Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 1.700.971
Filtrar
1.
Methods Mol Biol ; 2578: 1-15, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36152276

RESUMO

The diversity of the antigen-specific humoral immune response reflects the interaction of the immune system with pathogens and autoantigens. Peptide microarray analysis opens up new perspectives for the use of antibodies as diagnostic biomarkers and provides unique access to a more differentiated view on humoral responses to disease. This review focuses on the latest applications of peptide microarrays for the serologic medical diagnosis of autoimmunity, infectious diseases (including COVID-19), and cancer.


Assuntos
COVID-19 , Neoplasias , Autoanticorpos/análise , Autoantígenos , Biomarcadores , COVID-19/diagnóstico , Humanos , Análise em Microsséries , Neoplasias/diagnóstico , Peptídeos , Análise Serial de Proteínas
2.
Methods Mol Biol ; 2578: 17-25, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36152277

RESUMO

Antibody-mediated neurological diseases constitute an emerging clinical entity that remains to be fully explored. Recent studies identified autoantibodies that directly confer pathogenicity, and it was shown that in these cases immunotherapies can result in profound positive patient responses. These advances highlight the urgent need for improved means to effectively screen patient samples for novel autoantibodies (aAbs) and their subsequent characterization. Here, we discuss challenges and opportunities for peptide microarrays to contribute to the identification, mapping, and characterization of the underlying monospecific disease-defining binding surfaces. We outline control experiments, workflow modifications and bioinformatic filtering methods that enhance the predictive power of array-based studies. Further, we highlight experimental and computer-based display approaches that have the potential to expand the use of synthetic microarrays over the detection of discontinuous epitopes. Knowledge over the autoantibody epitopes in neurological disease will enhance our understanding of the pathological mechanisms and thereby potentially contribute to novel diagnostic approaches or even innovative antigen-specific treatments that avoid the serious adverse effects seen with currently used immunosuppressive therapies.


Assuntos
Autoanticorpos , Doenças do Sistema Nervoso , Biologia Computacional , Mapeamento de Epitopos/métodos , Epitopos , Humanos , Análise em Microsséries , Doenças do Sistema Nervoso/diagnóstico , Peptídeos/química
3.
Methods Mol Biol ; 2578: 53-62, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36152280

RESUMO

Recent advances in biosensing analytical platforms have brought relevant outcomes for novel diagnostic and therapy-oriented applications. In this context, 3D droplet microarrays, where hydrogels are used as matrices to stably entrap biomolecules onto analytical surfaces, potentially provide relevant advantages over conventional 2D assays, such as increased loading capacity, lower nonspecific binding, and enhanced signal-to-noise ratio. Here, we describe a hybrid hydrogel composed of a self-assembling peptide and commercial agarose (AG) as a suitable matrix for 3D microarray bioassays. The hybrid hydrogel is printable and self-adhesive and allows analyte diffusion. As a showcase example, we describe its application in a diagnostic immunoassay for the detection of SARS-CoV-2 infection.


Assuntos
COVID-19 , Hidrogéis , COVID-19/diagnóstico , Humanos , Hidrogéis/química , Imunoensaio , Peptídeos/química , Cimentos de Resina , SARS-CoV-2 , Sefarose
4.
Methods Mol Biol ; 2578: 209-217, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36152290

RESUMO

In SARS-CoV-2 pandemic scenario, the identification of rapid methods to detect antibodies against coronavirus has been a wide and urgent issue. Epitope mapping on peptide microarrays is a rapid way to identify sequences with a high immunoreactivity. The process begins with a proteome-wide screening, based on immune affinity; the use of a high-density microarray is followed by a validation phase, where a restricted panel of probes is tested using peptide microarrays; peptide sequences are immobilized through a click-based strategy.COVID-19-positive sera are tested and immuno-domains regions are identified on SARS-CoV-2 spike (S), nucleocapsid (N) protein, and Orf1ab polyprotein. An epitope on N protein (region 155-171) provided good diagnostic performance in discriminating COVID-19-positive vs. healthy individuals. Using this sequence, 92% sensitivity and 100% specificity are reached for IgG detection in COVID-19 samples, and no cross-reactivity with common cold coronaviruses is detected. Overall, epitope 155-171 from N protein represents a promising candidate for further development and rapid implementation in serological tests.


