HIRO GOTO
Projetos de Pesquisa
Unidades Organizacionais
Departamento de Medicina Preventiva, Faculdade de Medicina - Docente
LIM/38 - Laboratório de Epidemiologia e Imunobiologia, Hospital das Clínicas, Faculdade de Medicina - Líder
LIM/38 - Laboratório de Epidemiologia e Imunobiologia, Hospital das Clínicas, Faculdade de Medicina - Líder
11 resultados
Resultados de Busca
Agora exibindo 1 - 10 de 11
- Use of Recombinant Antigens for Sensitive Serodiagnosis of American Tegumentary Leishmaniasis Caused by Different Leishmania Species(2017) SATO, Camila Massae; SANCHEZ, Maria Carmen Arroyo; CELESTE, Beatriz Julieta; DUTHIE, Malcolm S.; GUDERIAN, Jeffrey; REED, Steven G.; BRITO, Maria Edileuza Felinto de; CAMPOS, Marliane Batista; ENCARNACAO, Helia Valeria de Souza; GUERRA, Jorge; MESQUITA, Tirza Gabrielle Ramos de; PINHEIRO, Suzana Kanawati; RAMASAWMY, Rajendranath; SILVEIRA, Fernando Tobias; SOUZA, Marina de Assis; GOTO, HiroAmerican tegumentary leishmaniasis (ATL) (also known as cutaneous leishmaniasis [CL]) is caused by various species of protozoa of the genus Leishmania. The diagnosis is achieved on a clinical, epidemiological, and pathological basis, supported by positive parasitological exams and demonstration of leishmanin delayed-type hypersensitivity. Serological assays are not routinely used in the diagnosis because many are considered to have low sensitivity and the particular Leishmania species causing the disease can lead to variable performance. In the present study, we generated recombinant versions of two highly conserved Leishmania proteins, Leishmania (Viannia) braziliensis-derived Lb8E and Lb6H, and evaluated both in enzyme-linked immunosorbent assays (ELISA). Recombinant Lb6H (rLb6H) had better performance and reacted with 100.0% of the ATL and 89.4% of the VL samples. These reactions with rLb6H were highly specific (98.5%) when compared against those for samples from healthy control individuals. We then assessed rLb6H against sera from ATL patients infected with different species of Leishmania prevalent in Brazil [Leishmania (Leishmania) amazonensis, L. (Viannia) braziliensis, and L. (V.) guyanensis] and samples from patients with other infectious diseases. In analyses of 500 sera, ELISA using rLb6H detected all 219 ATL samples (sensitivity of 100.0%) with an overall specificity of 93.9% (considering healthy individuals and other infectious diseases patients). Only a minority of samples from Chagas disease patients possessed antibodies against rLb6H, and all of these responses were low (with a highest reactivity index of 2.2). Taken together, our data support further evaluation of rLb6H and the potential for its routine use in the serological diagnosis of ATL.
- Recombinant Leishmania infantum Heat Shock Protein 83 for the Serodiagnosis of Cutaneous, Mucosal, and Visceral Leishmaniases(2014) CELESTE, Beatriz Julieta; SANCHEZ, Maria Carmen Arroyo; RAMOS-SANCHEZ, Eduardo Milton; CASTRO, Luiz Guilherme M.; COSTA, Francisco Assis Lima; GOTO, HiroRoutine serological diagnoses for leishmaniases, except in visceral cases, are performed using whole-parasite antigens. We used enzyme-linked immunosorbent assay (ELISA) to evaluate the performance of Leishmania infantum rHsp83 compared with L. major-like total promastigote antigen in the diagnosis of cutaneous (CL), mucosal (ML), and visceral leishmaniases (VL). ELISA-rHsp83 was significantly more sensitive than ELISA-L. major-like when considering either CL/ML (P = 0.041) or all leishmaniasis patients (P = 0.013). When samples from other infectious disease patients were evaluated for cross-reactivity, ELISA-rHsp83 was more specific than ELISA-L. major-like, specifically for Chagas disease samples (P < 0.001). We also evaluated the anti-rHsp83 antibody titers months after treatment and observed no significant difference in ML (P = 0.607) or CL (P = 0.205). We recommend ELISA-L. infantum-rHsp83 as a routine confirmatory serological assay for the diagnosis of Leishmania infection because of the high sensitivity, the specificity, and the insignificant cross-reactivity with other infectious diseases.
