title: Flow cytometric-based protocols for assessing anti-MT-2 IgG1 reactivity: High-dimensional data handling to define predictors for clinical follow-up of Human T-cell Leukemia virus type-1 infection.
abstract: The present work provides an innovative methodological approach to assess the anti-HTLV-1 IgG1 reactivity with practical application in clinical laboratory. Serum from non-infected healthy controls (NI) and HTLV-1-infected patients, categorized as asymptomatic (AS), putatively progressing to HTLV-1 associated myelopathy/tropical spastic paraparesis - HAM/TSP (pHAM) or with clinical diagnosis of HAM/TSP (HT) were assayed in two-parallel flow cytometry platforms, referred as: Fix and Fix&Perm protocols. Operating-characteristics analysis indicated that a single pair of attributes ("serum dilution/cut-off") for Fix and Fix&Perm protocols presented excellent performance for the diagnosis of HTLV-1 infection. Conversely, Fix and Fix&Perm protocols displayed weak/moderate overall performances when applied with prognosis purposes of HTLV-1 infection. A panoramic snapshot provided by the reactivity boards revealed clearly the higher sensitivity of Fix&Perm protocol for detecting seropositivity for HT, suggesting that stepwise combinatory criteria would improve the global performance of using a single pair of attributes. Three data mining strategies were tested, including endpoint titer analysis, heatmap assemblage and decision tree analysis. Bi-dimensional heatmap analysis demonstrated that, while the clustering profile of NI vs HTLV-1+revealed segregation in opposite poles, AS vs HT presented discrete segregation but still displaying an intertwined distribution pattern. The combination of methods for segregating AS from HT displayed a moderate but superior global accuracy (85.7%; LOOCV=71.4%). The comprehensive data analysis support that the combination of methods have improved the performance to the differential diagnosis of AS and HT, with direct association with laboratorial records, including serum cytokine levels and proviral load.
authors.affiliation: Grupo Integrado de Pesquisas em Biomarcadores - Centro de Pesquisas René Rachou - FIOCRUZ-Minas, Belo Horizonte, MG, Brazil. Electronic address: jordana.reis@cpqrr.fiocruz.br.
authors.forename: Jordana Grazziela
authors.initials: JG
authors.lastname: Coelho-Dos-Reis
authors.affiliation: Grupo Integrado de Pesquisas em Biomarcadores - Centro de Pesquisas René Rachou - FIOCRUZ-Minas, Belo Horizonte, MG, Brazil.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Peruhype-Magalhães
authors.affiliation: Grupo de Pesquisas Clínicas e Políticas Públicas em Doenças Infecciosas e Parasitárias - Centro de Pesquisas René Rachou - FIOCRUZ-Minas, Belo Horizonte, MG, Brazil.
authors.forename: Marcelo Antônio
authors.initials: MA
authors.lastname: Pascoal-Xavier
authors.affiliation: Laboratório de Bioinformática e Análise Molecular, Instituto de Genética e Bioquímica/Faculdade de Computação, Patos de Minas, MG, Brazil.
authors.forename: Matheus
authors.initials: M
authors.lastname: de Souza Gomes
authors.affiliation: Laboratório de Bioinformática e Análise Molecular, Instituto de Genética e Bioquímica/Faculdade de Computação, Patos de Minas, MG, Brazil.
authors.forename: Laurence Rodrigues
authors.initials: LR
authors.lastname: do Amaral
authors.affiliation: Grupo Integrado de Pesquisas em Biomarcadores - Centro de Pesquisas René Rachou - FIOCRUZ-Minas, Belo Horizonte, MG, Brazil.
authors.forename: Ludmila Melo
authors.initials: LM
authors.lastname: Cardoso
authors.affiliation: Grupo Integrado de Pesquisas em Biomarcadores - Centro de Pesquisas René Rachou - FIOCRUZ-Minas, Belo Horizonte, MG, Brazil.
authors.forename: Juan
authors.initials: J
authors.lastname: Jonathan-Gonçalves
authors.affiliation: Grupo Integrado de Pesquisas em Biomarcadores - Centro de Pesquisas René Rachou - FIOCRUZ-Minas, Belo Horizonte, MG, Brazil.
authors.forename: Ágata Lopes
authors.initials: ÁL
authors.lastname: Ribeiro
authors.affiliation: Fundação Centro de Hematologia e Hemoterapia de Minas Gerais - HEMOMINAS, Belo Horizonte, MG, Brazil; Programa de Pós-Graduação em Infectologia e Medicina Tropical - Faculdade de Medicina da Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
authors.forename: Ana Lúcia Borges
authors.initials: AL
authors.lastname: Starling
authors.affiliation: Hospital Sarah Kubitschek, Belo Horizonte, MG, Brazil.
authors.forename: João Gabriel
authors.initials: JG
authors.lastname: Ribas
authors.affiliation: Programa de Pós-Graduação em Infectologia e Medicina Tropical - Faculdade de Medicina da Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
authors.forename: Denise Utsch
authors.initials: DU
authors.lastname: Gonçalves
authors.affiliation: Fundação Centro de Hematologia e Hemoterapia de Minas Gerais - HEMOMINAS, Belo Horizonte, MG, Brazil.
authors.forename: Anna Bárbara
authors.initials: AB
authors.lastname: de Freitas Carneiro-Proietti
authors.affiliation: Grupo Integrado de Pesquisas em Biomarcadores - Centro de Pesquisas René Rachou - FIOCRUZ-Minas, Belo Horizonte, MG, Brazil.
authors.forename: Andréa
authors.initials: A
authors.lastname: Teixeira-Carvalho
authors.affiliation: Grupo Integrado de Pesquisas em Biomarcadores - Centro de Pesquisas René Rachou - FIOCRUZ-Minas, Belo Horizonte, MG, Brazil.
authors.forename: Olindo Assis
authors.initials: OA
authors.lastname: Martins-Filho
authors.affiliation: Fundação Centro de Hematologia e Hemoterapia de Minas Gerais - HEMOMINAS, Belo Horizonte, MG, Brazil.
mesh_terms: D000465:Algorithms; D058070:Asymptomatic Diseases; D015415:Biomarkers / Q000097:blood; D016022:Case-Control Studies; D002460:Cell Line; D016000:Cluster Analysis; D016207:Cytokines / Q000097:blood; D057225:Data Mining / Q000379:methods; D003663:Decision Trees; D015480:Deltaretrovirus Antibodies / Q000097:blood*; D003937:Diagnosis, Differential; D018450:Disease Progression; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D015490:HTLV-I Infections / Q000097:blood / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D057166:High-Throughput Screening Assays / Q000379:methods*; D015368:Human T-lymphotropic virus 1 / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood*; D015493:Paraparesis, Tropical Spastic / Q000097:blood / Q000175:diagnosis / Q000276:immunology / Q000821:virology; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D013997:Time Factors; D019562:Viral Load
chemical_list: D015415:Biomarkers; D016207:Cytokines; D015480:Deltaretrovirus Antibodies; D007074:Immunoglobulin G
keywords: Clinical follow-up; Diagnosis; Flow cytometry; HAM/TSP; HTLV-1