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1.
Methods Mol Biol ; 2597: 121-129, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36374418

RESUMO

Viruses encode secreted proteins that bind chemokines to modulate their activity. Viral proteins may simultaneously interact with glycosaminoglycans allowing these proteins to be anchored at the cell surface to increase their anti-chemokine activity in the proximity of infection. Here we describe methodology to evaluate the interaction of viral secreted proteins with cell-surface glycosaminoglycans by immunofluorescence and detection by flow cytometry or microscopy. These methods could be equally applied to other chemokine binding proteins that do not have viral origin.


Assuntos
Proteínas de Transporte , Glicosaminoglicanos , Glicosaminoglicanos/metabolismo , Proteínas de Transporte/metabolismo , Técnica Indireta de Fluorescência para Anticorpo , Citometria de Fluxo , Quimiocinas/metabolismo , Ligação Proteica , Proteínas Virais/metabolismo
2.
Klin Lab Diagn ; 67(11): 652-657, 2022 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-36398774

RESUMO

Antinuclear antibodies (ANA) are a heterogeneous group of autoantibodies that react with various components of the cell nucleus and cytoplasm. ANA is the main serological marker for autoimmune liver disease (AILD). The aim of the study was to compare the diagnostic value of two methods of screening for the determination of ANA (indirect immunofluorescence reaction on HEp-2 cells (IIF -HEp-2) and enzyme-linked immunosorbent assay (ELISA) in the sera of AILD patients. The sera of 118 patients with AILD (51 with autoimmune hepatitis - AIH, 19 with primary biliary cholangitis - PBC, 48 with overlapping syndrome - OVERLAP), 30 patients with non-alcoholic fatty liver disease (NAFLD) and 30 healthy donors (HD) were studied. Determination of ANA by the IIF-HEp-2 method was carried out by visual assessment of samples under an AXIOSKOP 40 microscope, by ELISA - on an Alegria automatic analyzer. A weak degree of agreement between the positive and negative results of the ANA screening study using IIF-HEp-2 and ELISA (Cohen's kappa coefficient æ=0.4) was noted. Screening determination of ANA in patients with AILD by the IIF-HEp-2 method was distinguished by greater diagnostic sensitivity (DS) (68.6%) and a lower frequency of false negative results (31.4%) compared with ELISA (35.6% and 64.4 % respectively, p<0.05). The overall diagnostic specificity (DS) of the ANA study in IIF-HEp-2 was lower than with ELISA (66.7% and 86.7%, respectively, p<0.05). Both screening methods for determining ANA (IIF-HEp-2 and ELISA) were useful for diagnosing AILD (positive likelihood ratio - LR+: 2.1 and 2.6, respectively). In terms of the negative likelihood ratio (LR-), screening for ANA by the IIF-HEp-2 method, in contrast to ELISA, served as a "useful" test to exclude the diagnosis of AILD (0.5 and 0.8, respectively). The determination of ANA using IIF-HEp-2 is the most sensitive and "useful" screening test for the diagnosis of AILD, and ELISA is classified as a less "useful" screening method due to low diagnostic sensitivity and a high false-negative rate.


Assuntos
Doenças Autoimunes , Hepatopatias , Humanos , Técnica Indireta de Fluorescência para Anticorpo/métodos , Anticorpos Antinucleares , Doenças Autoimunes/diagnóstico , Ensaio de Imunoadsorção Enzimática
3.
Diagn Microbiol Infect Dis ; 104(4): 115809, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36244126

RESUMO

The utility of IgA class antibodies for the serodiagnosis of cat scratch disease (CSD) was evaluated by developing an indirect immunofluorescent assay (IFA) using an antigen obtained by co-cultivating Bartonella henselae ATCC 49882 with Vero cells. Served for evaluation were 101 sera from patients serologically confirmed as CSD with IgG-IFA ≥1:256, and 144 sera from patients clinically suspected of CSD but not serologically confirmed. The sensitivity of the newly developed IgA-IFA in detecting the confirmed cases was 57.4% (58/101), and 75.0% in combination with IgM-IFA. As for the non-confirmed cases, IgA-IFA turned 8.3% cases (12/144) positive, 10 of whom were subsequently diagnosed as CSD of early stage from clinical courses and/or by repeated testing. The 12-case gain was regarded as a significant improvement. Hence, the diagnostic rate of early-stage CSD is expected to be increased by routinely performing IgA-IFA in addition to conventional IgG/IgM-IFA.


