A New Flow Cytometry-Based Single Platform for Universal and Differential Serodiagnosis of HTLV-1/2 Infection
In the present work, we developed and evaluated the performance of a new flow cytometry-based single platform, referred to as “FC-Duplex IgG1 (HTLV-1/2)”, for universal and differential serodiagnosis of HTLV-1/2 infection. The proposed technology employs a system for detection of IgG1 antibodies in...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-04-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.795815/full |
| _version_ | 1811233662501388288 |
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| author | Luciene Pimenta de Paiva Jordana Grazziela Alves Coelho-dos-Reis Jordana Grazziela Alves Coelho-dos-Reis Bruno Caetano Trindade Vanessa Peruhype-Magalhães Márcio Sobreira Silva Araújo Juan Jonathan Gonçalves Ana Caroline Nogueira-Souza Júlia Pereira Martins Ágata Lopes Ribeiro Ágata Lopes Ribeiro Ana Lucia Starling Luiz Carlos Júnior Alcântara Maísa Aparecida Ribeiro Anna Bárbara de Freitas Carneiro-Proietti Ester Cerdeira Sabino Kelly Alves Bicalho Andréa Teixeira-Carvalho Olindo Assis Martins-Filho |
| author_facet | Luciene Pimenta de Paiva Jordana Grazziela Alves Coelho-dos-Reis Jordana Grazziela Alves Coelho-dos-Reis Bruno Caetano Trindade Vanessa Peruhype-Magalhães Márcio Sobreira Silva Araújo Juan Jonathan Gonçalves Ana Caroline Nogueira-Souza Júlia Pereira Martins Ágata Lopes Ribeiro Ágata Lopes Ribeiro Ana Lucia Starling Luiz Carlos Júnior Alcântara Maísa Aparecida Ribeiro Anna Bárbara de Freitas Carneiro-Proietti Ester Cerdeira Sabino Kelly Alves Bicalho Andréa Teixeira-Carvalho Olindo Assis Martins-Filho |
| author_sort | Luciene Pimenta de Paiva |
| collection | DOAJ |
| description | In the present work, we developed and evaluated the performance of a new flow cytometry-based single platform, referred to as “FC-Duplex IgG1 (HTLV-1/2)”, for universal and differential serodiagnosis of HTLV-1/2 infection. The proposed technology employs a system for detection of IgG1 antibodies in a single competitive immunofluorescence platform by flow cytometry using fluorescently labeled MT-2/MoT cell line mix coupled to a highly sensitive development system (Biotin/Streptavidin/Phycoerythrin). The stability of fluorescent labeling and the antigenicity of MT-2 and MoT cell lines were confirmed upon storage at −20°C for 2, 6, and 12 months. The anti-HTLV-1/2 IgG1 reactivity, expressed as percentage of positive fluorescent cells (PPFC), was evaluated for each target antigen along the titration curve of test serum samples (1:32 to 1:4,096). Upon selection of target cell line and serum dilutions with higher segregation score between groups, the performance of “FIX” and “FIX & PERM” protocols was evaluated. The “FIX” protocol presented excellent performance indices (Se = 92%/Sp = 94%/AUC = 0.96; Se = 96%/Sp = 100%/AUC = 0.99) for the universal (HTLV-1/2 vs. NI) and differential (HTLV-1 vs. HTLV-2) diagnosis of HTLV-1 infection, respectively. Optimization of the “FIX” protocol using the principle of synchronous and asynchronous pairwise analysis further improved the performance of “FC-Duplex IgG1 (HTLV-1/2)”, using the “FIX” protocol for differential diagnosis of HTLV-1 and HTLV-2 infections (Se = 100%/Sp = 100%/AUC = 1.00). In conclusion, the “FC-Duplex IgG1 (HTLV-1/2)” method represents an innovation in the biotechnology segment with the potential to compose a serological kit for differential diagnosis of HTLV-1/2 infection for reference laboratories and blood centers. |
| first_indexed | 2024-04-12T11:23:21Z |
| format | Article |
| id | doaj.art-71200c85c29941dd85e7f0b10483db5a |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-12T11:23:21Z |
| publishDate | 2022-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-71200c85c29941dd85e7f0b10483db5a2022-12-22T03:35:18ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-04-011310.3389/fimmu.2022.795815795815A New Flow Cytometry-Based Single Platform for Universal and Differential Serodiagnosis of HTLV-1/2 InfectionLuciene Pimenta de Paiva0Jordana Grazziela Alves Coelho-dos-Reis1Jordana Grazziela Alves Coelho-dos-Reis2Bruno Caetano Trindade3Vanessa Peruhype-Magalhães4Márcio Sobreira Silva Araújo5Juan Jonathan Gonçalves6Ana Caroline Nogueira-Souza7Júlia Pereira Martins8Ágata Lopes Ribeiro9Ágata Lopes Ribeiro10Ana Lucia Starling11Luiz Carlos Júnior Alcântara12Maísa Aparecida Ribeiro13Anna Bárbara de Freitas Carneiro-Proietti14Ester Cerdeira Sabino15Kelly Alves Bicalho16Andréa Teixeira-Carvalho17Olindo Assis Martins-Filho18Instituto René Rachou, Fundação Oswaldo Cruz (FIOCRUZ)-Minas, Belo Horizonte, BrazilInstituto René Rachou, Fundação Oswaldo Cruz (FIOCRUZ)-Minas, Belo Horizonte, BrazilInstituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, BrazilInstituto René Rachou, Fundação Oswaldo Cruz (FIOCRUZ)-Minas, Belo Horizonte, BrazilInstituto René