L Sevilha-Santos1, A C M Dos Santos Júnior2, V Medeiros-Silva3, J O Bergmann1, E F da Silva4, L F Segato5, A Y M Arabi6, N A de Paula7, R N R Sampaio8, B D Lima9, C M Gomes10. 1. Postgraduate Program in Medical Sciences, Faculty of Medicine, University of Brasília (UnB), Brasília, Brazil. 2. Postgraduate Program in Molecular Pathology, Faculty of Medicine, University of Brasília (UnB), Brasília, Brazil. 3. Postgraduate Program in Health Sciences, Faculty of Health Sciences, University of Brasília (UnB), Brasília, Brazil. 4. Department of Statistics, University of Brasília (UnB), Brasília, Brazil. 5. Dermatology Division, University Hospital of Brasília (HUB), University of Brasília (UnB), Brasília, Brazil. 6. Faculty of Medicine, University of Brasília, Brasília, Brazil. 7. Department of Cell and Molecular Biology and Pathogenic Bioagents, Dermatology Division, Department of Medical Clinics, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, Brazil. 8. Postgraduate Program in Medical Sciences, Faculty of Medicine, University of Brasília (UnB), Brasília, Brazil; Postgraduate Program in Health Sciences, Faculty of Health Sciences, University of Brasília (UnB), Brasília, Brazil; Dermatomycology Laboratory, Faculty of Medicine, University of Brasília, Brasília, Brazil. 9. Gene Biology Laboratory, Postgraduate Program in Microbial Biology, Cell Biology Department, Biological Sciences Institute, University of Brasília (UnB), Brasília, DF, Brazil. 10. Postgraduate Program in Medical Sciences, Faculty of Medicine, University of Brasília (UnB), Brasília, Brazil; Dermatology Division, University Hospital of Brasília (HUB), University of Brasília (UnB), Brasília, Brazil; Dermatomycology Laboratory, Faculty of Medicine, University of Brasília, Brasília, Brazil; Tropical Medicine Nucleus, University of Brasília, Brasília, Brazil. Electronic address: ciromgomes@gmail.com.
Abstract
OBJECTIVES: Superficial swab sampling of American tegumentary leishmaniasis (ATL) lesions shows higher amounts of Leishmania than those from biopsy. Subcutaneous involvement is also important in ATL, but parasite quantification according to lesion depth has not been evaluated. We aim to present the best depth at which sampling should be performed for molecular exams of ATL. METHODS: Patients with a clinical presentation compatible with ATL were allocated to ATL and control groups. Qualitative and quantitative qPCR assays were performed using SYBR Green and primers amplifying the kDNA minicircle of Leishmania spp. in different skin layers, including the epidermis, the superior dermis, the inferior dermis, and the hypodermis. RESULTS: Fifty-nine patients were included in this study, including 40 who had been diagnosed with ATL and 19 controls. The number of parasites was greater in samples of the epidermis and superior dermis (159.1 × 106, range 4.0-781.7, and 75.4 × 106, range 8.0-244.5, mean Leishmania parasite equivalents per μg of tissue DNA, respectively) than those in samples of the inferior dermis and hypodermis (54.6, range 8.0-256.6, and 16.8 × 106, range 8.0-24.1, mean Leishmania parasite equivalents per μg of tissue DNA, respectively). The best diagnostic accuracy was achieved in the superior dermis (77.9%) and was significantly greater than that in the hypodermis (63.3%; p 0.039). CONCLUSIONS: We conclude that superficial sampling can retrieve a greater quantity of parasites. Future studies of the role of transepidermal elimination as a mechanism of host defence in ATL must be performed as there is a considerable quantity of Leishmania kDNA in the epidermis.
OBJECTIVES: Superficial swab sampling of American tegumentary leishmaniasis (ATL) lesions shows higher amounts of Leishmania than those from biopsy. Subcutaneous involvement is also important in ATL, but parasite quantification according to lesion depth has not been evaluated. We aim to present the best depth at which sampling should be performed for molecular exams of ATL. METHODS:Patients with a clinical presentation compatible with ATL were allocated to ATL and control groups. Qualitative and quantitative qPCR assays were performed using SYBR Green and primers amplifying the kDNA minicircle of Leishmania spp. in different skin layers, including the epidermis, the superior dermis, the inferior dermis, and the hypodermis. RESULTS: Fifty-nine patients were included in this study, including 40 who had been diagnosed with ATL and 19 controls. The number of parasites was greater in samples of the epidermis and superior dermis (159.1 × 106, range 4.0-781.7, and 75.4 × 106, range 8.0-244.5, mean Leishmania parasite equivalents per μg of tissue DNA, respectively) than those in samples of the inferior dermis and hypodermis (54.6, range 8.0-256.6, and 16.8 × 106, range 8.0-24.1, mean Leishmania parasite equivalents per μg of tissue DNA, respectively). The best diagnostic accuracy was achieved in the superior dermis (77.9%) and was significantly greater than that in the hypodermis (63.3%; p 0.039). CONCLUSIONS: We conclude that superficial sampling can retrieve a greater quantity of parasites. Future studies of the role of transepidermal elimination as a mechanism of host defence in ATL must be performed as there is a considerable quantity of Leishmania kDNA in the epidermis.
Authors: Luciana P Oliveira; Luciana C S Nascimento; Fabiola S Santos; Jaqueline L C Takamatsu; Luiz R P Sanchez; Walter S Santos; Lourdes M Garcez Journal: Int J Environ Res Public Health Date: 2022-05-23 Impact factor: 4.614
Authors: Ana Bárbara Sapienza Pinheiro; Patricia Shu Kurizky; Marina de Freitas Ferreira; Marco Antonio de Souza Mota; Jaqueline Santos Ribeiro; Edson Zuza de Oliveira Filho; Carlos Augusto Souza; Daniel Holanda Barroso; Raimunda Nonata Ribeiro Sampaio; Ciro Martins Gomes Journal: Rev Soc Bras Med Trop Date: 2020-04-27 Impact factor: 1.581
Authors: Lais Sevilha-Santos; Danielle Costa Aquino; Günter Hans Neto; Fabiano José Queiroz Costa; Carlos Augusto Felipe de Sousa; Elaine Faria Morelo; Agenor de Castro Moreira Dos Santos Júnior; Ciro Martins Gomes Journal: Braz J Infect Dis Date: 2022-06-27 Impact factor: 3.257