological improvement in the quantitative measurement of more than a thousand proteins simultaneously and has been utilized to screen and identify unique proteins as disease biomarkers. However, few biomarkers have been translated into clinical application. This chapter will discuss the protocol for the screening and validation of unique proteins that create a new avenue for biomarker discovery.


Assuntos
Análise Serial de Proteínas/métodos , Proteômica/métodos , Biomarcadores/sangue , Bioimpressão/métodos , Humanos , Imunoensaio/métodos , Testes Imunológicos/métodos , Aprendizado de Máquina
18.
Methods Mol Biol ; 2237: 1-10, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33237404

RESUMO

Sandwich-based antibody arrays enable the detection of multiple proteins simultaneously, thus offering a time- and cost-effective alternative to single-plex platforms. The protein of interest is "sandwiched" between an antibody that captures it to the array and a second antibody that is used for detection. Here we describe a 1-day procedure to process samples, such as serum or cell lysates, with a quantitative sandwich-based antibody array on a glass substrate using fluorescence.


Assuntos
Imunofluorescência/métodos , Testes Imunológicos/métodos , Análise Serial de Proteínas/métodos , Animais , Anticorpos/imunologia , Citocinas/análise , Citocinas/imunologia , Humanos
19.
Methods Mol Biol ; 2237: 11-38, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33237405

RESUMO

Multiplex immunoassays are important tools in basic research and diagnostics. The ability to accurately quantify the presence of several antigens within an individual sample all at once has been useful in developing a proteomics view of biology. This in turn has enabled the development of disease-associated immunodiagnostic panels for better prognosis and well-being. Moreover, it is well understood that such multiplexing approaches lend themselves to automation, thereby reducing labor while providing the ability to dramatically conserve both reagent and sample all of which will reduce the cost per test. Here we describe various methods to create and use multiplex immunoassays in the wells of microtiter plates or similar formats.


Assuntos
Testes Imunológicos/métodos , Análise Serial de Proteínas/métodos , Animais , Automação Laboratorial/métodos , Automação Laboratorial/normas , Humanos , Técnicas Imunoenzimáticas/métodos , Técnicas Imunoenzimáticas/normas , Testes Imunológicos/economia , Testes Imunológicos/normas , Análise Serial de Proteínas/normas , Sensibilidade e Especificidade
20.
Methods Mol Biol ; 2237: 39-44, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33237406

RESUMO

The sandwich-based immunoassay is renowned for its specificity and sensitivity for protein detection, where the antigen is "sandwiched" by a pair of antigen epitope-specific antibodies. The capture antibody-antigen-detection antibody complex will be formed if all the components are present in a proper reaction system. A one-step rapid sandwich-based antibody array can be developed through fixing the capture antibody on a glass slide with a fluorescence-labelled detection antibody. In this chapter, we describe the process of a one-step mouse immunoglobulin isotyping array for use in hybridoma culture supernatants.


Assuntos
Testes Imunológicos/métodos , Análise Serial de Proteínas/métodos , Animais , Imunofluorescência/métodos , Humanos , Imunoglobulinas/imunologia
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