Mahyumi Fujimori1, Arleana do Bom Parto Ferreira de Almeida2, Stella Maria Barrouin-Melo3, Luiz Ricardo Paes de Barros Cortez4, Malcolm Scott Duthie5, Roberto Mitsuyoshi Hiramoto6, Flaviane Alves de Pinho3, Steven Gregory Reed5, Valéria Régia Franco Sousa2, Nazaré Fonseca Souza7, Rodrigo Martins Soares8, José Eduardo Tolezano6, Maria Carmen Arroyo Sanchez1, Hiro Goto1,9. 1. Instituto de Medicina Tropical da Faculdade de Medicina da Universidade de São Paulo, Laboratório de Soroepidemiologia e Imunobiologia, São Paulo, SP, Brasil. 2. Universidade Federal de Mato Grosso, Faculdade de Medicina Veterinária, Departamento de Clínica Médica Veterinária, Cuiabá, MT, Brasil. 3. Universidade Federal da Bahia, Hospital-Escola de Medicina Veterinária, Laboratório de Infectologia Veterinária, Salvador, BA, Brasil. 4. Secretaria Municipal de Saúde de Bauru, Departamento de Saúde Coletiva - Vigilância Ambiental, Bauru, SP, Brasil. 5. Host Directed Therapeutics, Seattle, WA, United States of America. 6. Instituto Adolfo Lutz, Centro de Parasitologia e Micologia, Núcleo de Parasitoses Sistêmicas, São Paulo, SP, Brasil. 7. Universidade Federal Rural da Amazônia, Instituto da Saúde e Produção Animal, Belém, Pará, Brasil. 8. Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Medicina Veterinária Preventiva e Saúde Animal, Laboratório de Doenças Parasitárias, São Paulo, SP, Brasil. 9. Universidade de São Paulo, Faculdade de Medicina, Departamento de Medicina Preventiva, São Paulo, SP, Brasil.
Abstract
BACKGROUND: Dogs are the main peridomiciliary reservoir of Leishmania infantum thus the correct diagnosis of infection is essential for the control of the transmission and treatment as well. However, the diagnosis is based on serological assays that are not fully effective. OBJECTIVE: We aimed to establish an effective serological assay for the diagnosis of L. infantum infected dogs using Leishmania-derived recombinant antigens. METHODS: Leishmania derived rK39-, rK28-, rKR95-based enzyme-linked immunosorbent assay (ELISA) was standardized using symptomatic and asymptomatic L. infantum-infected dogs. Then 2,530 samples from inquiry in endemic areas for VL were evaluated and the results compared with recommended assays by the Brazilian Ministry of Health (MH algorithm). Further samples from a cohort of 30 dogs were searched. FINDINGS: For rK39-, rK28- and rKR95-ELISA the sensitivity was around 97% and specificity 100%. The positivity of these three ELISA in the inquiry samples was 27-28%, around 10% higher than the assays currently in use. When cohort samples were searched, we observed likely false-negative results (> 65%) with supposedly negative samples that turned positive six months later with the assays in use (MH algorithm). MAIN CONCLUSIONS: For the diagnosis of L. infantum-infected dogs, rK39-based ELISA showed better diagnostic performance than other assays in use in Brazil and worldwide.
BACKGROUND:Dogs are the main peridomiciliary reservoir of Leishmania infantum thus the correct diagnosis of infection is essential for the control of the transmission and treatment as well. However, the diagnosis is based on serological assays that are not fully effective. OBJECTIVE: We aimed to establish an effective serological assay for the diagnosis of L. infantuminfecteddogs using Leishmania-derived recombinant antigens. METHODS:Leishmania derived rK39-, rK28-, rKR95-based enzyme-linked immunosorbent assay (ELISA) was standardized using symptomatic and asymptomatic L. infantum-infecteddogs. Then 2,530 samples from inquiry in endemic areas for VL were evaluated and the results compared with recommended assays by the Brazilian Ministry of Health (MH algorithm). Further samples from a cohort of 30 dogs were searched. FINDINGS: For rK39-, rK28- and rKR95-ELISA the sensitivity was around 97% and specificity 100%. The positivity of these three ELISA in the inquiry samples was 27-28%, around 10% higher than the assays currently in use. When cohort samples were searched, we observed likely false-negative results (> 65%) with supposedly negative samples that turned positive six months later with the assays in use (MH algorithm). MAIN CONCLUSIONS: For the diagnosis of L. infantum-infecteddogs, rK39-based ELISA showed better diagnostic performance than other assays in use in Brazil and worldwide.
Authors: Shabir Banoo; David Bell; Patrick Bossuyt; Alan Herring; David Mabey; Freddie Poole; Peter G Smith; N Sriram; Chansuda Wongsrichanalai; Ralf Linke; Rick O'Brien; Mark Perkins; Jane Cunningham; Precious Matsoso; Carl Michael Nathanson; Piero Olliaro; Rosanna W Peeling; Andy Ramsay Journal: Nat Rev Microbiol Date: 2006-09 Impact factor: 60.633
Authors: Sowmya Pattabhi; Jacqueline Whittle; Raodoh Mohamath; Sayda El-Safi; Garner G Moulton; Jeffrey A Guderian; Danny Colombara; Asem O Abdoon; Maowia M Mukhtar; Dinesh Mondal; Javan Esfandiari; Shailendra Kumar; Peter Chun; Steven G Reed; Ajay Bhatia Journal: PLoS Negl Trop Dis Date: 2010-09-14