- Validation of ELISA with recombinant antigens in serological diagnosis of canine Leishmania infantum infection(2021) FUJIMORI, Mahyumi; ALMEIDA, Arleana do Bom Parto Ferreira de; BARROUIN-MELO, Stella Maria; CORTEZ, Luiz Ricardo Paes de Barros; DUTHIE, Malcolm Scott; HIRAMOTO, Roberto Mitsuyoshi; PINHO, Flaviane Alves de; REED, Steven Gregory; SOUSA, Valeria Regia Franco; SOUZA, Nazare Fonseca; SOARES, Rodrigo Martins; TOLEZANO, Jose Eduardo; SANCHEZ, Maria Carmen Arroyo; GOTO, HiroBACKGROUND 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.
conferenceObject Evaluation of immunochromatographic assay with recombinant antigen K39, using whole blood, serum and oral fluid, in the diagnosis of visceral leishmaniasis in Brazilian endemic areas(2015) CELESTE, B. J.; SANCHEZ, M. C. A.; ALMEIDA, R. P. D.; BRANCO, C. M. F. Castelo; LINDOSO, J. A. L.; PANIAGO, A. M. M.; QUEIROZ, I. T.; FUJIMORI, M.; CARAMELO, L. D. C.; GOTO, H.conferenceObject ANTI-LEISHMANIA ANTIBODIES IN SAMPLES OF BLOOD DONORS FROM ENDEMIC AREAS OF BRAZIL(2017) FUJIMORI, Mahyumi; CARAMELO, Luiza da Cruz; DORNELAS, Marina Portanova; CELESTE, Beatriz Julieta; WENDEL, Silvano; GOTO, Hiro; SANCHEZ, Maria Carmen- Leishmania infection in blood donors: A new challenge in leishmaniasis transmission?(2018) FRANCA, Adriana de Oliveira; POMPILIO, Mauricio Antonio; PONTES, Elenir Rose Jardim Cury; OLIVEIRA, Marcia Pereira de; PEREIRA, Luiza Oliveira Ramos; LIMA, Rosimar Baptista; GOTO, Hiro; SANCHEZ, Maria Carmen Arroyo; FUJIMORI, Mahyumi; LIMA-JUNIORS, Manoel Sebastiao da Costa; MATOS, Maria de Fatima Cepa; DORVAL, Maria Elizabeth Moraes CavalheirosTransfusion-transmitted leishmaniasis has been a concern in regions endemic for the disease. Whether immediate or delayed, the risks posed by this mode of transmission call for careful assessment. The purpose of this study was to detect Leishmania infection in blood donors living in an endemic area and to investigate progression to the disease in these individuals. Immunofluorescent antibody test, enzyme-linked immunosorbent assay, leishmaniasis rapid test, and the polymerase chain reaction were applied to 430 donors in an initial evaluation. Of those donors with at least one positive test, 50 were reevaluated four years later by the same methods, as were 25 controls who had been negative on the same tests. In the first evaluation, Leishmania infection was detected in 41.4% (95% CI: 36.7-46.1) of donors (n = 430). None of the 75 reevaluated individuals had developed the disease, but retesting revealed positivity in at least one test in 36.0% (95% CI: 25.1-46.9) of donors. Of the 50 initially testing positive, 50% remained so on retesting. Of the 25 initially negative controls, two tested positive in the subsequent evaluation. The severity of the parasitosis and the risk of transfusion transmission warrant investigation of the potential inclusion of methods for Leishmania detection into blood banks for effective screening of infected donors.