Assuntos
Bartonella henselae , Doença da Arranhadura de Gato , Chlorocebus aethiops , Animais , Doença da Arranhadura de Gato/diagnóstico , Imunoglobulina A , Células Vero , Anticorpos Antibacterianos , Testes Sorológicos , Técnica Indireta de Fluorescência para Anticorpo , Imunoglobulina M , Imunoglobulina G
4.
Front Immunol ; 13: 1011110, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36225928

RESUMO

Anti-fibrillarin autoantibodies are useful for the diagnosis and prognosis of systemic sclerosis (SSc). Anti-fibrillarin produces a clumpy nucleolar pattern in indirect immunofluorescence assay on HEp-2 cells (HEp-2 IFA). Here we develop and validate a reliable cell-based anti-fibrillarin assay (Fibrillarin/CBA) for use in clinical diagnostic laboratories. A TransMembrane Signal was fused to the human fibrillarin gene (TMS-fibrillarin). HEp-2 cells overexpressing transgenic TMS-fibrillarin at the cytoplasmic membrane were used as IFA substrate in the Fibrillarin/CBA. Sixty-two serum samples with nucleolar pattern in the HEp-2 IFA (41 clumpy; 21 homogeneous/punctate) were tested for anti-fibrillarin using Fibrillarin/CBA, immunoprecipitation (IP), line-blot and ELISA. In addition, samples from 106 SSc-patients were evaluated with Fibrillarin/CBA and the results were correlated with disease phenotypes. Thirty-eight of 41 samples with the clumpy nucleolar pattern (92.7%) were positive in the Fibrillarin/CBA, while all 21 samples with other nucleolar patterns were negative. Fibrillarin/CBA results agreed 100% with IP results. Among the 38 Fibrillarin/CBA-positive samples, only 15 (39.5%) and 11 (29%) were positive for anti-fibrillarin in line-blot and ELISA, respectively. Higher frequency of diffuse cutaneous SSc (dcSSc) phenotype (72.7% vs 36.8%; p=0.022), cardiac involvement (36.4% vs 6.5%; p=0.001) and scleroderma renal crisis (18.2% vs 3.3% p = 0.028) was observed in SSc patients with positive compared to negative Fibrillarin/CBA result. Performance of Fibrillarin/CBA in the detection of anti-fibrillarin autoantibodies was comparable to the gold standard IP. Positive Fibrillarin/CBA results correlated with disease phenotypes known to be associated with anti-fibrillarin autoantibodies, underscoring the clinical validation of this novel assay.


Assuntos
Autoanticorpos , Escleroderma Sistêmico , Ensaio de Imunoadsorção Enzimática , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Imunoprecipitação , Escleroderma Sistêmico/diagnóstico
5.
Front Immunol ; 13: 1007257, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36238277

RESUMO

Objective: To investigate the distribution and clinical significance of the rods and rings (RR) pattern in various diseases. Methods: A total of 169,891 patients in Peking Union Medical College Hospital (PUMCH) and 29,458 patients in Inner Mongolia People's Hospital (IMPH) from January 2018 to December 2020 were included, and the results of ANA (antinuclear antibodies) and special antibodies were analyzed retrospectively. Results: The positive rates of ANA and RR patterns were 34.84%, 0.16% in PUMCH, and 44.73%, 0.23% in IMPH. Anti-RR antibodies mainly appear in adults (≥ 41 years), mostly of low or medium fluorescence titers. Isolated RR patterns were mostly presented (60.30% and 69.12%, respectively), and the RR pattern mixed with the speckled pattern was most commonly observed among patients having two or more patterns. The RR pattern existed in a variety of diseases including hepatitis C, AIDs, pulmonary diseases, nephropathy diseases, and even healthy people. The highest prevalence of the RR pattern was observed in hepatic diseases, such as hepatic dysfunction (0.79%), hepatic cirrhosis (1.05%), PBC (0.85%), and AIH (0.65%), etc. The positive rate of specific antibodies in RR pattern cases was 31.25%, and anti-Ro52 (27, 20.61%) was the most common target antibody. Conclusion: The RR pattern had a low prevalence in ANAs test samples and varied in different nationalities and regions. Except for hepatitis C, it could be observed in AIDs, pulmonary diseases, nephropathy, other hepatic diseases, and even healthy people, but the positive rate was slightly higher in hepatic diseases. Its mechanism of action and clinical relevance still need clarification.


