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Abstract Background: The VI Brazilian Consensus on Autoantibodies against HEp-2 cells for determination of autoantibodies against cellular constituents on HEp-2 cells was held on September, 2019, in Fortaleza (CE, Brazil). The guidelines in this edition were formulated by the group of Brazilian experts discussing the classification of complex patterns, the classification of the nuclear discrete dots (few and multiple), the identification of the discrete fine speckled pattern (AC-4a) and improvements on the ANA report. Mainbody: Sixteen Brazilian researchers and experts from universities and clinical laboratories representing the various geographical regions of Brazil participated in the meeting. Four main topics were discussed: (1) How to classify patterns with fluorescence in more than one cell compartment considering three relevant categoris: composite patterns, mixed patterns and multiple patterns; (2) The splitting of the discrete nuclear dots pattern into the multiple discrete nuclear dots (AC-6) and few discrete nuclear dots (AC-7) patterns, respectively; (3) Inclusion of a novel nuclear pattern characterized by discrete fine speckled pattern highly associated with antibodies to SS-A/Ro60, classified as AC-4a. In addition, adjustments on the Brazilian Consensus nomenclature were implemented aiming to harmonize the designation of some patterns with the International Consensus on ANA Patterns (ICAP). Furthermore, the designations of the PCNA-like pattern (AC-13), CENP-F-like pattern (AC-14) and Topo I-like pattern (AC-29) were adjusted in accordance to ICAP. Finally, there was a recommendation for adjustment in the test report in order to address the status of nuclear envelope staining. For all topics, the aim was to establish specific guidelines for laboratories and clinicians. All recommendations were based on consensus among participants. All recommendations from the V Consensus were maintained and there was relevant progress in the BCA/HEp-2 guidelines and further harmonization with ICAP. Conclusion: The VI BCA/HEp-2 edition was successful in establishing important recommendations regarding the classification of complex patterns, in supporting the identification of a novel pattern within the AC-4 group and in the harmonization process with the ICAP terminology.

https://doi.org/10.1186/s42358-022-00266-z

Investigation of nerve fibers in the skin by biopsy: technical aspects, indications, and contribution to diagnosis of small-fiber neuropathy
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Raicher, Irina

; Ravagnani, Luís Henrique Casartelli

; Correa, Silene Gomes

; Dobo, Cristine

; Mangueira, Cristóvão Luis Pitangueira

; Macarenco, Ricardo Silvestre e Silva

einstein (São Paulo)

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2022, Volume 20 elocation eMD8044

Abstract: En | Text: En | PDF: En

ABSTRACT Skin biopsy with investigation of small-diameter nerve fibers in human epidermis and dermis has been proven to be a useful method for confirming small-fiber neuropathy. In medical practice, small-fiber neuropathy is increasingly recognized as a leading cause of neuropathic pain. It is a prevalent complaint in medical offices, brought by patients often as a “painful burning sensation”. The prevalence of neuropathic pain is high in small-fiber neuropathies of different etiologies, especially in the elderly; 7% of population in this age group present peripheral neuropathy. Pain and paresthesia are symptoms which might cause disability and impair quality of life of patients. The early detection of small-fiber neuropathy can contribute to reducing unhealthy lifestyles, associated to higher incidence of the disease.

https://doi.org/10.31744/einstein_journal/2022md8044

Performance evaluation of an indirect immunofluorescence kit for the serological diagnosis of dengue
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Arai, Karina Emy

; Bo, Carolina Rodrigues Dal

; Silva, Ana Paula Marques Aguirra da

; Rodrigues, Silvia Sanches

; Mangueira, Cristóvão Luis Pitangueira

einstein (São Paulo)

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2020, Volume 18 elocation eAO5078

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

RESUMO Objetivo: Avaliar o desempenho da imunofluorescência indireta no diagnóstico sorológico de dengue em uma população com alta prevalência de arboviroses. Métodos: Duzentas amostras de soro de pacientes com suspeita clínica de dengue foram testadas por ensaio imunoenzimático e imunofluorescência indireta mosaico BIOCHIP®. Foram calculados especificidade, sensibilidade e coeficiente Kappa. Nas amostras discordantes, realizou-se reação em cadeia da polimerase como método confirmatório. Resultados: Das 200 amostras, 20% foram positivas e 80% negativas para IgM antivírus da dengue no ensaio imunoenzimático. Das 40 positivas, 25% foram negativas na imunofluorescência indireta. Destas dez negativas, apenas 20% eram também negativas na reação em cadeia da polimerase. Das 160 negativas no ensaio imunoenzimático, 5% foram positivas na imunofluorescência indireta. Por fim, dentre as nove discordantes, 33% tiveram vírus da dengue detectado na reação em cadeia da polimerase. O coeficiente Kappa foi 0,70 (0,57-0,82). Sensibilidade e especificidade por imunofluorescência indireta foram, respectivamente, 75% e 94%. Para IgG antivírus da dengue, de 200 amostras, 15,5% foram positivas e 84,5% negativas no ensaio imunoenzimático. Das 31 positivas, 12,9% foram negativas na imunofluorescência indireta. Destas quatro discordantes, 25% apresentaram vírus da dengue não detectado na reação em cadeia da polimerase. Das 169 negativas, 8% foram positivas na imunofluorescência indireta. Destas, 64% foram positivas também na reação em cadeia da polimerase. O coeficiente Kappa foi 0,695 (0,56-0,83). Sensibilidade e a especificidade por imunofluorescência indireta foram, respectivamente, 87,1% e 91,7%. Conclusão: Ensaio imunoenzimático seria suficiente para diagnóstico sorológico de infecção aguda, não justificando a incorporação da imunofluorescência indireta. Substituir ensaio imunoenzimático pela imunofluorescência indireta poderia comprometer a sensibilidade para IgM. Contudo, a imunofluorescência indireta auxilia diferenciar três arboviroses simultaneamente, sendo vantajoso em epidemias concomitantes.