Assuntos
COVID-19 , SARS-CoV-2 , Anticorpos Antivirais , COVID-19/diagnóstico , Mapeamento de Epitopos , Epitopos , Humanos , Imunidade , Imunoglobulina G , Poliproteínas , Proteoma , SARS-CoV-2/genética , Glicoproteína da Espícula de Coronavírus
5.
Methods Mol Biol ; 2578: 219-236, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36152291

RESUMO

Peptide microarrays are a powerful tool to identify linear epitopes of food allergens in a high-throughput manner. The main advantages of the microarray-based immunoassay are as follows: the possibility to assay thousands of targets simultaneously, the requirement of a low volume of serum, the more robust statistical analysis, and the possibility to test simultaneously several immunoglobulin subclasses. Among them, the last one has a special interest in the field of food allergy, because the development of tolerance to food allergens has been associated with a decrease in IgE and an increase in IgG4 levels against linear epitopes. However, the main limitation to the clinical use of microarray is the automated analysis of the data. Recent studies mapping the linear epitopes of food allergens with peptide microarray immunoassays have identified peptide biomarkers that can be used for early diagnosis of food allergies and to predict their severity or the self-development of tolerance. Using this approach, we have worked on epitope mapping of the two most important food allergens in the Spanish population, cow's milk, and chicken eggs. The final aim of these studies is to define subsets of peptides that could be used as biomarkers to improve the diagnosis and prognosis of food allergies. This chapter describes the protocol to produce microarrays using a library of overlapping peptides corresponding to the primary sequences of food allergens and data acquisition and analysis of IgE and IgG4 binding epitopes.


Assuntos
Hipersensibilidade Alimentar , Imunoglobulina G , Alérgenos , Animais , Biomarcadores , Bovinos , Mapeamento de Epitopos/métodos , Epitopos , Feminino , Hipersensibilidade Alimentar/diagnóstico , Imunoensaio/métodos , Imunoglobulina E/metabolismo , Peptídeos
6.
Adv Neurobiol ; 29: 219-253, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36255677

RESUMO

The chapter is devoted to neurological aspects of congenital disorders of glycosylation (CDG). At the beginning, the various types of CDG with neurological presentation of symptoms are summarized. Then, the occurrence of various neurological constellation of abnormalities (for example: epilepsy, brain anomalies on neuroimaging, ataxia, stroke-like episodes, autistic features) in different CDG types are discussed followed by data on possible biomarkers and limited treatment options.


Assuntos
Defeitos Congênitos da Glicosilação , Epilepsia , Humanos , Defeitos Congênitos da Glicosilação/diagnóstico , Glicosilação , Biomarcadores
7.
Talanta ; 251: 123759, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-35952499

RESUMO

Heart failure (HF) is a chronic cardiovascular disease that represents main cause of mortality worldwide, particularly for elderly. N-terminal pro-brain natriuretic peptide (NT-proBNP) was identified as the gold standard biomarker for HF diagnosis and therapy monitoring. Presently, saliva analysis represents an emerging and powerful tool for clinical applications and electrochemical immunosensors have shown their potential in Healthcare applications as selective and reliable systems for detecting clinical biomarkers. This work presents the detection of NT-proBNP in saliva samples by an immunologically modified Field effect Transistor (IMFET). TESUD ((11-triethoxysilyl) undecanal) was used as cross-linker to immobilise anti-NT-proBNP antibody onto the device. Our IMFET that was then tested in different matrices (e.g. phosphate buffered saline (PBS), artificial saliva and human saliva) using electrochemical impedance spectroscopy (EIS), and it resulted selective to NT-proBNP with good sensitivity (detection limit of 0.02 pg/mL) and a wide linear range (0.02-1 pg/mL and 0.5-20 pg/mL). Finally, NT-proBNP concentration in ten saliva samples was determined by performing the standard addition method. An enzyme-linked immunosorbent assay was used for confirming IMFET results, highlighting both IMFET accuracy (analyte recovery of 99 ± 8%) and precision (coefficient of variation always <10%), and supporting the suitability of the device for determining salivary NT-proBNP.


Assuntos
Técnicas Biossensoriais , Insuficiência Cardíaca , Idoso , Humanos , Biomarcadores , Insuficiência Cardíaca/diagnóstico , Imunoensaio , Peptídeo Natriurético Encefálico , Fragmentos de Peptídeos , Fosfatos , Saliva , Saliva Artificial , Volume Sistólico , Técnicas Eletroquímicas
8.
Food Chem ; 398: 133884, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-35964575

RESUMO

Development of selective and sensitive methods for the detection of 2, 6-dipicolinic acid (DPA), a biomarker produced by bacterial spores, is of great significance for maintaining public health and food safety. Herein, a ratiometric fluorescence strategy using graphene carbon nitride (g-C3N4) coupled with Eu3+ is designed for the assay of DPA. As the concentration of DPA increases, the emission intensity of g-C3N4 kept constant which acted as a stable internal reference, while the fluorescence of Eu3+ was enhanced obviously due to the antenna effect. The linear calibration ranged from 0.1 to 15 µM with a detection limit of 13 nM was obtained. More Importantly, a paper-based sensor with a smartphone was successfully combined to perform colorimetric and visual detection of DPA in situ. This method has good performance for the detection of DPA, which is expected to broaden the application prospects of preliminary biomarker monitoring.


