Comparison of parasite load by qPCR and histopathological changes of inner and outer edge of ulcerated cutaneous lesions of cutaneous leishmaniasis

Cutaneous leishmaniasis (CL) is an infectious vector-borne disease caused by protozoa of the Leishmania genus that affects humans and animals. The distribution of parasites in the lesion is not uniform, and there are divergences in the literature about the choice of the better sampling site for diag...

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Bibliographic Details
Published in:PloS one 2021-01, Vol.16 (1), p.e0243978-e0243978
Main Authors: Thomaz, Caio, de Mello, Cintia Xavier, Espíndola, Otávio de Melo, Shubach, Armando de Oliveira, Quintella, Leonardo Pereira, de Oliveira, Raquel Vasconcelos Carvalhaes, Duarte, Adriane Corrêa Gomes, Pimentel, Maria Inês Fernandes, Lyra, Marcelo Rosandiski, Marzochi, Mauro Celio de Almeida
Format: Article
Language:English
Subjects:
Adult
Biology and Life Sciences
Biopsy
Comparative analysis
Cutaneous leishmaniasis
Deoxyribonucleic acid
Diagnosis
DNA
DNA, Kinetoplast - analysis
Endopeptidase K
Female
Fluorimetry
Fluorometers
Forensic science
Histology, Pathological
Humans
Infectious diseases
Laboratories
Leishmania braziliensis - genetics
Leishmania braziliensis - isolation & purification
Leishmaniasis, Cutaneous
Leishmaniasis, Cutaneous - parasitology
Lesions
Load
Lysis
Male
Medicine and Health Sciences
Microscopy
Middle Aged
Parasite Load
Parasites
Parasitic diseases
Polymerase chain reaction
Proteinase
Protozoa
Reagents
Real-Time Polymerase Chain Reaction - methods
Skin diseases
Skin lesions
Surveillance
Ulcer - parasitology
Ulcers
Vector-borne diseases
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Summary:Cutaneous leishmaniasis (CL) is an infectious vector-borne disease caused by protozoa of the Leishmania genus that affects humans and animals. The distribution of parasites in the lesion is not uniform, and there are divergences in the literature about the choice of the better sampling site for diagnosis-inner or outer edge of the ulcerated skin lesion. In this context, determining the region of the lesion with the highest parasite density and, consequently, the appropriate site for collecting samples can define the success of the laboratory diagnosis. Hence, this study aims to comparatively evaluate the parasite load by qPCR, quantification of amastigotes forms in the direct exam, and the histopathological profile on the inner and outer edges of ulcerated CL lesions. Samples from ulcerated skin lesions from 39 patients with confirmed CL were examined. We performed scraping of the ulcer inner edge (base) and outer edge (raised border) and lesion biopsy for imprint and histopathological examination. Slides smears were stained by Giemsa and observed in optical microscopy, the material contained on the smears was used to determine parasite load by quantitative real-time PCR (qPCR) with primers directed to the Leishmania (Viannia) minicircle kinetoplast DNA. The histopathological exam was performed to evaluate cell profile, tissue alterations and semi-quantitative assessment of amastigote forms in inner and outer edges. Parasite loads were higher on the inner edge compared to the outer edge of the lesions, either by qPCR technique (P
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0243978