HRM Accuracy and Limitations as a Species Typing Tool for Leishmania Parasites

HRM 作为利什曼原虫物种分型工具的准确性和局限性

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作者:Camila Patricio Braga Filgueira, Daniela Pitta-Pereira, Lilian Motta Cantanhêde, Gabriel Eduardo Melim Ferreira, Sayonara Dos Reis, Elisa Cupolillo, Otacilio C Moreira, Constança Britto, Mariana Côrtes Boité

Abstract

High Resolution Melting Analysis (HRM) has been pointed out as a suitable alternative method to detect and identify Leishmania species. Herein, we aimed to evaluate the sensitivity, specificity, accuracy, and limitations of a HSP70-HRM protocol both as a diagnostic scheme applied in clinical samples and as a species typing tool for laboratory research and reference services. Our data reveal the pronounced species-typing potential of the HSP70-HRM in DNA from cultured parasites. For clinical samples, however, we advise caution due to parasite load-dependent accuracy. In light of these findings and considering the importance of parasite load determination for clinical and research purposes, we recommend the integration of the presented typing scheme and the previously published Leishmania quantifying approach as combined tools for clinicians, surveillance, and research.

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