Assuntos
Antraz , Antraz/diagnóstico , Antraz/microbiologia , Biomarcadores , Európio , Corantes Fluorescentes , Humanos , Nitrilas , Ácidos Picolínicos , Smartphone
10.
Ann Lab Med ; 43(1): 38-44, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36045055

RESUMO

Background: Reference materials are essential for the quality assurance of molecular detection methods. We developed and characterized synthetic norovirus GI and GII RNA reference materials. Methods: Norovirus GI and GII RNA sequences including the ORF1-ORF2 junction region were designed based on 1,495 reported norovirus sequences and synthesized via plasmid preparation and in vitro transcription. The synthetic norovirus GI and GII RNAs were evaluated using six commercial norovirus detection kits used in Korea and subjected to homogeneity and stability analyses. A multicenter study involving five laboratories and using four commercial real-time PCR norovirus detection assays was conducted for synthetic norovirus RNA characterization and uncertainty measurements. Results: The synthetic norovirus GI and GII RNAs were positively detected using the six commercial norovirus detection kits and were homogeneous and stable for one year when stored at -20°C or -70°C. All data from the five laboratories were within a range of 1.0 log copies/µL difference for each RNA, and the overall mean concentrations for norovirus GI and GII RNAs were 7.90 log copies/µL and 6.96 log copies/µL, respectively. Conclusions: The synthetic norovirus GI and GII RNAs are adequate for quality control based on commercial molecular detection reagents for noroviruses with high sequence variability. The synthetic RNAs can be used as reference materials in norovirus molecular detection methods.


Assuntos
Infecções por Caliciviridae , Norovirus , Infecções por Caliciviridae/diagnóstico , Genótipo , Humanos , Norovirus/genética , RNA Viral/análise , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , República da Coreia
11.
Ann Lab Med ; 43(1): 64-72, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36045058

RESUMO

Background: Following success of the phase III PROfound trial, the poly (ADP-ribose) polymerase (PARP) inhibitor olaparib was approved by the US Food and Drug Administration in May 2020 for adult patients with deleterious homologous recombination repair (HRR) gene-mutated metastatic castration-resistant prostate cancer (mCRPC). As locally adopted multigene panel next-generation sequencing (NGS) assays for selecting PARP inhibitor candidates have not been thoroughly evaluated, we compared the analytical performance of the FoundationOne CDx (Foundation Medicine, Inc., Cambridge, MA, USA) (central laboratory) and other NGS assays (local laboratory) with samples from the PROfound trial in Korea. Methods: One hundred PROfound samples (60 HRR mutation [HRRm] cases and 40 non-HRRm cases) were analyzed. The results of HRR gene mutation analysis were compared between the FoundationOne CDx and two other NGS assays [SureSelect Custom Design assay (Agilent Technologies, Inc., Santa Clara, CA, USA) and Oncomine Comprehensive assay (Thermo Fisher Scientific, Inc., Waltham, MA, USA)]. Results: The positive percent agreement for single nucleotide variants (SNVs) and insertion/deletions (indels) between the central laboratory and local laboratory was 98.7%-100.0%. The negative percent agreement and overall percent agreement (OPA) for SNVs and indels between central and local laboratories were both 100%. Compared with that of the FoundationOne CDx assay, the OPA for copy number variations of the Oncomine Comprehensive and SureSelect Custom assays reached 99.8%-100%. Most mCRPC patients harboring a deleterious genetic variant were successfully identified with both local laboratory assays. Conclusions: The NGS approach at a local laboratory showed comparable analytical performance for identifying HRRm status to the FoundationOne CDx assay used at the central laboratory.


