基于非侵入性技术提取样本的疟疾诊断方法:护理临床实践的机会。

Method for Malaria Diagnosis Based on Extractions of Samples Using Non-Invasive Techniques: An Opportunity for the Nursing Clinical Practice.

机构信息

Department of Nursing, Nursing and Occupational Therapy College, University of Extremadura s/n, 10003 Cáceres, Spain.

Unidad de Genética, Facultad de Veterinaria, University of Extremadura s/n, 10003 Cáceres, Spain.

出版信息

Int J Environ Res Public Health. 2020 Jul 31;17(15):5551. doi: 10.3390/ijerph17155551.

Abstract

Malaria has been for millennia one of the best known and most destructive diseases affecting humans. Its high impact has aroused great interest for the development of new effective and reliable diagnostic techniques. Recently it has been recently published that hairs from mammal hosts are able to capture, hold and finally remove foreign DNA sequences of parasites. The aim of this study was to check if () DNA remains stable in blood samples deposited in Whatman paper after suffering different transport and storage conditions, and to compare the sensitivity of these results with those offered by thick a smear and Rapid Diagnostic Test, and besides to examine whether DNA would be detected and quantified by Real time quantitative PCR (qPCR) from hairs of people with different types of malaria. Histidine Repeat Protein II (pHRP-II) antigen detection and DNA were detected in 18 of 19 dry blood samples adhered to Whatman paper (94.74%), besides, DNA was also detected in seven out of 19 hair samples analyzed (36.84%), remaining stable until analysis for several months under the exposure to different environmental conditions. Although the sensitivity of PCR for the diagnosis of malaria in hair samples is not as high as blood analysis, the study of DNA presence in blood and hair could constitute a complementary tool with numerous advantages in sample collection, transport and storage. We suggest that the method could be also applied to medical, forensic and paleo-parasitological diagnosis, not only for malaria but also for searching many other pathogens in hair samples.

摘要

疟疾是几千年来最著名和最具破坏性的人类疾病之一。它的高影响力引起了人们极大的兴趣,促使人们开发新的有效和可靠的诊断技术。最近有研究表明,哺乳动物的毛发能够捕获、固定并最终去除寄生虫的外源 DNA 序列。本研究旨在检测在经过不同的运输和储存条件后,储存于 Whatman 滤纸上的血液样本中的 () DNA 是否稳定,并将这些结果的灵敏度与厚涂片和快速诊断检测提供的结果进行比较,此外,还要检查实时定量 PCR(qPCR) 是否可以从不同类型疟疾患者的毛发中检测和定量 () DNA。在 19 个附着于 Whatman 滤纸上的干燥血样中,有 18 个(94.74%)检测到组氨酸重复蛋白 II(pHRP-II)抗原和 DNA,此外,在分析的 19 个毛发样本中有 7 个(36.84%)检测到 DNA,在暴露于不同环境条件下,几个月后仍保持稳定。尽管 PCR 诊断毛发中疟疾的灵敏度不如血液分析高,但研究血液和毛发中 () DNA 的存在可能构成一种具有样本收集、运输和储存等众多优势的补充工具。我们建议该方法也可应用于医学、法医学和古寄生虫学诊断,不仅用于疟疾,还可用于在毛发样本中寻找其他许多病原体。

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