Assuntos
Síndrome de Imunodeficiência Adquirida , Hepatite C , Hepatopatias , Pneumopatias , Adulto , Anticorpos Antinucleares , China/epidemiologia , Técnica Indireta de Fluorescência para Anticorpo , Hepacivirus , Humanos , Estudos Retrospectivos
6.
Indian J Pathol Microbiol ; 65(4): 873-878, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36308197

RESUMO

Background: For the management of connective tissue disorders (CTDs), antinuclear antibody (ANA) testing is essential, both from diagnostic and prognostic points of view. Usually, patterns obtained by ANA-IIF testing correlates to specific autoantibodies as obtained from the test for ENA (by LIA/ELISA, etc.). But to apply these data from western studies, we may need validation in the local population like our subjects in sub-Himalayan (Garhwal region) area where CTDs are common. Also, suppose ANA-IFA pattern's correlation is reliably known in our population, it can minimize the cost of managing CTDs by limiting ENA testing, which is 10 times costlier than ANA-IIF. Hence, this study was undertaken to know the specific autoantibody targets (ENA by LIA) against ANA-IIF patterns in our local population. Materials and Methods: In this retrospective cross-sectional work, serum samples of CTDs were tested for ANA by IIF (Euroimmune AG) and ENA by LIA (Euroline ANA-3G) continuously for 36 months. The manufacturer's kit insert was followed, and results were analyzed applying appropriate statistical methods. Results: Major ANA-IIF patterns were found to be associated with specific autoantibodies, for example, Nuclear homogenous with dsDNA, nucleosomes, histones; speckled pattern with nRNP/Sm, Sm, SSA/Ro-52, SSB; nucleolar pattern with Scl-70, Pm-Scl 100 and centromere pattern with CENP-B. Anticytoplasmic (ACA) are found to be linked with some ANA negative (by IIF) samples, emphasizing the need for careful observation for ACA especially where ANA is not found. Conclusions: In most subjects, specific ENA targets correlated well with ANA-IIF patterns, implying effective cost minimization in CTD management. Similar future prospective studies (with clinical data) can provide a database and reference for our population.


Assuntos
Anticorpos Antinucleares , Doenças do Tecido Conjuntivo , Humanos , Técnica Indireta de Fluorescência para Anticorpo/métodos , Anticorpos Antinucleares/análise , Estudos Transversais , Estudos Retrospectivos , Estudos Prospectivos , Estudos de Viabilidade , Autoanticorpos , Doenças do Tecido Conjuntivo/diagnóstico , Índia
9.
J Vet Sci ; 23(5): e32, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36174976

RESUMO

BACKGROUND: Classical swine fever (CSF) is a severe infectious disease of pigs that causes significant economic losses to the swine industry. OBJECTIVES: This study developed a solid-phase blocking enzyme-linked immunosorbent assay (spbELISA) method for the specific detection of antibodies against the CSF virus (CSFV) in porcine serum samples. METHODS: A spbELISA method was developed based on the recombinant E2 expressed in Escherichia coli. The specificity of this established spbELISA method was evaluated using reference serum samples positive for antibodies against other common infectious diseases. The stability and sensitivity were evaluated using an accelerated thermostability test. RESULTS: The spbELISA successfully detected the antibody levels in swine vaccinated with the C-strain of CSFV. In addition, the detection ability of spbELISA for CSFV antibodies was compared with that of other commercial ELISA kits and validated using an indirect immunofluorescence assay. The results suggested that the spbELISA provides an alternative, stable, and rapid serological detection method suitable for the large-scale screening of CSFV serum antibodies. CONCLUSIONS: The spbELISA has practical applications in assessing the vaccination status of large pig herds.


Assuntos
Vírus da Febre Suína Clássica , Peste Suína Clássica , Doenças dos Suínos , Animais , Anticorpos , Peste Suína Clássica/diagnóstico , Ensaio de Imunoadsorção Enzimática/veterinária , Escherichia coli , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Suínos
11.
Zhonghua Nei Ke Za Zhi ; 61(9): 1044-1048, 2022 Sep 01.
Artigo em Chinês | MEDLINE | ID: mdl-36008298

RESUMO

To investigate the distribution and clinical significance of nuclear dense fine speckled (DFS) pattern in various diseases. A total of 95 289 patients who received DFS tests at Peking Union Medical College Hospital from January 2019 to December 2020 were included in this study. The results of indirect immunofluorescence assay (IIF) for detection of antinuclear antibody (ANA) were evaluated. The positive rates of ANA and DFS were 39.60% (37 733/95 289) and 1.19% (1 139/95 289) respectively. The positive rate of DFS in ANA-positive patients was 3.02% (1 139/37 733). DFS and ANA positivity were significantly different among different age groups rather than gender. The positivity rate of DFS reached the peak (55.57%, 633/1 139) in young patients between 21-40 years, while positive ANA with negative DFS was mainly observed in patients between 41-60 years (37.26%, 13 636/36 594). Additionally, single ANA-positivity were mainly detected in rheumatology department (59.23%, 18 402/31 066), whereas positive DFS was more common in obstetrics and gynecology department (3.08%, 49/1 593). There were 82.88% (944/1 139) patients with positive DFS diagnosed with non-autoimmune disease (non-AID), and 19.49%(222/1 139) with dermatosis. Positive DFS with higher titer (≥1∶320) was detected more frequently in autoimmune disease (AID) patients (5.13%, 10/195) than in non-AID patients (1.69%, 16/944) (P<0.05). The DFS pattern is rare in ANA positive patients, which is mainly observed in women between 21-49 years. High titer of DFS is prevalent in AID patients, but positive DFS is detected more in non-AID patients, especially those with dermatosis.


