Plasmodium falciparum HRP2 ELISA for analysis of dried blood spot samples in rural Zambia.
Plasmodium falciparum HRP2 ELISA for analysis of dried blood spot samples in rural Zambia.
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恶性疟原虫HRP2 ELISA用于分析赞比亚农村地区的干血点样品。
DOI：
10.1186/s12936-017-1996-4
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发表时间：
2017-08-23
期刊：
影响因子：
3
通讯作者：
Wright DW
中科院分区：
文献类型：
--
作者：
Gibson LE;Markwalter CF;Kimmel DW;Mudenda L;Mbambara S;Thuma PE;Wright DW
干血斑通常用于临床和非临床环境中的样本采集。该方法简单，样品中的生物分子在室温下可保持稳定数月。在现场，用于研究和诊断疟疾的血液样本通常收集在干燥的血点卡上，因此需要开发生物标志物提取和分析方法。对从干血斑中提取疟疾生物标志物恶性疟原虫富含组氨酸蛋白 2 (HRP2) 的简单程序进行了优化，以实现最大的提取效率。该方法用于评估干血斑中 HRP2 的稳定性。此外，使用开发的方法对来自赞比亚农村地区的 328 名患者样本进行了 HRP2 分析。这些样本是在首次接受基于青蒿素的联合疗法时以及治疗后的几个时间点收集的。 HRP2 的平均提取效率为 70%，检测限较低。在特定的储存条件下，HRP2 在干燥的血斑中可稳定至少 6 个月。对患者样本的分析表明，与显微镜检查相比，该方法的灵敏度为 94%，特异性为 89%，并且观察到治疗后 HRP2 清除率的趋势。发现 HRP2 的干血斑 ELISA 灵敏、特异且准确。该方法有效地用于评估患者样本中的生物标志物清除特征，这证明它是进一步了解疾病和流行病学应用的理想选择。本文的在线版本 (doi:10.1186/s12936-017-1996-4) 包含补充材料，可供授权用户使用。
Dried blood spots are commonly used for sample collection in clinical and non-clinical settings. This method is simple, and biomolecules in the samples remain stable for months at room temperature. In the field, blood samples for the study and diagnosis of malaria are often collected on dried blood spot cards, so development of a biomarker extraction and analysis method is needed. A simple extraction procedure for the malarial biomarker Plasmodium falciparum histidine-rich protein 2 (HRP2) from dried blood spots was optimized to achieve maximum extraction efficiency. This method was used to assess the stability of HRP2 in dried blood spots. Furthermore, 328 patient samples made available from rural Zambia were analysed for HRP2 using the developed method. These samples were collected at the initial administration of artemisinin-based combination therapy and at several points following treatment. An average extraction efficiency of 70% HRP2 with a low picomolar detection limit was achieved. In specific storage conditions HRP2 was found to be stable in dried blood spots for at least 6 months. Analysis of patient samples showed the method to have a sensitivity of 94% and a specificity of 89% when compared with microscopy, and trends in HRP2 clearance after treatment were observed. The dried blood spot ELISA for HRP2 was found to be sensitive, specific and accurate. The method was effectively used to assess biomarker clearance characteristics in patient samples, which prove it to be ideal for gaining further insight into the disease and epidemiological applications. The online version of this article (doi:10.1186/s12936-017-1996-4) contains supplementary material, which is available to authorized users.
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DOI：
10.1016/j.jpba.2016.06.026
发表时间：
2016-10-25
影响因子：
3.4
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
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