ABSTRACT Objective: To evaluate the performance of indirect immunofluorescence for serological diagnosis of dengue virus in a population with high prevalence of arboviruses. Methods: Two-hundred serum samples from patients with clinical suspicion of dengue fever were tested by immunoenzymatic and indirect immunofluorescence assay BIOCHIP® mosaic. Specificity, sensitivity and Kappa coefficient were calculated. Discordant samples were tested by polymerase chain reaction for confirmation. Results: Of the 200 samples, 20% were positive and 80% negative for anti-dengue virus IgM antibodies in the immunoenzymatic test. Of the 40 positives, 25% were negative in indirect immunofluorescence. Of these ten discordant results, only 20% were also negative in the polymerase chain reaction (PCR). Of the 160 negatives in the immunoenzymatic test, 5% were positive in indirect immunofluorescence. Of these nine discordant results, 33% were positive in the PCR. The Kappa coefficient was 0.7 (0.572-0.829). Sensitivity and specificity of indirect immunofluorescence were respectively 75% and 94%. For anti-dengue virus IgG antibodies, of the 200 samples, 15.5% were positive and 84.5% were negative in the immunoenzymatic test. Of the 31 positives, 12.9% were negative in indirect immunofluorescence. Of these four discordant results, 25% were negative in the PCR. Of the 169 negatives, 8% were positive in indirect immunofluorescence. Of these 14 discordant results, 64% were also positive in the PCR. The Kappa coefficient was 0.695 (0.563-0.83). Sensitivity and specificity of indirect immunofluorescence were 87.1% and 91.7%, respectively. Conclusion: For diagnosis of acute infection, the immunoenzymatic test is enough, and the use of additional methods is not warranted. Replacing the immunoenzymatic test by indirect immunofluorescence would compromise the sensitivity for IgM. However, indirect immunofluorescence can distinguish three arboviruses simultaneously, an advantage during concomitant epidemics.

https://doi.org/10.31744/einstein_journal/2020ao5078 1744 downloads

Comparison between enzyme-linked immunosorbent assay and indirect immunofluorescence for detection of antineutrophil cytoplasmic antibodies
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Menezes, Julia Miranda

; Rossener, Raissa

; Silva, Ana Paula Marques Aguirra da

; Rodrigues, Silvia Sanches

; Mangueira, Cristóvão Luis Pitangueira

einstein (São Paulo)

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2020, Volume 18 elocation eAO5132

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

RESUMO Objetivo Avaliar o desempenho das metodologias de ensaio imunoenzimático e imunofluorescência indireta para a detecção de anticorpos anticitoplasma de neutrófilos em um contexto de laboratório clínico de rotina. Métodos Foram testadas 227 amostras pelas metodologias de imunofluorescência indireta e ensaio imunoenzimático com especificidades para anticorpos antiproteinase-3 e antimieloperoxidase. As proporções de amostras positivas foram comparadas por hipóteses de McNemar, e a concordância foi descrita pelo coeficiente Kappa de Cohen. Resultados A concordância dos testes foi 96,5%, e o coeficiente Kappa obtido foi 0,70 (IC95%: 0,50-0,90; p<0,001). Utilizando a imunofluorescência indireta como padrão-ouro, a sensibilidade do ensaio imunoenzimático foi de 0,62 e a especificidade, 0,99, com acurácia diagnóstica em 96% dos casos. Algumas amostras apresentaram resultados negativos por ensaio imunoenzimático e positivos por imunofluorescência. Isso ocorreu em amostras com vários padrões de fluorescência, mas particularmente nos casos com padrões atípicos. Duas amostras com positividade antiproteinase 3 foram consideradas negativas por imunofluorescência. Conclusão Os métodos de ensaio imunoenzimático tiveram alta especificidade, mas sensibilidade inferior. A realização da imunofluorescência indireta aumenta a sensibilidade diagnóstica, ao mesmo tempo que a pesquisa de antiproteinase 3 por ensaio imunoenzimático também pode agregar poder diagnóstico.

ABSTRACT Objective To evaluate the performance of enzyme-linked immunosorbent assay and indirect immunofluorescence methods for the detection of antineutrophil cytoplasmic antibodies in a routine clinical laboratory setting. Methods A total of 227 samples were tested by indirect immunofluorescence and enzyme-linked immunosorbent assay with antigen specificity for antiproteinase 3 and antimyeloperoxidase. The proportions of positive samples were compared by McNemar hypotheses and agreement was described by Cohen’s Kappa coefficient. Results The agreement of the tests was 96.5%, and the Kappa coefficient obtained was 0.70 (95%CI: 0.50-0.90; p<0.001). Considering indirect immunofluorescence as the gold standard, the sensitivity of the enzyme-linked immunosorbent assay was 0.62 and the specificity was 0.99, with diagnostic accuracy in 96% of cases. Some samples were negative in enzyme-linked immunosorbent assay and positive in indirect immunofluorescence. This situation occurred in all immunofluorescence patterns, but particularly in atypical patterns. Two samples with antiproteinase 3 positivity were considered negative in indirect immunofluorescence. Conclusion The enzyme-linked immunosorbent assay had high specificity but lower sensitivity. The performance of indirect immunofluorescence increases diagnostic sensitivity, while the search for antiproteinase 3 by enzyme-linked immunosorbent assay may also add diagnostic power.

https://doi.org/10.31744/einstein_journal/2020ao5132 1373 downloads

V Brazilian consensus guidelines for detection of anti-cell autoantibodies on hep-2 cells
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Cruvinel, Wilson de Melo

; Andrade, Luis Eduardo Coelho

; von Mühlen, Carlos Alberto

; Dellavance, Alessandra

; Ximenes, Antônio Carlos

; Bichara, Carlos David

; Bueno, Cleonice

; Mangueira, Cristóvão Luis Pitangueira

; Bonfá, Eloísa

; Brito, Fabiano de Almeida

; Flumian, Fernanda Bull

; Silva, Glaucielen Gomes da

; Rêgo, Jozelia

; Anjos, Lisiane Maria Ericoni dos

; Slhessarenko, Natasha

; Pasoto, Sandra Gofinet

; Neves, Suzane Pretti Figueiredo

; Valim, Valéria

; Santos, Wilton Silva dos

; Francescantonio, Paulo Luiz Carvalho

Advances in Rheumatology

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2019, Volume 59 elocation 28