Assuntos
Doenças Autoimunes , Dermatopatias , Proteínas Adaptadoras de Transdução de Sinal , Anticorpos Antinucleares , Doenças Autoimunes/diagnóstico , Feminino , Técnica Indireta de Fluorescência para Anticorpo/métodos , Humanos , Fatores de Transcrição
13.
Am J Pathol ; 192(9): 1295-1304, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35750258

RESUMO

The detection of serum Epstein-Barr virus antibodies by immunofluorescence assay (IFA) is considered the gold standard screening test for nasopharyngeal cancer (NPC) in high-risk populations. Given the high survival rate after early detection in asymptomatic patients, compared to the poor prognosis in patients with late-stage NPC, screening using IFA has tremendous potential for saving lives in the general population. However, IFA requires visual interpretation of cellular staining patterns by trained pathology staff, making it labor intensive and hence nonscalable. In this study, an automated fuzzy inference (FI) system achieved high agreement with a human IFA expert in identifying cellular patterns associated with NPC (κ = 0.82). The integration of a deep learning module into FI further improved the performance of FI (κ = 0.90) and reduced the number of uncertain cases that required manual evaluation. The performance of the resulting hybrid model, termed deep learning FI (DeLFI), was then evaluated with a separate testing set of clinical samples. In this clinical validation, DeLFI outperformed human evaluation on the area under the curve (0.926 versus 0.821) and closely matched human performance on Youden J index (0.81 versus 0.80). Data from this study indicate that the combination of deep learning with FI in DeLFI has the potential to improve the scalability and accuracy of NPC detection.


Assuntos
Aprendizado Profundo , Infecções por Vírus Epstein-Barr , Neoplasias Nasofaríngeas , Técnica Indireta de Fluorescência para Anticorpo , Herpesvirus Humano 4 , Humanos , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/diagnóstico
14.
J Clin Lab Anal ; 36(7): e24554, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35708068

RESUMO

BACKGROUND: Antinuclear antibodies (ANAs) are invaluable biomarkers for the diagnosis of autoimmune diseases (AIDs). This study aims to compare the performances of line immunoassay (LIA), multiplex bead-based flow fluorescent immunoassay (MBFFI), and magnetic bar code immunofluorescence assay (MBC-IF) to detect ANA-Profile-15S. METHODS: In total, 184 samples from AID patients and 50 healthy controls (HCs) were collected. Fifteen ANAs (anti-dsDNA, nucleosome, histone, Sm, PCNA, ribosomal-P, SS-A/Ro52, SS-A/Ro60, SS-B/La, centromere B [CENP-B], Scl-70, U1-snRNP, AMA-M2, Jo-1, and Pm/Scl) were subjected to parallel detection by the LIA, MBFFI, and MBC-IF. The consistency between assays was analyzed. The discrepant results were further examined by chemiluminescent immunoassay (CLIA). RESULTS: Anti-SS-A/Ro52 and SS-A/Ro60 autoantibodies were the most common autoantibodies in ANA positive-profiles, and were detected with equal efficiency by the LIA, MBFFI, and MBC-IF (p = 0.101 and p = 0.732, respectively). The three assays showed excellent agreement (consistency range: 66.5%-97.5%), and total consistency was 85.8%. The MBFFI and MBC-IF assays were in good agreement in terms of ANA-Profile-15S determination; the kappa coefficient ranged from 0.59 to 0.95, except for the PCNA and PM-Scl. Of the 262 re-assessed divergent results, 124 (47.33%) were positive on CLIA; the various autoantibodies exhibited variable patterns. More importantly, the ANA-Profile-15S results of the MBFFI and MBC-IF accurately identified patients with AID; the area under the curves ranged from 0.642 to 0.919. CONCLUSIONS: The novel MBFFI and MBC-IF assay performed well in detecting ANA-Profile-15S. The application of MBFFI and MBC-IF play important roles in laboratory diagnosis of AIDs.


