title: High degree of concordance between flow cytometry and geno2pheno methods for HIV-1 tropism determination in proviral DNA.
abstract: Use of CCR5 antagonists requires previous viral tropism determination. The available methods have high cost, are time-consuming, or require highly trained personnel, and sophisticated equipment. We compared a flow cytometry-based tropism assay with geno2pheno method to determine HIV-1 tropism in AIDS patients, in Bahia, Brazil. We tested peripheral blood mononuclear cells of 102 AIDS patients under antiretroviral therapy by using a cytometry-based tropism assay and geno2pheno assay. Cellular membrane receptors were identified by using CXCR4, CCR5 and CD4 monoclonal antibodies, while detection of cytoplasmic mRNAs for gag and pol HIV regions was achieved by using a labeled probe. Genotypic identification of X4 and R5 tropic viruses was attempted by geno2pheno algorithm. There was a high degree of concordance between cytometry-based tropism assay and geno2pheno algorithm in determination of HIV-1 tropism. Cytometry-based tropism assay demonstrated higher sensitivity and specificity in comparison to geno2pheno, which was used as a gold-standard. One sample could not be amplified by geno2pheno method, but was classified as duotropic by cytometry-based tropism assay. We did not find any association between CD4+ count or plasma HIV-1 RNA viral load and tropism results. The overall performances of cytometry-based tropism assay and geno2pheno assay were almost identical in determination of HIV-1 tropism.
authors.affiliation: Laboratório de Pesquisa em Infectologia, Hospital Universitário Professor Edgard Santos, Universidade Federal da Bahia, Salvador, BA, Brazil. Electronic address: ajltorres@gmail.com.
authors.forename: Alex José Leite
authors.initials: AJ
authors.lastname: Torres
authors.affiliation: Grupo de pesquisa para caracterização de Retrovírus em humanos do Instituto Adolfo Lutz, São Paulo, SP, Brazil.
authors.forename: Luis Fernando de Macedo
authors.initials: LF
authors.lastname: Brígido
authors.affiliation: Laboratório de Pesquisa em Infectologia, Hospital Universitário Professor Edgard Santos, Universidade Federal da Bahia, Salvador, BA, Brazil.
authors.forename: Marcos Herculano Nunes
authors.initials: MH
authors.lastname: Abrahão
authors.affiliation: Laboratório de Pesquisa em Infectologia, Hospital Universitário Professor Edgard Santos, Universidade Federal da Bahia, Salvador, BA, Brazil.
authors.forename: Ana Luiza Dias
authors.initials: AL
authors.lastname: Angelo
authors.affiliation: Laboratório de Pesquisa em Infectologia, Hospital Universitário Professor Edgard Santos, Universidade Federal da Bahia, Salvador, BA, Brazil.
authors.forename: Gilcivaldo
authors.initials: G
authors.lastname: de Jesus Ferreira
authors.affiliation: Grupo de pesquisa para caracterização de Retrovírus em humanos do Instituto Adolfo Lutz, São Paulo, SP, Brazil.
authors.forename: Luana Portes
authors.initials: LP
authors.lastname: Coelho
authors.affiliation: Grupo de pesquisa para caracterização de Retrovírus em humanos do Instituto Adolfo Lutz, São Paulo, SP, Brazil.
authors.forename: João Leandro
authors.initials: JL
authors.lastname: Ferreira
authors.affiliation: Laboratório de Pesquisa em Infectologia, Hospital Universitário Professor Edgard Santos, Universidade Federal da Bahia, Salvador, BA, Brazil.
authors.forename: Célia Regina Mayoral Pedroso
authors.initials: CR
authors.lastname: Jorge
authors.affiliation: Laboratório de Pesquisa em Infectologia, Hospital Universitário Professor Edgard Santos, Universidade Federal da Bahia, Salvador, BA, Brazil.
authors.forename: Eduardo Martins
authors.initials: EM
authors.lastname: Netto
authors.affiliation: Laboratório de Pesquisa em Infectologia, Hospital Universitário Professor Edgard Santos, Universidade Federal da Bahia, Salvador, BA, Brazil.
authors.forename: Carlos
authors.initials: C
authors.lastname: Brites
mesh_terms: D000465:Algorithms; D015496:CD4-Positive T-Lymphocytes; D018414:CD8-Positive T-Lymphocytes; D004279:DNA, Viral / Q000235:genetics*; D005434:Flow Cytometry / Q000379:methods; D005838:Genotype; D015658:HIV Infections / Q000188:drug therapy / Q000821:virology*; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D010641:Phenotype; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D019562:Viral Load; D056189:Viral Tropism / Q000235:genetics*
chemical_list: D004279:DNA, Viral
keywords: Flow cytometry; Geno2pheno; HIV-1; Tropism