Abstract: En | Text: En | PDF: En

Abstract Background: The V Brazilian Consensus for determination of autoantibodies against cellular constituents on HEp-2 cells, held in Brasilia (DF, Brazil) on August 27, 2016, discussed the harmonization between the Brazilian Consensus on ANA (BCA) guidelines and the International Consensus on ANA Patterns (ICAP) recommendations (www.anapatterns.org). Initial guidelines were formulated by the group of Brazilian experts with the purpose of guiding and enabling Brazilian clinical laboratories to adopt recommendations and to provide a common standard for national and international consensuses. Mainbody: Twenty Brazilian researchers and experts from universities and clinical laboratories representing the various geographical regions of the country participated in the meeting. Three main topics were discussed, namely the harmonization between the BCA guidelines and latest recommendations of the ICAP initiative, the adjustment of the terminology and report on HEp-2 patterns, and a reassessment of quality assurance parameters. For the three topics, our aim was to establish specific guidelines. All recommendations were based on consensus among participants. There was concrete progress in the adjustment of the BCA guidelines to match the ICAP guidelines. To a certain extent, this derives from the fact that ICAP recommendations were largely based on the algorithm and recommendations of the IV Brazilian ANA Consensus, as consistently recognized in the ICAP publications and presentations. However, although there is great overlap between the two Consensuses, there are some point divergences. These specific items were individually and extensively discussed, and it was acknowledged that in several points ICAP improved recommendations previously issued by the Brazilian ANA Consensus and these changes were readily implemented. Regarding some specific topics, the BCA panel of experts felt that the previously issued recommendations remained relevant and possibly will require further discussion with ICAP. The term anti-cell antibodies was adopted as the recommended designation, recognizing that the assay addresses antibodies against antigens in the nucleus and in other cell compartments. However, the acronym ANA HEp-2 was maintained due to historical and regulatory reasons. It was also signalized that the latest trend in ICAP is to adopt the term Indirect Immunofluorescent Assay on HEp-2 cell substrate (HEp-2 IIFA). In addition, the quality assurance strategies previously presented were ratified and emphasized. Conclusion: The V BCA edition was successful in establishing an overall harmonization with the ICAP recommendations for interpretation of the HEp-2 IIFA test, pinpointing the perspectives in filling the remaining gaps between both initiatives.

https://doi.org/10.1186/s42358-019-0069-5 1618 downloads

Glicemia de jejum de pacientes da rede pública de saúde na região sul de São Paulo: correlação com hemoglobina glicada e níveis lipídicos
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Franco, Luciana Ferreira

; Mafra, Ana Carolina Cintra Nunes

; Bracco, Mario Maia

; Franco, Laercio Joel

; Naves, Larissa Kozloff

; Ribeiro, Glória Maria Ferreira

; Mangueira, Cristóvão Luis Pitangueira

Revista Brasileira de Epidemiologia

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2019, Volume 22 elocation e190058

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

RESUMO: Introdução: A glicemia de jejum é um teste usado para o monitoramento do diabetes mellitus, bem como para seu rastreamento e diagnóstico. O objetivo do estudo foi analisar resultados de glicemia de jejum de pacientes da rede pública e sua correlação com hemoglobina glicada e lipídios. Métodos: Estudo transversal, com 77.581 pacientes, atendidos em 2014. Resultados: A maioria é do sexo feminino (65%), com idade entre 18 e 115 anos (53 ± 15,5 anos). A concordância entre glicemia de jejum e hemoglobina glicada foi moderada (Kappa = 0,416), entretanto foi substancial para níveis compatíveis com diabetes (Kappa = 0,689) e pobre para pré-diabetes (Kappa = 0,188). Glicemia de jejum ≥ 100 mg/dL foi encontrada em 41,1% dos pacientes e hemoglobina glicada ≥ 5,7% em 61,5%. As alterações lipídicas são mais frequentes nos indivíduos com alterações na glicemia. Dos 14.241 indivíduos com glicemia de jejum ≥ 126 mg/dL, a microalbuminúria foi pesquisada em apenas 883 (6,2%) indivíduos, com resultado alterado em 201 (22,8%). Conclusões: Nos indivíduos que realizaram mais de uma dosagem de glicemia de jejum, a maioria permaneceu com exames alterados, principalmente os que apresentavam valores compatíveis com o diagnóstico de diabetes, sugerindo que não conseguem um controle adequado. A baixa frequência de pesquisa de microalbuminúria em indivíduos com glicemia de jejum sugestiva de diabetes reflete a pequena preocupação com o rastreio de suas complicações crônicas. A elevada frequência de dislipidemia nesses indivíduos evidencia ser uma população de elevado risco cardiovascular.

ABSTRACT: Introduction: Fasting glucose is a test used for monitoring diabetes mellitus, as well as its screening and diagnosis. The objective of this study was to evaluate fasting glucose results and their correlation with glycated hemoglobin and lipids. Methods: Cross-sectional study, involving 77,581 patients, attended in 2014. Results: The majority of the patients are women (65%). The age of the patients ranged from 18 to 115 years (mean of 53 ± 15.5). The agreement between fasting glucose and glycated hemoglobin was moderate (kappa = 0.416); however, it was substantial for the levels used for the diagnosis of diabetes (kappa = 0.689) and poor for pre-diabetes (kappa = 0.188). Fasting glucose ≥ 100 mg/dL was observed in 41.1% of the patients and 61.5% present glycated hemoglobin ≥ 5.7%. Lipid abnormalities are likeliest in patients with elevated fasting glucose. From those 14,241 individuals that had fasting glucose ≥ 126 mg/dL, the microalbuminuria test was performed in only 883 (6.2%) patients, with abnormal results in 201 (22.8%). Conclusions: The high frequency of fasting glucose with abnormal results may reflect the high proportion of exams performed by individuals with diagnosis of diabetes, to evaluate their glycemic control. The low frequency of requests for microalbuminuria tests in those with probable diagnosis of diabetes reflects the little attention paid for the screening of chronic complications of diabetes. It calls attention the high frequency of dyslipidemia in those individuals, highlighting the fact that this is a population with high cardiovascular risk.