Assuntos
Síndrome de Imunodeficiência Adquirida , Doenças Autoimunes , Anticorpos Antinucleares , Autoanticorpos , Doenças Autoimunes/diagnóstico , Técnica Indireta de Fluorescência para Anticorpo/métodos , Humanos , Antígeno Nuclear de Célula em Proliferação
15.
J Clin Microbiol ; 60(7): e0023622, 2022 07 20.
Artigo em Inglês | MEDLINE | ID: mdl-35652310

RESUMO

Kangaroos are considered to be an important reservoir of Q fever in Australia, although there is limited knowledge on the true prevalence and distribution of coxiellosis in Australian macropod populations. Serological tests serve as useful surveillance tools, but formal test validation is needed to be able to estimate true seroprevalence rates, and few tests have been validated to screen wildlife species for Q fever. In this study, we modified and optimized a phase-specific indirect immunofluorescence assay (IFA) for the detection of IgG antibodies against Coxiella burnetii in macropod sera. The assay was validated against the commercially available ID Screen Q fever indirect multispecies enzyme-linked immunosorbent assay (ELISA) kit (IDVet, Grabels, France) to estimate the diagnostic sensitivity and specificity of each assay, using Bayesian latent class analysis. A direct comparison of the two tests was performed by testing 303 serum samples from 10 macropod populations from the east coast of Australia and New Zealand. The analysis indicated that the IFA had relatively high diagnostic sensitivity (97.6% [95% credible interval [CrI], 88.0 to 99.9]) and diagnostic specificity (98.5% [95% CrI, 94.4 to 99.9]). In comparison, the ELISA had relatively poor diagnostic sensitivity (42.1% [95% CrI, 33.7 to 50.8]) and similar diagnostic specificity (99.2% [95% CrI, 96.4 to 100]) using the cutoff values recommended by the manufacturer. The estimated true seroprevalence of C. burnetii exposure in the macropod populations included in this study ranged from 0% in New Zealand and Victoria, Australia, up to 94.2% in one population from New South Wales, Australia.


Assuntos
Coxiella burnetii , Febre Q , Anticorpos Antibacterianos , Teorema de Bayes , Ensaio de Imunoadsorção Enzimática , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Febre Q/diagnóstico , Febre Q/epidemiologia , Febre Q/veterinária , Estudos Soroepidemiológicos , Vitória
16.
Vet Dermatol ; 33(5): 414-425, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35670648

RESUMO

BACKGROUND: Canine trunk-dominant pemphigus foliaceus (PF) is mentioned rarely in the literature. HYPOTHESIS/OBJECTIVES: The goal of this study was to provide clinical description of trunk-dominant PF and to demonstrate the prevalence of serum antikeratinocyte, anti-desmocollin-1 (DSC1) and anti-desmoglein-1 (DSG1) antibodies, and determine their diagnostic value in this particular PF phenotype. MATERIALS AND METHODS: Clinically relevant information was collected from 31, 25 and 34 dogs with trunk-dominant and facial PF and superficial pyoderma (SP), respectively. Sera from these dogs were tested for antikeratinocyte, anti-DSC1 and anti-DSG1 antibodies using indirect immunofluorescence on canine tissues and DSC1- and DSG1-transfected cells. Sera from healthy dogs and dogs with clinically irrelevant diseases served as controls. RESULTS: Footpad involvement and grouped/polycyclic lesion organisation were identified as features of both PF phenotypes, and not of SP. Antikeratinocyte immunoglobulin (Ig)G was not specific for canine PF. By contrast, antigen-specific IgG was detected only in PF sera; anti-DSC1 IgG in 100% and 58% of dogs with facial and trunk-dominant PF, respectively, and anti-DSG1 IgG in 7% of dogs with trunk-dominant PF only. CONCLUSIONS: Trunk-dominant PF shares DSC1 as a major autoantigen with facial PF. The ability to detect anti-DSC1 IgG is lower in trunk-dominant PF, yet despite the lower sensitivity, the positive predictive value and accuracy of this particular anti-DSC1 IgG test are high. A negative test result, however, cannot exclude the diagnosis, and characteristic clinical features such as footpad involvement and/or grouped/polycyclic lesions must be considered when distinguishing trunk-dominant PF from its most relevant differential diagnosis: SP.