Rachou, Fundação Oswaldo Cruz (FIOCRUZ)-Minas, Belo Horizonte, BrazilInstituto René Rachou, Fundação Oswaldo Cruz (FIOCRUZ)-Minas, Belo Horizonte, BrazilInstituto René Rachou, Fundação Oswaldo Cruz (FIOCRUZ)-Minas, Belo Horizonte, BrazilInstituto René Rachou, Fundação Oswaldo Cruz (FIOCRUZ)-Minas, Belo Horizonte, BrazilInstituto René Rachou, Fundação Oswaldo Cruz (FIOCRUZ)-Minas, Belo Horizonte, BrazilInstituto René Rachou, Fundação Oswaldo Cruz (FIOCRUZ)-Minas, Belo Horizonte, BrazilInstituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, BrazilFaculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, BrazilInstituto Oswaldo Cruz, Fundação Oswaldo Cruz (FIOCRUZ)/RJ, Rio de Janeiro, BrazilFundação Centro de Hematologia e Hemoterapia do Estado de Minas Gerais- Hemominas (HEMOMINAS), Belo Horizonte, BrazilFundação Centro de Hematologia e Hemoterapia do Estado de Minas Gerais- Hemominas (HEMOMINAS), Belo Horizonte, BrazilUniversidade de São Paulo, Ribeirão Preto, BrazilInstituto René Rachou, Fundação Oswaldo Cruz (FIOCRUZ)-Minas, Belo Horizonte, BrazilInstituto René Rachou, Fundação Oswaldo Cruz (FIOCRUZ)-Minas, Belo Horizonte, BrazilInstituto René Rachou, Fundação Oswaldo Cruz (FIOCRUZ)-Minas, Belo Horizonte, BrazilIn the present work, we developed and evaluated the performance of a new flow cytometry-based single platform, referred to as “FC-Duplex IgG1 (HTLV-1/2)”, for universal and differential serodiagnosis of HTLV-1/2 infection. The proposed technology employs a system for detection of IgG1 antibodies in a single competitive immunofluorescence platform by flow cytometry using fluorescently labeled MT-2/MoT cell line mix coupled to a highly sensitive development system (Biotin/Streptavidin/Phycoerythrin). The stability of fluorescent labeling and the antigenicity of MT-2 and MoT cell lines were confirmed upon storage at −20°C for 2, 6, and 12 months. The anti-HTLV-1/2 IgG1 reactivity, expressed as percentage of positive fluorescent cells (PPFC), was evaluated for each target antigen along the titration curve of test serum samples (1:32 to 1:4,096). Upon selection of target cell line and serum dilutions with higher segregation score between groups, the performance of “FIX” and “FIX & PERM” protocols was evaluated. The “FIX” protocol presented excellent performance indices (Se = 92%/Sp = 94%/AUC = 0.96; Se = 96%/Sp = 100%/AUC = 0.99) for the universal (HTLV-1/2 vs. NI) and differential (HTLV-1 vs. HTLV-2) diagnosis of HTLV-1 infection, respectively. Optimization of the “FIX” protocol using the principle of synchronous and asynchronous pairwise analysis further improved the performance of “FC-Duplex IgG1 (HTLV-1/2)”, using the “FIX” protocol for differential diagnosis of HTLV-1 and HTLV-2 infections (Se = 100%/Sp = 100%/AUC = 1.00). In conclusion, the “FC-Duplex IgG1 (HTLV-1/2)” method represents an innovation in the biotechnology segment with the potential to compose a serological kit for differential diagnosis of HTLV-1/2 infection for reference laboratories and blood centers.https://www.frontiersin.org/articles/10.3389/fimmu.2022.795815/fulluniversal and differential diagnosisHTLV-1/2flow cytometrycompetitive assayHTLV |
| spellingShingle | Luciene Pimenta de Paiva Jordana Grazziela Alves Coelho-dos-Reis Jordana Grazziela Alves Coelho-dos-Reis Bruno Caetano Trindade Vanessa Peruhype-Magalhães Márcio Sobreira Silva Araújo Juan Jonathan Gonçalves Ana Caroline Nogueira-Souza Júlia Pereira Martins Ágata Lopes Ribeiro Ágata Lopes Ribeiro Ana Lucia Starling Luiz Carlos Júnior Alcântara Maísa Aparecida Ribeiro Anna Bárbara de Freitas Carneiro-Proietti Ester Cerdeira Sabino Kelly Alves Bicalho Andréa Teixeira-Carvalho Olindo Assis Martins-Filho A New Flow Cytometry-Based Single Platform for Universal and Differential Serodiagnosis of HTLV-1/2 Infection Frontiers in Immunology universal and differential diagnosis HTLV-1/2 flow cytometry competitive assay HTLV |
| title | A New Flow Cytometry-Based Single Platform for Universal and Differential Serodiagnosis of HTLV-1/2 Infection |
| title_full | A New Flow Cytometry-Based Single Platform for Universal and Differential Serodiagnosis of HTLV-1/2 Infection |
| title_fullStr | A New Flow Cytometry-Based Single Platform for Universal and Differential Serodiagnosis of HTLV-1/2 Infection |
| title_full_unstemmed | A New Flow Cytometry-Based Single Platform for Universal and Differential Serodiagnosis of HTLV-1/2 Infection |
| title_short | A New Flow Cytometry-Based Single Platform for Universal and Differential Serodiagnosis of HTLV-1/2 Infection |
| title_sort | new flow cytometry based single platform for universal and differential serodiagnosis of htlv 1 2 infection |
| topic | universal and differential diagnosis HTLV-1/2 flow cytometry competitive assay HTLV |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.795815/full |
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