https://doi.org/10.1590/1980-549720190058

Glicemia de jejum de pacientes da rede pública de saúde na região sul de São Paulo: correlação com hemoglobina glicada e níveis lipídicos
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Franco, Luciana Ferreira

; Mafra, Ana Carolina Cintra Nunes

; Bracco, Mario Maia

; Franco, Laercio Joel

; Naves, Larissa Kozloff

; Ribeiro, Glória Maria Ferreira

; Mangueira, Cristóvão Luis Pitangueira

Revista Brasileira de Epidemiologia

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2019, Volume 22 elocation e190058

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

https://doi.org/10.1590/1980-549720190058 1755 downloads

The cerebrospinal fluid in multiple sclerosis: far beyond the bands
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Domingues, Renan Barros

; Fernandes, Gustavo Bruniera Peres

; Leite, Fernando Brunale Vilela de Moura

; Tilbery, Charles Peter

; Thomaz, Rodrigo Barbosa

; Silva, Gisele Sampaio

; Mangueira, Cristóvão Luis Pitangueira

; Soares, Carlos Augusto Senne

einstein (São Paulo)

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mar 2017, Volume 15 N. 1 Pages 100 - 104

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

RESUMO A análise do líquido cefalorraquidiano tem sido empregada para avaliação diagnóstica da esclerose múltipla e a exclusão dos diagnósticos diferenciais. Os achados clássicos refletem a natureza inflamatória da doença, incluindo discreta pleocitose, leve hiperproteinorraquia, aumento da síntese intratecal de imunoglobulina G e, mais tipicamente, a presença de bandas oligoclonais. Nos últimos anos, surgiram novos biomarcadores para esclerose múltipla, e esta busca por marcadores reflete a necessidade de melhor avaliar a atividade e a progressão da doença, bem como a eficácia terapêutica. Uma avaliação mais refinada da atividade da doença e da resposta aos medicamentos pode contribuir para melhores decisões terapêuticas, particularmente no que se refere à troca de medicação. Isto é muito importante nos dias de hoje, quando surgem novas opções medicamentosas. Neste artigo de revisão, avaliamos criticamente a literatura atual referente aos novos marcadores liquóricos na esclerose múltipla. A mensuração destes marcadores, como a quimiocina CXCL13, fetuína A e, principalmente, o neurofilamento de cadeia leve, demonstrou resultados promissores na avaliação da doença, provendo informações que, em conjunto com dados clínicos e de neuroimagem, podem contribuir para melhores decisões terapêuticas.

ABSTRACT The cerebrospinal fluid analysis has been employed for supporting multiple sclerosis diagnosis and ruling out the differential diagnoses. The most classical findings reflect the inflammatory nature of the disease, including mild pleocytosis, mild protein increase, intrathecal synthesis of immunoglobulin G, and, most typically, the presence of oligoclonal bands. In recent years, new biomarkers have emerged in the context of multiple sclerosis. The search for new biomarkers reflect the need of a better evaluation of disease activity, disease progression, and treatment efficiency. A more refined evaluation of disease and therapy status can contribute to better therapeutic choices, particularly in escalation of therapies. This is very relevant taking into account the availability of a greater number of drugs for multiple sclerosis treatment in recent years. In this review, we critically evaluate the current literature regarding the most important cerebrospinal fluid biomarkers in multiple sclerosis. The determination of biomarkers levels, such as chemokine ligand 13, fetuin A, and mainly light neurofilament has shown promising results in the evaluation of this disease, providing information that along with clinical and neuroimaging data may contribute to better therapeutic decisions.

https://doi.org/10.1590/s1679-45082017rw3706 11304 downloads

Personalized medicine and the clinical laboratory
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Pinho, João Renato Rebello

; Sitnik, Roberta

; Mangueira, Cristóvão Luis Pitangueira

einstein (São Paulo)

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sep 2014, Volume 12 N. 3 Pages 366 - 373

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Medicina personalizada é o uso de biomarcadores, em sua maioria marcadores moleculares, para a detecção de traços genéticos específicos, a fim de orientar diversas abordagens para a prevenção e o tratamento de diferentes doenças. A identificação de vários genes relacionados a doenças hereditárias, oncológicas e infecciosas permite a detecção de polimorfismos genéticos que estão envolvidos em diferentes evoluções clínicas dessas doenças, bem como com variações na resposta ao tratamento. Atualmente, já é possível detectar esses polimorfismos utilizando diversas metodologias: a detecção de polimorfismos de nucleotídeo único pela reação de polimerização em cadeia; a detecção de microarranjos de ácidos nucleicos; e o sequenciamento de ácidos nucleicos com sequenciadores de DNA automatizados usando métodos derivados de sequenciamento Sanger ou de nova geração. Os ensaios de medicina personalizada são dirigidos para detectar variações genéticas que alteram interações de fármacos com alvos ou vias metabólicas de fármacos (anabólicas e catabólicas), podendo ser utilizados para a seleção de formulações farmacêuticas e para detectar diferentes imunogenicidades da droga. As aplicações de medicina personalizada já foram descritas em várias áreas da Medicina e permitem que abordagens de tratamento específicas sejam aplicadas para cada paciente e para cada doença, de acordo com os resultados dos ensaios utilizados. A aplicação de um protocolo desse tipo exige uma relação intensa entre o laboratório e o corpo clínico. Para sua execução, é necessária uma equipe coordenada, composta por investigadores de pesquisa básica e médicos altamente especializados em suas áreas, apoiada por um time bastante especializado de analistas clínicos treinados em testes de biologia molecular.