Contexte - Le pemphigus foliacé (PF) dominant le tronc chez le chien est rarement mentionné dans la littérature. Hypothèse/Objectifs - Le but de cette étude était de fournir une description clinique du PF dominant le tronc et de démontrer la prévalence des anticorps sériques anti-kératinocytes, anti-desmocolline-1 (DSC1) et anti-desmogléine-1 (DSG1), et de déterminer leur valeur diagnostique dans ce phénotype particulier de PF. Matériels et méthodes - Des informations cliniquement pertinentes ont été recueillies auprès de 31, 25 et 34 chiens atteints respectivement de PF à dominante tronculaire et faciale et de pyodermite superficielle (SP). Les sera de ces chiens ont été testés pour les anticorps anti-kératinocytes, anti-DSC1 et anti-DSG1 en utilisant l'immunofluorescence indirecte sur des tissus canins et des cellules transfectées avec DSC1 et DSG1. Des sera de chiens sains et de chiens atteints de maladies cliniquement non pertinentes ont servi de témoins. Résultats - L'implication du coussinet plantaire et l'organisation des lésions groupées / polycycliques ont été identifiées comme des caractéristiques des deux phénotypes PF, et non de SP. L'immunoglobuline antikératinocytaire (Ig)G n'était pas spécifique du PF canin. En revanche, l'IgG spécifique de l'antigène n'a été détectée que dans les sera de PF ; IgG anti-DSC1 chez 100 % et 58 % des chiens atteints de PF faciale et tronc-dominante, respectivement, et anti-DSG1 IgG chez 7 % des chiens avec PF tronc-dominant uniquement. Conclusions - Le PF à dominante tronculaire partage DSC1 comme auto-antigène majeur avec le PF facial. La capacité à détecter les IgG anti-DSC1 est plus faible chez les PF à dominante tronculaire, mais malgré la sensibilité plus faible, la valeur prédictive positive et la précision de ce test IgG anti-DSC1 particulier sont élevées. Cependant, un résultat de test négatif ne peut pas exclure le diagnostic, et les caractéristiques cliniques caractéristiques telles que l'atteinte du coussinet plantaire et/ou les lésions groupées/polycycliques doivent être prises en compte lors de la distinction entre la PF à dominante tronculaire et son diagnostic différentiel le plus pertinent : la SP.


Introducción- el pénfigo foliáceo (PF) de distribución truncal predominante se describe raramente n la literatura. Hipótesis/Objetivos- el objetivo de este estudio fue proporcionar una descripción clínica del PF truncal y demostrar la prevalencia de anticuerpos séricos antiqueratinocitos, antidesmocolina-1 (DSC1) y antidesmogleína-1 (DSG1), y determinar su valor diagnóstico en este fenotipo PF particular. Materiales y métodos- se recopiló información clínicamente relevante de 31, 25 y 34 perros con PF dominante truncal, PF dominante facial y pioderma superficial (PS), respectivamente. Los sueros de estos perros se analizaron en busca de anticuerpos antiqueratinocitos, anti-DSC1 y anti-DSG1 mediante inmunofluorescencia indirecta en tejidos caninos y células transfectadas con DSC1 y DSG1. Sueros de perros sanos y perros con enfermedades clínicamente irrelevantes sirvieron como controles. Resultados- la afectación de la almohadilla plantar y la organización de lesiones agrupadas/policíclicas se identificaron como características de ambos fenotipos de PF y no de SP. La inmunoglobulina (Ig)G antiqueratinocitos no fue específica para la PF canina. Por el contrario, la IgG específica de antígeno se detectó solo en sueros PF; IgG anti-DSC1 en el 100 % y el 58 % de los perros con PF predominante en la cara y el tronco, respectivamente, y IgG anti-DSG1 en el 7 % de los perros con PF predominante en el tronco solamente. Conclusiones- el PF dominante truncal comparte DSC1 como un autoantígeno importante con el PF facial. La capacidad para detectar IgG anti-DSC1 es menor en la PF dominante truncal; sin embargo, a pesar de la menor sensibilidad, el valor predictivo positivo y la precisión de esta prueba de IgG anti-DSC1 en particular son altos. Sin embargo, un resultado negativo de la prueba no puede excluir el diagnóstico, y deben tenerse en cuenta las características clínicas, como la afectación de las almohadillas plantares y/o las lesiones agrupadas/policíclicas, al distinguir la PF dominante truncal de su diagnóstico diferencial más relevante: SP.