Assuntos
Neoplasias de Próstata Resistentes à Castração , Adulto , Variações do Número de Cópias de DNA , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Laboratórios , Masculino , Neoplasias de Próstata Resistentes à Castração/diagnóstico , Neoplasias de Próstata Resistentes à Castração/tratamento farmacológico , Neoplasias de Próstata Resistentes à Castração/genética , Reparo de DNA por Recombinação
12.
Ann Lab Med ; 43(1): 73-81, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36045059

RESUMO

Background: BRCA testing is necessary for establishing a management strategy for ovarian cancer. Several BRCA testing strategies, including germline and somatic testing, are implemented in clinical practice in Korea. We aimed to comparatively evaluate their cost-effectiveness from patients' perspective. Methods: We developed a decision model comprising five BRCA testing strategies implemented in Korea: (1) germline testing first, followed by somatic tumor testing for patients without a germline variant; (2) somatic testing first, followed by germline testing for patients with a variant detected by somatic testing; (3) both germline and somatic testing; (4) germline testing alone; and (5) somatic testing alone, with no testing as the comparator. One-way sensitivity analysis was conducted to test the uncertainty of key parameters. Results: Assuming a willingness-to-pay of $20,000 per progression-free life-year gain (PF-LYG), all five strategies were considered cost-effective. Strategy 4 was the most cost-effective option, with an incremental cost-effectiveness ratio (ICER) of $2,547.7 per PF-LYG, followed by strategy 1, with an ICER of $3,978.4 per PF-LYG. Even when the parameter values were varied within the possible range, the ICERs of all strategies did not exceed the willingness-to-pay threshold. Conclusions: Considering the importance of knowing a patient's BRCA gene status, germline testing first, followed by somatic testing, may be a reasonable option.


Assuntos
Neoplasias Ovarianas , Análise Custo-Benefício , Feminino , Células Germinativas/patologia , Humanos , Neoplasias Ovarianas/diagnóstico , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , República da Coreia
13.
Ann Lab Med ; 43(1): 82-85, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36045060

RESUMO

Cytokeratin 19 fragment antigen 21-1 (CYFRA 21-1) is useful for predicting and monitoring non-small cell lung cancer prognosis. We established reference intervals (RIs) of CYFRA 21-1 in Korean adults, including those older than 60 years. Data of 4,098 apparently healthy subjects (age range, 20-87 years) were analyzed after excluding those with a history of malignancy, high tumor marker concentrations (except CYFRA 21-1), and/or abnormal findings on a chest computed tomography scan through medical chart review. After removing two outliers, RIs of CYFRA 21-1 were determined using data of 4,096 subjects based on the non-parametric method (2.5th and 97.5th percentiles) according to CLSI guidelines EP28-A3c. The subjects were divided into two and four groups according to sex and age (20-40, 41-50, 51-60, and >60 years), respectively, and the median CYFRA 21-1 concentration was compared between the groups. The RI of CYFRA 21-1 was 0.66-3.84 ng/mL, applicable to both men and women. Regardless of sex, the CYFRA 21-1 concentration increased with age, suggesting that age-dependent RIs of CYFRA 21-1 should be applied. Rather than using a single RI provided by the manufacturer, the RI of CYFRA 21-1 should be continually verified and established in each clinical laboratory.


Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos de Neoplasias , Biomarcadores Tumorais , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Carcinoma Pulmonar de Células não Pequenas/patologia , Feminino , Humanos , Queratina-19 , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , República da Coreia , Adulto Jovem
14.
Ann Lab Med ; 43(1): 86-91, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36045061

RESUMO

Antibodies against human CD36 are responsible for several immune-mediated disorders. The detection of anti-CD36 antibodies using the standard monoclonal antibody (mAb) immobilization of platelet antigens (MAIPA) assay is hampered by a high frequency of false-negative results, most likely due to competitive inhibition of the mAb used as the capture antibody. We generated a panel of mouse mAbs against CD36 and seven hybridomas (GZ-3, GZ-13, GZ-70, GZ-143, GZ-413, GZ-507, and GZ-608), which were selected for MAIPA assays, as they reacted with mouse and human CD36. Fourteen anti-CD36 sera were assayed; all of which showed a positive reaction in a PakPlus (Immucor GTI Diagnostics, Inc., Waukesha, WI, USA) ELISA-based screening (optical density: 0.257-2.292). When the reference anti-CD36 mAb FA6-152 was used in the MAIPA assay, only 6/14 (42.9%) sera displayed a positive reaction. In contrast, anti-CD36 antibodies were detected in 13/14 (92.9%) sera when GZ-70 and GZ-608 mAbs were used. This significant improvement resulted in the identification of anti-CD36 antibodies by an antigen capture assay. Since patient's platelets possibly carrying rare native antigens are used, this method will facilitate the identification of new platelet antibodies against CD36 that are involved in immune-mediated thrombocytopenia and other diseases, such as transfusion-related acute lung injury.


Assuntos
Antígenos de Plaquetas Humanas , Trombocitopenia , Animais , Anticorpos Monoclonais , Plaquetas , Antígenos CD36 , Humanos , Camundongos , Trombocitopenia/diagnóstico
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...