Personalized medicine is the use of biomarkers, most of them molecular markers, for detection of specific genetic traits to guide various approaches for preventing and treating different conditions. The identification of several genes related to heredity, oncology and infectious diseases lead to the detection of genetic polymorphisms that are involved not only in different clinical progression of these diseases but also in variations in treatment response. Currently, it is possible to detect these polymorphisms using several methodologies: detection of single nucleotide polymorphisms using polymerase chain reaction methods; nucleic acid microarray detection; and nucleic acid sequencing with automatized DNA sequencers using Sanger-derived methods and new generation sequencing. Personalized medicine assays are directed towards detecting genetic variations that alter interactions of drugs with targets or the metabolic pathways of drugs (upstream and downstream) and can be utilized for the selection of drug formulations and detect different immunogenicities of the drug. Personalized medicine applications have already been described in different areas of Medicine and allow specific treatment approaches to be applied to each patient and pathology according to the results of these assays. The application of such a protocol demands an increasing interaction between the clinical laboratory and the clinical staff. For its implementation, a coordinated team composed of basic researchers and physicians highly specialized in their areas supported by a highly specialized team of clinical analysts particularly trained in molecular biology assays is necessary.

https://doi.org/10.1590/s1679-45082014rw2859 11291 downloads

10.

SeptiFast for diagnosis of sepsis in severely ill patients from a Brazilian hospital
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Sitnik, Roberta

; Marra, Alexandre Rodrigues

; Petroni, Roberta Cardoso

; Ramos, Ozires Pereira Santos

; Martino, Marinês Dalla Valle

; Pasternak, Jacyr

; Santos, Oscar Fernando Pavão dos

; Mangueira, Cristóvão Luis Pitangueira

; Pinho, João Renato Rebello

einstein (São Paulo)

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jun 2014, Volume 12 N. 2 Pages 191 - 196

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Objetivo Testar e validar um método molecular multiplex para detecção de infecções sanguíneas, além de comparar os resultados com os obtidos pela hemocultura convencional.Métodos Os testes de hemocultura e o LightCycler® SeptiFast foram realizados em 114 pacientes consecutivos com evidência clínica de sepse.Resultados Mais amostras positivas (23; 20,2%) foram detectadas pelo LightCycler® SeptiFast do que pela hemocultura (17; 14,9%), mostrando concordância de 86,8%. Os resultados discordantes foram de quatro pacientes positivos apenas para hemocultura, dez positivos apenas para LightCycler® SeptiFast e um com patógenos diferentes encontrados em cada método. Infecções por micro-organismos não reconhecidos pelo LightCycler® SeptiFast e detectados apelas pela hemocultura confirmam a necessidade da realização dos dois métodos. O tempo médio para os resultados da hemocultura foi de 5 dias para amostras negativas e de 3,5 dias para as positivas. Os resultados pelo LightCycler® SeptiFast foram obtidos em menos de 8 horas.Conclusão O LightCycler® SeptiFast mostrou ser um teste de grande potencial para ser realizado simultaneamente à hemocultura para diagnóstico de sepse em doentes graves, permitindo um diagnóstico mais rápido de infecções por bactérias e fungos e, dessa forma, auxiliando a redução do tempo de hospitalização e racionalização do uso de antibióticos. Eventualmente, o LightCycler® SeptiFast pode detectar inclusive infecções por micro-organismos dificilmente detectáveis via hemocultura, especialmente aquelas causadas por Candida nãoalbicans.

Objective To test and validate a multiplex real-time polymerase chain reaction method for bloodstream infections, as well as to compare the results with conventional blood culture.Methods A total of 114 consecutive patients with clinical evidence of sepsis were submitted to blood culture and LightCycler™ SeptiFast tests.Results More positive specimens (23; 20.2%) were detected using the LightCycler™ SeptiFast than the blood culture (17; 14.9%), with an agreement of 86.8%. Discordant results were seen in four patients positive only to blood culture, ten positive only to LightCycler™ SeptiFast and one to different pathogens found by each test. Infections with microorganisms detected only using blood culture reassured the need to perform both tests. The mean time to results for blood culture was 5 days for negative and 3.5 days for positive results. LightCycler™ SeptiFast results were achieved in less than 8 hours.Conclusion LightCycler™ SeptiFast showed a high potential as a test to be carried out concomitantly with blood culture for sepsis diagnosis in severely ill patients. This test allowed a faster diagnosis of bacterial and fungal infections that helped to reduce hospital stay and to control the use of antibiotics. LightCycler™ SeptiFast can also eventually detect microorganism and infections that are hardly detected by blood culture, especiallyCandidanon-albicans infections.

https://doi.org/10.1590/S1679-45082014AO2932 5016 downloads

11.

IV Consenso Brasileiro para pesquisa de autoanticorpos em células HEp-2
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Francescantonio, Paulo Luiz Carvalho

; Cruvinel, Wilson de Melo

; Dellavance, Alessandra

; Andrade, Luis Eduardo Coelho

; Taliberti, Ben Hur

; von Mühlen, Carlos Alberto

; Bichara, Carlos David Araújo

; Bueno, Cleonice

; Mangueira, Cristóvão Luis Pitangueira

; Carvalho, Darlene Gonçalves

; Bonfá, Eloísa S.D. de O.