Contexto - O pênfigo foliáceo (PF) canino predominante no tronco é raramente relatado na literatura. Hipótese/Objetivos - O objetivo deste estudo foi apresentar a descrição clínica do PF predominante no tronco e demonstrar a prevalência de anticorpos anti-queratinócitos, anti-desmocolina-1 (DSC1) e anti-desmogleína-1 (DSG1), e determinar o seu potencial diagnóstico neste tipo particular de PF. Materiais e métodos - Informações clinicamente relevantes foram coletadas de 31, 25 e 34 cães com PF predominante no tronco, PF facial e piodermite superficial (PS), respectivamente. Os soros destes cães foram testados para anticorpos anti-queratinócitos, anti-DSC1 e anti-DSG1 utilizando imunofluorescência indireta em tecidos caninos e em células DSC1 e DSG1 transfectadas. Os soros de cães saudáveis e cães com doenças clinicamente irrelevantes serviram de controle. Resultados - O acometimento dos coxins e a organização agrupada/policíclica das lesões foram identificadas como características de ambos os fenótipos de PF, não de PS. A imunoglobulina (Ig)G anti-queratinócitos não foi específica para PF. Em contraste, IgG antígeno-específica foi detectada apenas no soro de PF; IgG anti-DSC1 em 100% e 58% dos cães com PF facial e predominante no tronco, respectivamente, e IgG anti-DSG1 em 7% dos cães somente acometidos pelo PF predominante no tronco. Conclusões - O PF predominante no tronco compartilha DSC1 como um autoantígeno principal com PF facial. A capacidade de detectar IgG anti-DSC1 é menor no PF predominante no tronco, mas apesar da sensibilidade mais baixa, o valor preditivo positivo e a precisão do teste IgG anti-DSC1 específico são altos. Um resultado de teste negativo, no entanto, não pode excluir o diagnóstico, e características clínicas típicas, como envolvimento do coxim plantar e/ou lesões agrupadas/policíclicas, devem ser consideradas ao distinguir PF predominante no tronco de seu diagnóstico diferencial mais relevante: PS.


Assuntos
Doenças do Cão , Pênfigo , Animais , Autoanticorpos , Desmogleína 1 , Cães , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Imunoglobulina G , Pênfigo/diagnóstico , Pênfigo/veterinária
17.
Int J Rheum Dis ; 25(9): 991-1002, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35726725

RESUMO

OBJECTIVE: The primary aim of this study was to conduct a meta-analysis to compare the diagnostic accuracy of automated indirect immunofluorescence (automated-IIF) and fully automated solid-phase immunoassays (solid-phase assays), compared with gold standard conventional manual indirect immunofluorescence (manual-IIF) for antinuclear antibody (ANA) detection. METHODS: Indirect meta-comparison was performed using prospective studies reporting comparative data between automated-IIF and fully automated solid-phase assays individually to conventional manual-IIF. Diagnostic tests regarding different automated solid-phase assays and automated-IIF for ANA detection were retrieved from the Cochrane Library, PubMed, Embase, Web of Science, Chinese Biological Medicine Database (CBM), China National Knowledge Infrastructure (CNKI), and WANFANG electronic databases from their inception to January 2022. Assessment of the quality of the studies was undertaken using a second version of the Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) tool. The investigated diagnostic indices including pooled sensitivity, pooled specificity, pooled positive likelihood ratio (PLR), pooled negative likelihood ratio (NLR), pooled diagnostic odds ratio (DOR), area under the summary receiver operating characteristic (AUC) of automated-IIF and solid-phase assays, respectively. Relative diagnostic odds ratio (RDOR) was calculated to indirectly compare the diagnostic accuracy of automated-IIF and solid-phase assays. To visualize results, we provide forest plots showing the RDOR with 95% confidence intervals (CI) of the 2 methods against the "gold standard" manual-IIF by R software. Deeks' funnel was used to investigate the publication bias. RESULTS: A total of 16 studies involving 6111 subjects were included in the analysis. The pooled sensitivity, pooled specificity, pooled PLR, pooled NLR, pooled DOR and the AUC were 0.85 (95% CI: 0.84-0.86), 0.82 (95% CI 0.81-0.84), 14.22 (95% CI 8.55-23.65), 0.06 (95% CI 0.03-0.12), 287.0 (95% CI 124.30-662.68) and 0.983 for automated-IIF respectively, and as for solid-phase assays those were 0.73 (95% CI 0.70-0.75), 0.87 (95% CI 0.85-0.89), 5.66 (95% CI 3.33-9.62), 0.30 (95% CI 0.20-0.47), 19.14 (95% CI 8.00-45.79) and 0.894. The results of indirect comparison indicated that automated-IIF had statistically significant higher accuracy for the detection of ANA. CONCLUSION: This meta-analysis and indirect comparison suggest that automated-IIF should be recommended as an alternative assay for ANA screening under the condition of increased demand for ANA testing in clinical immunology laboratories.