- Recombinant protein KR95 as an alternative for serological diagnosis of human visceral leishmaniasis in the Americas(2023) FUJIMORI, Mahyumi; VALENCIA-PORTILLO, Ruth Tamara; LINDOSO, Jose Angelo Lauletta; CELESTE, Beatriz Julieta; ALMEIDA, Roque Pacheco de; COSTA, Carlos Henrique Nery; CRUZ, Alda Maria da; DRUZIAN, Angelita Fernandes; DUTHIE, Malcolm Scott; FORTALEZA, Carlos Magno Castelo Branco; OLIVEIRA, Ana Lucia Lyrio de; PANIAGO, Anamaria Mello C. Miranda; QUEIROZ, Igor Thiago; REED, Steve; VALLUR, Aarthy; GOTO, Hiro; SANCHEZ, Maria Carmen ArroyoIn the Americas, visceral leishmaniasis (VL) is caused by the protozoan Leishmania infantum, leading to death if not promptly diagnosed and treated. In Brazil, the disease reaches all regions, and in 2020, 1,933 VL cases were reported with 9.5% lethality. Thus, an accurate diagnosis is essential to provide the appropriate treatment. Serological VL diagnosis is based mainly on immunochromatographic tests, but their performance may vary by location, and evaluation of diagnostic alternatives is necessary. In this study, we aimed to evaluate the performance of ELISA with the scantily studied recombinant antigens, K18 and KR95, comparing their performance with the already known rK28 and rK39. Sera from parasitologically confirmed symptomatic VL patients (n = 90) and healthy endemic controls (n = 90) were submitted to ELISA with rK18 and rKR95. Sensitivity (95% CI) was, respectively, 83.3% (74.2-89.7) and 95.6% (88.8-98.6), and specificity (95% CI) was 93.3% (85.9-97.2) and 97.8% (91.8-99.9). For validation of ELISA with the recombinant antigens, we included samples from 122 VL patients and 83 healthy controls collected in three regions in Brazil (Northeast, Southeast, and Midwest). When comparing the results obtained with the VL patients' samples, significantly lower sensitivity was obtained by rK18-ELISA (88.5%, 95% CI: 81.5-93.2) compared with rK28-ELISA (95.9%, 95% CI: 90.5-98.5), but the sensitivity was similar comparing rKR95-ELISA (95.1%, 95% CI: 89.5-98.0), rK28-ELISA (95.9%, 95% CI: 90.5-98.5), and rK39-ELISA (94.3%, 95% CI: 88.4-97.4). Analyzing the specificity, it was lowest with rK18-ELISA (62.7%, 95% CI: 51.9-72.3) with 83 healthy control samples. Conversely, higher and similar specificity was obtained by rKR95-ELISA (96.4%, 95% CI: 89.5-99.2), rK28-ELISA (95.2%, 95% CI: 87.9-98.5), and rK39-ELISA (95.2%, 95% CI: 87.9-98.5). There was no difference in sensitivity and specificity across localities. Cross-reactivity assessment, performed with sera of patients diagnosed with inflammatory disorders and other infectious diseases, was 34.2% with rK18-ELISA and 3.1% with rKR95-ELISA. Based on these data, we suggest using recombinant antigen KR95 in serological assays for VL diagnosis.
- Improved Performance of ELISA and Immunochromatographic Tests Using a New Chimeric A2-Based Protein for Human Visceral Leishmaniasis Diagnosis(2021) FIGUEIREDO, Maria Marta; SANTOS, Anna R. R. dos; GODOI, Lara C.; CASTRO, Natalia S. de; ANDRADE, Bruno C. de; SERGIO, Sarah A. R.; JERONIMO, Selma M. B.; OLIVEIRA, Edward J. de; VALENCIA-PORTILLO, Ruth T.; BEZERRA, Lucilandia M.; GOTO, Hiro; SANCHEZ, Maria C. A.; JUNQUEIRA, Caroline; TEIXEIRA, Santuza M. R.; FONSECA, Flavio G. da; GAZZINELLI, Ricardo T.; FERNANDES, Ana PaulaHuman visceral leishmaniasis (VL) is a major public health problem worldwide, leading to significant mortality rates if not properly treated and controlled. Precise identification of infected patients is essential to establish treatment and control measures. Although several VL serological diagnosis advances have been accomplished lately, mainly using recombinant antigens and immunochromatographic tests (ICTs), improvements may still be achieved using multiepitope chimeric proteins in different test platforms. Here, we reported on the evaluation of ELISA and an ICT developed with a new chimeric protein, named DTL-4, based on repetitive antigenic sequences, including those present in the A2 protein. Methods. A total of 1028 sera samples were used for the development and validation of ELISA (321 samples from L. infantum-infected patients, 62 samples from VL/AIDS coinfected patients, 236 samples from patients infected with other diseases, and 409 samples from healthy donors). A total of 520 sera samples were used to develop and validate ICT (249 samples from L. infantum-infected patients, 46 samples from VL/AIDS coinfected patients, 40 samples from patients infected with other diseases, and 185 samples from healthy donors). Findings. Using the validation sera panels, DTL-4-based ELISA displayed an overall sensitivity of 94.61% (95% CI: 89.94-97.28), a specificity of 99.41% (95% CI: 96.39-99.99), and an accuracy of 97.02% (95% CI: 94.61-98.38), while for ICT, sensitivity, specificity, and accuracy values corresponded to 91.98% (95% CI: 86.65-95.39), 100.00% (95% CI: 96.30-100.00), and 95.14% (95% CI: 91.62-97.15), respectively. When testing sera samples from VL/AIDS coinfected patients, DTL-4-ELISA displayed a sensitivity of 77.42% (95% CI: 65.48-86.16), a specificity of 99.41% (95% CI: 96.39-99.99), and an accuracy of 93.51% (95% CI: 89.49%-96.10%), while for DTL-4-ICT, sensitivity was 73.91% (95% CI: 59.74-84.40), specificity was 90.63% (95% CI: 81.02-95.63), and accuracy was 82.00% (95% CI: 73.63-90.91). Conclusion. DTL-4 is a promising candidate antigen for serodiagnosis of VL patients, including those with VL/AIDS coinfection, when incorporated into ELISA or ICT test formats.