; Brito, Fabiano de Almeida

; Araújo, Flávia Ikeda e

; Rêgo, Jozelia

; Pereira, Kaline Medeiros Costa

; Anjos, Lisiane Maria Enriconi dos

; Bissoli, Maria de Fatima

; Santiago, Mittermayer Barreto

; Maluf, Natalya Zaidan

; Alvarenga, Rossana Rassi

; Neves, Suzane Pretti Figueiredo

; Valim, Valeria

; Santos, Wilton Silva dos

Revista Brasileira de Reumatologia

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feb 2014, Volume 54 N. 1 Pages 44 - 50

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Objetivo: O IV Consenso Brasileiro para Pesquisa de Autoanticorpos em Células HEp-2 (FAN) realizado em Vitória (ES), no dia 18 de setembro de 2012, objetivou discutir estratégias e recomendações relacionadas ao procedimento técnico, à padronização e à interpretação dos resultados da pesquisa de autoanticorpos em células HEp-2. Métodos: Participaram do evento 23 pesquisadores e especialistas de Universidades e laboratórios brasileiros. Foram abordados diferentes tópicos, discutidos amplamente a fim de se estabelecer recomendações específicas. Resultados e conclusão: O IV Consenso integrou à árvore de decisão o padrão citoplasmático em Anéis e Bastões, o padrão nuclear pontilhado Quasi-homogêneo (QH) e o padrão misto CENP-F. Discutiu-se ainda a necessidade de atenção para a classificação do padrão misto relacionado à presença de anticorpos anti-DNA topoisomerase I (Scl70), compreendendo os componentes nuclear pontilhado fino, nucleolar homogêneo, NOR na placa metafásica e citoplasmático pontilhado fino. Foram sugeridas diretrizes para o controle de qualidade do teste, diluição de triagem e diluição de esgotamento, e foi emitido alerta quanto à necessidade de atenção em relação à heterogeneidade de substratos disponíveis no mercado e a utilização de metodologias automatizadas para detecção de autoanticorpos.

Objective: The Fourth Brazilian Consensus for Autoantibodies Screening in HEp-2 Cells (ANA) was held in Vitória, Espírito Santo, and aimed to discuss strategies and recommendations about the technique, standardization, interpretation and quality control of the indirect immunofluorescence reaction on HEp-2 cells. Methods: Twenty three ANA experts from university centers and private laboratories in different areas from Brazil discussed and agreed upon recommendations for the fourth edition of the Brazilian Consensus for Autoantibodies Screening in HEp-2 Cells. Results and conclusion: The 4th ANA Consensus included three novel patterns into the existing algorithm (cytoplasmic Rods and Rings, nuclear Quasi-homogeneous, and CENP-F). Emphasis was given to the need of attention in describing the peculiar mixed pattern elicited by anti-DNA topoisomerase I (Scl-70) autoantibodies, comprising nuclear fine specked, nucleolar homogeneous pattern, NOR staining in metaphase plates, and cytoplasmic fine speckled patterns. The group also emphasized the need for continuous quality control in indirect immunofluorescence assays, the establishment of screening dilutions, as well as conjugate titration. An alert was made regarding the heterogeneity of commercial kits in defining patterns and the use of solid phase methodologies to determine the presence of autoantibodies.

https://doi.org/10.1016/j.rbr.2013.10.001 28365 downloads

12.

Anticorpos anti-C1q, anticromatina/nucleossomo e anti-dsDNA em pacientes com lúpus eritematoso sistêmico juvenil
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Jesus, Adriana Almeida de

; Campos, Lucia Maria Arruda

; Liphaus, Bernadete Lourdes

; Carneiro-Sampaio, Magda

; Mangueira, Cristóvão Luis Pitangueira

; Rosseto, Eliane Aparecida

; Silva, Clovis Artur Almeida da

; Scheinberg, Morton

Revista Brasileira de Reumatologia

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dec 2012, Volume 52 N. 6 Pages 976 - 981

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

OBJETIVO: Avaliar a presença de anticorpos anti-C1q, anticromatina/nucleossomo e anti-DNA de duplo filamento (dsDNA) em pacientes com lúpus eritematoso sistêmico juvenil (LESJ) e controles. MÉTODOS: Foram analisados 67 pacientes com LESJ e 34 controles saudáveis para presença de anticorpos anti-C1q, anticromatina/nucleossomo e anti-dsDNA pelo método ELISA. Os níveis de C1q foram avaliados por imunodifusão radial. RESULTADOS: Na época, a média de idade era similar entre os pacientes com LESJ e os controles (14,6 ± 3,86 vs. 13,6 ± 2,93 anos; P = 0,14). Foram observadas frequências mais altas de anticorpos anti-C1q, anticromatina/nucleossomo e anti-dsDNA em pacientes com LESJ em relação aos controles (20% vs. 0%; P = 0,0037; 48% vs. 0%; P < 0,0001 e 69% vs. 3%; P < 0,0001, respectivamente). A mediana dos anticorpos anti-C1q, anticromatina/nucleossomo e anti-dsDNA também foi significativamente mais alta em pacientes com LESJ em relação aos controles [9,6 (5,5-127) vs. 7,5 (5-20) unidades, P = 0,0006; 18 (1,9-212) vs. 3,2 (1,7-17) unidades, P < 0,0001; e 111 UI/mL (6-741) vs. 14 (6-33) UI/mL, P < 0,0001, respectivamente]. A sensibilidade para os anticorpos anti-C1q, anticromatina/nucleossomo e anti-dsDNA foi: 21% (IC: 11-33), 49% (IC: 36-62) e 70% (IC: 57-81). A especificidade foi de 100% (IC: 88-100), 100% (88-100) e 97% (IC: 83-99), respectivamente. Foi observada uma correlação positiva entre os níveis de anti-dsDNA e tanto anticorpos anti-C1q (r = 0,51; IC: 0,29-0,68; P < 0,0001) como anticromatina/nucleossomo (r = 0,87; IC: 0,79-0,92; P < 0,0001). Foi observada uma correlação negativa entre os níveis de anti-C1q e C1q (r = -0,33; IC: -0,56-0,05; P = 0,018). A frequência de anti-dsDNA foi mais alta em pacientes com SLEDAI-2K > 1 (P = 0,0047), e não foram observadas diferenças nas frequências desses três autoanticorpos e nefrite (P > 0,05). CONCLUSÃO: Nosso estudo demonstrou elevada especificidade para diagnóstico de lúpus envolvendo os três autoanticorpos, especialmente anti-C1q e anticromatina/nucleossomo.

