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作为寄生虫物种鉴定工具的 HRM 准确性和局限性。

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

机构信息

Leishmaniasis Research Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro 21040-360, Brazil.

Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz, FIOCRUZ, Pavilhão Leônidas Deane, Sala 209, Avenida Brasil 4365, Manguinhos, Rio de Janeiro 21045-900, Brazil.

出版信息

Int J Mol Sci. 2023 Sep 30;24(19):14784. doi: 10.3390/ijms241914784.

Abstract

High Resolution Melting Analysis (HRM) has been pointed out as a suitable alternative method to detect and identify 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 quantifying approach as combined tools for clinicians, surveillance, and research.

摘要

高分辨率熔解分析(HRM)已被指出是一种合适的替代方法,可用于检测和鉴定物种。在此,我们旨在评估 HSP70-HRM 方案作为临床样本应用的诊断方案以及实验室研究和参考服务的物种分型工具的灵敏度、特异性、准确性和局限性。我们的数据揭示了 HSP70-HRM 在培养寄生虫 DNA 中具有明显的物种分型潜力。然而,对于临床样本,由于寄生虫负荷依赖性准确性,我们建议谨慎使用。鉴于这些发现,并考虑到寄生虫负荷测定对于临床和研究目的的重要性,我们建议将所提出的分型方案和之前发表的定量方法整合为临床医生、监测和研究的联合工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55ee/10573552/fea48e201440/ijms-24-14784-g001.jpg

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