conferenceObject Anti-Leishmania antibodies in samples of blood donors from endemic areas of Brazil using ELISA with recombinant protein K39 of L. infantum(2015) SANCHEZ, M. C. A.; FUJIMORI, M.; CARAMELO, L. D. C.; DORNELAS, M. P.; WENDEL, S.; GOTO, H.- Performance of rK39-based immunochromatographic rapid diagnostic test for serodiagnosis of visceral leishmaniasis using whole blood, serum and oral fluid(2020) SANCHEZ, Maria Carmen Arroyo; CELESTE, Beatriz Julieta; LINDOSO, Jose Angelo Lauletta; FUJIMORI, Mahyumi; ALMEIDA, Roque Pacheco de; FORTALEZA, Carlos Magno Castelo Branco; DRUZIAN, Angelita Fernandes; LEMOS, Ana Priscila Freitas; MELO, Vanessa Campos Andrade de; PANIAGO, Anamaria Mello Miranda; QUEIROZ, Igor Thiago; GOTO, HiroBackground The development of rK39-based immunochromatographic rapid diagnostic tests represents an important advance for serodiagnosis of visceral leishmaniasis, being cheap and easy to use at the point of care (POC). Although the use of rK39 have considerably improved the sensitivity and specificity of serological tests compared with total antigens, great variability in sensitivity and specificity was reported. This study aimed at the evaluation of ""Kalazar Detect (TM) Rapid Test, Whole Blood"" (Kalazar Detect RDT) for Visceral Leishmaniasis (VL) diagnosis using oral fluid, whole blood and serum specimens collected at different endemic areas of VL of Brazil. Methodology To evaluate Kalazar Detect RDT, oral fluid, whole blood and serum specimens from 128 VL patients, 85 healthy individuals, 22 patients with possible cross-reactivity diseases and 20 VL/aids coinfected patients were collected and assayed at the POC. Principal findings and conclusions The performance of Kalazar Detect RDT in whole blood and serum was similar; however, using oral fluid, the sensitivity was low. Particularly in samples from the city of Natal, Rio Grande do Norte state in Northeastern Brazil, we observed low sensitivity, 80.0% (95% CI: 62.7-90.5), using whole blood and serum, and poor sensitivity, 43.3% (95% CI: 27.4-60.8) with oral fluid. Those values were much lower than in the other regions, where sensitivity ranged from 92.7-96.3% in whole blood and serum, and 80.0-88.9% in oral fluid. Besides, in VL/aids coinfected patients, lower sensitivity was achieved compared with VL patients. In samples from Natal, the sensitivity was 0.0% (95% CI: 0.0-49.0) and 25.0% (95% CI: 4.6-69.9), using oral fluid and serum/whole blood, respectively; in samples from the other regions, the sensitivity ranged from 40.0-63.6% and 80.0-81.8%, respectively. As for specificity, high values were observed across the fluids, 100.0% (95% CI: 96.5-100.0) in whole blood, 96.3% (95% CI: 90.8-98.5) in serum, and 95.3% (95% CI: 89.5-98.0) in oral fluid; across localities, specificity ranged from 85.7-100.0%. Serum samples sent by the collaborating centers to Instituto de Medicina Tropical (n = 250) were tested by Kalazar Detect RDT, Direct Agglutination Test, Indirect immunofluorescence assay, Enzyme-linked immunosorbent assay, and IT-Leish (R) RDT. The regional difference in the performance of rK39-based RDT and lower sensitivity in Leishmania/HIV coinfected patients raise concern on the routine use of these products for the diagnosis of VL.