OBJECTIVES: To evaluate the presence of anti-C1q, anti-chromatin/nucleosome and anti-double stranded DNA (dsDNA) antibodies in juvenile systemic lupus erythematosus (JSLE) and controls. METHODS: Sixty-seven JSLE and 34 healthy controls were analyzed for the presence of anti-C1q, anti-chromatin/nucleosome, and anti-dsDNA antibodies by ELISA. C1q levels were evaluated by radial immunodiffusion. RESULTS: The mean current age was similar in JSLE patients and controls (14.6 ± 3.86 vs. 13.6 ± 2.93 years, P = 0.14). Higher frequencies of anti-C1q, anti-chromatin/nucleossome, and anti-dsDNA antibodies were observed in JSLE compared to controls (20% vs. 0%, P = 0.0037; 48% vs. 0%, P < 0.0001 and 69% vs. 3%, P < 0.0001, respectively). The median of anti-C1q, anti-chromatin/nucleossome, and antidsDNA antibodies were also significantly higher in JSLE patients than in controls [9.6 (5.5-127) vs. 7.5 (5-20) units, P = 0.0006; 18 (1.9-212) vs. 3.2 (1.7-17) units, P < 0.0001; and 111 IU/mL (6-741) vs. 14 (6-33) IU/mL; P < 0.0001, respectively]. The sensitivity for anti-C1q, anti-chromatin/nucleosome, and anti-dsDNA antibodies was 21% (CI: 11-33), 49% (CI: 36-62), and 70% (CI: 57-81). The specificity was 100% (CI: 88-100), 100% (88-100), and 97% (CI: 83-99), respectively. A positive correlation was found between anti-dsDNA levels and both anti-C1q (r = 0.51; CI: 0.29-0.68; P < 0.0001) and anti-chromatin/nucleosome antibodies (r = 0.87; CI: 0.79-0.92; P < 0.0001) levels. A negative correlation was observed between anti-C1q and C1q levels (r = -0.33; CI: -0.56-0.05; P = 0.018). The frequency of anti-dsDNA was higher in patients with SLEDAI-2K >1 (P = 0.0047) and no differences were observed in the frequencies of these three autoantibodies and nephritis (P > 0.05). CONCLUSION: Our study demonstrated an elevated specificity for lupus diagnosis involving the three autoantibodies, especially anti-C1q and anti-chromatin/nucleosome.

8810 downloads

13.

PCSK9 and its clinical importance with the new therapeutic targets against dyslipidemia
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Ferreira, Carlos Eduardo dos Santos

; Fonseca, Francisco Antônio Helfenstein

; Mangueira, Cristóvão Luis Pitangueira

einstein (São Paulo)

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dec 2012, Volume 10 N. 4 Pages 526 - 527

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Este é um progresso sensível; desde a descoberta das estatinas, não havia novas maneiras de diminuir, de maneira significativa, o colesterol e a fração LDL. Também está claro que essa redução, pelas estatinas, tem relação com futuras lesões cardiovasculares, sendo útil na profilaxia primária e secundária destas. Os autores apresentaram estudos sobre pesquisas para promover a queda do colesterol sanguíneo por meio de anticorpos que inibem a pró-proteína PCSK9, bem como agentes que atuam realizando a interferência no RNA. Duas vantagens se afiguram imediatamente: para pacientes que têm a miopatia relacionada às estatinas e por ser droga injetável a cada 15 dias, o que pode colaborar para maior adesão ao tratamento.

This is a remarkable progress; since the finding of statins, there was no new way of reducing, significantly, cholesterol and LDL fraction. It is also clear that this decrease, by statins, is related to future cardiovascular lesions, being useful in its primary and secondary prophylaxis. The authors presented studies on research to promote the falling of blood cholesterol by means of antibodies, which inhibit the pro-protein PCSK9, as well as agents that act performing the RNA interference. We had two advantages immediately: for patients with myopathy associated with statins, and the fact of being injected every 15 days, that may contribute to better treatment adherence.

11335 downloads

14.

III Consenso Brasileiro para Pesquisa de Autoanticorpos em Células HEp-2: perspectiva histórica, controle de qualidade e associações clínicas
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Francescantonio, Paulo Luiz Carvalho

; Andrade, Luis Eduardo Coelho

; Cruvinel, Wilson de Melo

; Araújo, Flávia Ikeda e

; Dellavance, Alessandra

; Gabriel Júnior, Alexandre

; Nuccitelli, Barbara

; Taliberti, Ben Hur

; von Mühlen, Carlos Alberto

; Bichara, Carlos David Araújo

; Santos, Cláudio Henrique Ramos dos

; Bueno, Cleonice

; Yano, Cristiane Martinez

; Mangueira, Cristóvão Luis Pitangueira

; Carvalho, Darlene Gonçalves

; Cardoso, Elizângela

; Bonfá, Eloísa

; Rassi, Gustavo Gabriel

; Mundim, Hugo Mendonça

; Bendet, Izidro

; Rego, Jozélia

; Vieira, Lisiane Maria Enriconi dos Anjos

; Barbosa, Maria Ordália Ferro

; Sugiyama, Mitiko

; Santiago, Mittermayer Barreto

; Slhessarenko, Natasha

; Silva, Nilzio Antônio da

; Jarach, Renata

; Suda, Roberto

; Levy, Roger Abramino

; Sampaio, Silvia Oliveira

; Neves, Suzane Pretti Figueiredo

; Santos, Wilton Silva dos

; Nóbrega, Yanna K. de M.