Assuntos
Anticorpos Antinucleares , Testes Diagnósticos de Rotina , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Imunoensaio , Estudos Prospectivos , Sensibilidade e Especificidade
18.
J Infect Dev Ctries ; 16(4): 712-716, 2022 04 30.
Artigo em Inglês | MEDLINE | ID: mdl-35544635

RESUMO

The hepatosplenic (HS) form of cat scratch disease (CSD) is rarely seen; however, management of the treatment is challenging for clinicians. Monotherapy or combination regimens may be preferred based on severity of cases. Along with that, there are uncertainties as to the combination and duration of antibiotics effective against the microorganisms. In this report, a 12-year-old girl diagnosed with HS-CSD and unresponsive to primary treatment with macrolide group antibiotic was presented. The patient had liver findings compatible with CSD, confirmed radiologically and pathologically, and Bartonella henselae indirect immunofluorescence assay IgG was positive at 1/2048 titre. A combination therapy for six months with doxycycline and rifampicin was initiated, and the patient was successfully treated. The preference for monotherapy or combination regimen in HS-CSD is predominantly determined by the clinician according to the severity of the patient's clinical findings. The effectivity of antimicrobial regimen in HS-CSD requires further investigation.


Assuntos
Bartonella henselae , Doença da Arranhadura de Gato , Antibacterianos/uso terapêutico , Anticorpos Antibacterianos , Doença da Arranhadura de Gato/diagnóstico , Doença da Arranhadura de Gato/tratamento farmacológico , Criança , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Imunoglobulina G
19.
Exp Parasitol ; 238: 108284, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35609626

RESUMO

Toxocariasis is caused by infection with the nematode species Toxocara canis and Toxocara cati. Serological methods using eggs, larvae and adult worms of Toxocara spp. as antigen have been used for the diagnosis of human toxocariasis. The current study aimed to evaluate indirect immunofluorescence assay (IFA) using embryonated eggs of Toxocara for diagnosis of human toxocariasis. A total of 58 sera including twenty sera from patients with toxocariasis, 20 from healthy persons and 18 from patients with other parasitic infections were collected and used for the study. The embryonated eggs of Toxocara were prepared as antigen. Indirect immunofluorescence assay was performed using the frozen section of uterus containing embryonated T. canis eggs and unembryonated T. cati eggs. All serum samples had a positive reaction using IFA. The eggs of Toxocara as antigen exposed to the serum samples of toxocariasis, other parasitic infections and healthy persons, followed by IFA gave a bright greenish-yellow fluorescence. A number of samples such as eggs of Toxocara, Toxascaris, Trichuris and strongyloides larvae, and adult worm of Ancylostoma exhibited the bright greenish-yellow autofluorescence under fluorescent microscope. IFA using cryocut of embryonated eggs of Toxocara cannot be used for the diagnosis of human toxocariasis due to the existence of autofluorescence of the unembryonated and embryonated eggs, the second stage larva and adult worms of Toxocara spp.


Assuntos
Toxocara canis , Toxocaríase , Animais , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Larva , Toxocara , Toxocaríase/parasitologia
20.
Sci Rep ; 12(1): 6020, 2022 04 11.
Artigo em Inglês | MEDLINE | ID: mdl-35411013

RESUMO

Sjögren's syndrome (SS) is a common chronic inflammatory autoimmune disease that affects about 0.33-0.77% population in China. The positive for antinuclear antibodies (ANA) is one of the key features of SS, which shows a nuclear fine speckled (AC-4) pattern in an indirect immunofluorescent antibody test (IIFT). About 70% of ANA-positive SS patients have detectable anti-SS-A and/or SS-B antibodies, which indicates that other autoantibodies may present in SS patients. The anti-HMGB1 antibodies in 93 SS patients and 96 healthy controls were investigated with in-house developed ELISA and immunoblotting, and the locations of HMGB1 and fluorescent pattern of anti-HMGB1 antibody were investigated with IIFT. The contribution of anti-HMGB1 antibody in ANA-IF was evaluated with Cas9-induce HMGB1 knockout B16 cells. The anti-HMGB1 antibody level is higher in SS patients (9.96 ± 5.55 RU/ml) than in healthy controls (4.9 ± 1.4 RU/ml). With ROC curve analysis, when taking 8 RU/ml as the cutoff value, the sensitivity, specificity, and the area under the curve were 64.5%, 96.9%, and 0.83, respectively. A total of 18 patients (20.7%) with nuclear fine speckled (AC-4) pattern in ANA-IF test were anti-HMGB1 antibody positive only. With commercial antibody, anti-HMGB1 antibody showed the same nuclear fine speckled (AC-4) pattern. The serum from ANA-IF (+), SS-A (-), and SS-B (-) SS patients showed nuclear fine speckled (AC-4) pattern in wildtype B16 cells, but no fluorescence in HMGB1 knockout B16 cells. Anti-HMGB1 antibody may be one of the characteristic autoantibodies of SS in addition to anti-SS-A and SS-B. The detection of anti-HMGB1 antibody can provide more laboratory evidence for clinical diagnosis of SS.


Assuntos
Proteína HMGB1 , Síndrome de Sjogren , Anticorpos Antinucleares , Autoanticorpos , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Síndrome de Sjogren/diagnóstico
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