Jornal Brasileiro de Patologia e Medicina Laboratorial

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jun 2009, Volume 45 N. 3 Pages 185 - 199

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

OBJETIVO: O III Consenso Brasileiro para Pesquisa de Autoanticorpos em Células HEp-2 (FAN) objetivou discutir estratégias para controlar a qualidade do ensaio, promover a atualização das associações clínicas dos diversos padrões e avaliar as dificuldades de implantação do II Consenso ocorrido no ano de 2002. MÉTODOS: Nos dias 13 e 14 de abril de 2007 participaram do encontro em Goiânia pesquisadores e especialistas de diversos centros universitários e laboratórios clínicos de diferentes regiões do Brasil, com o propósito de discutir e aprovar as recomendações que visam a melhores padronização, interpretação e utilização do ensaio pelos clínicos. Foram convidados como ouvintes representantes comerciais de diferentes empresas produtoras de insumos para realização do teste de FAN. RESULTADOS E CONCLUSÃO: Dada a heterogeneidade de microscópios e reagentes disponíveis no mercado, o III Consenso enfatizou a necessidade do controle de qualidade em ensaios de imunofluorescência indireta. Foram também feitas algumas adequações na terminologia utilizada para classificar os diferentes padrões. Finalmente, foi realizada uma atualização das associações clínicas com finalidade de facilitar cada vez mais o melhor uso do ensaio pelos clínicos.

OBJECTIVE: The Third Brazilian Consensus for Autoantibodies Screening in HEp-2 Cells (ANA) had as purpose the evaluation of difficulties in the accomplishment of the 2nd Consensus recommendations that took place in the year of 2002, the discussion of strategies for quality control of the assay and the discussion of an update of the clinical associations of the several immunofluorescent patterns. METHODS: Several ANA experts from university centers and private laboratories in different areas in Brazil joined the workshop in Goiânia on 2007 April 13 and 14 with the purpose of discussing and approving the recommendations for standardization, interpretation and use of the test by physicians. Commercial representatives of different ANA slide brands were also invited as listeners to the workshop. RESULTS AND CONCLUSION: The 3rd ANA Consensus emphasized the need for quality control in indirect immunofluorescent assays since there is a considerable heterogeneity of available microscopes and reagents. It also promoted adaptations in the previously approved terminology used to classify the different patterns and finally updated the clinical associations of the several patterns with the purpose of providing guidance for interpretation of the assay by clinical pathologists and assistant physicians.

21533 downloads

15.

3º Consenso Brasileiro para pesquisa de autoanticorpos em células HEp-2 (FAN): recomendações para padronização do ensaio de pesquisa de autoanticorpos em células HEp-2, controle de qualidade e associações clínicas
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Dellavance, Alessandra

; Gabriel Júnior, Alexandre

; Nuccitelli, Barbara

; Taliberti, Ben Hur

; von Mühlen, Carlos Alberto

; Bichara, Carlos David Araújo

; Santos, Cláudio Henrique Ramos dos

; Bueno, Cleonice

; Yano, Cristiane Martinez

; Mangueira, Cristóvão Luis Pitangueira

; Carvalho, Darlene Gonçalves

; Cardoso, Elizângela

; Bonfá, Eloísa

; Araújo, Flávia Ikeda e

; Rassi, Gustavo Gabriel

; Mundim, Hugo Mendonça

; Bendet, Izidro

; Rego, Jozélia

; Vieira, Lisiane Maria Enriconi dos Anjos

; Andrade, Luis Eduardo Coelho

; Barbosa, Maria Ordália Ferro

; Sugiyama, Mitiko

; Santiago, Mittermayer Barreto

; Slhessarenko, Natasha

; Silva, Nilzio Antônio da

; Francescantonio, Paulo Luiz Carvalho

; Jarach, Renata

; Suda, Roberto

; Levy, Roger Abramino

; Sampaio, Silvia Oliveira

; Neves, Suzane Pretti Figueiredo

; Cruvinel, Wilson de Melo

; Santos, Wilton Silva dos

; Nóbrega, Yanna K. de M.

Revista Brasileira de Reumatologia

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apr 2009, Volume 49 N. 2 Pages 89 - 98

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

OBJETIVO:O 3º Consenso Brasileiro para pesquisa de autoanticorpos em Células HEp-2 (FAN) teve como propósito avaliar as dificuldades de implantação do 2º Consenso ocorrido no ano de 2002, discutir estratégias para controlar a qualidade do ensaio e promover a atualização das associações clínicas dos diversos padrões. MÉTODOS:Participaram do encontro em Goiânia nos dias 13 e 14 de abril de 2008 pesquisadores e especialistas de diversos centros universitários e laboratórios clínicos de diferentes regiões do Brasil, com o propósito de discutir e aprovar as recomendações que visam à melhor padronização, interpretação e utilização do ensaio pelos clínicos. Representantes comerciais de diferentes empresas produtoras de insumos para realização do teste de FAN foram convidados como ouvintes. RESULTADOS E CONCLUSÕES: O 3º Consenso enfatizou a necessidade do controle de qualidade em imunofluorescência dada a heterogeneidade de microscópios e reagentes disponíveis no mercado, promoveu adequações na terminologia utilizada para classificar os diferentes padrões e, finalmente, atualizou as associações clínicas com finalidade de facilitar cada vez mais o melhor uso do ensaio pelos clínicos.

OBJECTIVE: The Third Brazilian Consensus for autoantibodies Screening in HEp-2 cells had as purpose the evaluation of difficulties in the accomplishment of the 2nd Consensus recommendations that took place in the year of 2002, the discussion of strategies for quality control of the assay and the promotion of an update of the clinical associations of the several immunofluorescent patterns. METHODS:Several ANA experts from university centers and private laboratories in different areas in Brazil joined the workshop in Goiânia on 2008 April 13 and 14 with the purpose of discussing and approving the recommendations for standardization, interpretation and use of the test by physicians. Commercial representatives of different ANA slide brands were also invited as listeners to the workshop. RESULTS AND CONCLUSIONS: The 3rd Consensus emphasized the need for quality control in indirect immunofluorescent since there is a considerable heterogeneity of available microscopes and reagents. It also promoted adaptations in the previously approved terminology used to classify the different patterns and finally updated the clinical associations of the several patterns with the purpose of providing guidance for interpretation of the assay by clinical pathologists and assistant physicians.

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