Jang Ihn Kyung, Aranda Sara, Barney Rebecca, Rashid Andrew, Helwany Muhammad, Rek John C, Arinaitwe Emmanuel, Adrama Harriet, Murphy Maxwell, Imwong Mallika, Proux Stephane, Haohankhunnatham Warat, Ding Xavier C, Nosten François, Greenhouse Bryan, Gamboa Dionicia, Domingo Gonzalo J
Diagnostics Program, PATH, Seattle, WA USA.
Infectious Diseases Research Collaboration, Kampala, Uganda.
J Parasit Dis. 2021 Jun;45(2):479-489. doi: 10.1007/s12639-020-01325-2. Epub 2021 Jan 3.
Dried blood spots (DBS) typically prepared on filter papers are an ideal sample type for malaria surveillance by offering easy and cost-effective methods in terms of sample collection, storage, and transport. The objective of this study was to evaluate the applicability of DBS with a commercial multiplex malaria assay, developed to concurrently measure antigens, histidine-rich protein 2 (HRP2), lactate dehydrogenase (pLDH), and a host inflammatory biomarker, C-reactive protein (CRP), in whole blood. The assay conditions were optimized for DBS, and thermal stability for measurement of antigens and CRP in dried blood were determined. Performance of the multiplex assay on matched DBS and whole blood pellet samples was also evaluated using the clinical samples. The results indicate the acceptable performance in multiplex antigen detection using DBS samples. At cutoff levels for DBS, with a diagnostic specificity with a lower 95% confidence bound > 92%, diagnostic sensitivities against polymerase chain reaction (PCR)-confirmed malaria for HRP2, LDH, LDH, and Pan LDH were 93.5%, 80.4%, 21.3%, and 55.6%, respectively. The half-life of pLDH was significantly less than that of HRP2 in thermal stability studies. Results with DBS samples collected from Peru indicate that the uncontrolled storage conditions of DBS can result in inaccurate reporting for infection with parasites with deletions. With careful consideration that minimizing the unfavorable DBS storage environment is essential for ensuring integrity of heat-labile antigens, DBS samples can be used as an alternative to liquid whole blood to detect with deletions in malaria surveillance.
The online version of this article (10.1007/s12639-020-01325-2) contains supplementary material, which is available to authorized users.
通常在滤纸上制备的干血斑（DBS）是疟疾监测的理想样本类型，因为在样本采集、储存和运输方面提供了简便且经济高效的方法。本研究的目的是评估DBS与一种商业多重疟疾检测方法的适用性，该方法旨在同时测量全血中的抗原、富含组氨酸蛋白2（HRP2）、乳酸脱氢酶（pLDH）和宿主炎症生物标志物C反应蛋白（CRP）。对DBS的检测条件进行了优化，并确定了干血中抗原和CRP测量的热稳定性。还使用临床样本评估了多重检测在匹配的DBS和全血沉淀样本上的性能。结果表明，使用DBS样本进行多重抗原检测具有可接受的性能。在DBS的临界值水平下，诊断特异性的下限95%置信区间>92%，针对聚合酶链反应（PCR）确诊疟疾的HRP2、LDH、LDH和泛LDH的诊断敏感性分别为93.5%、80.4%、21.3%和55.6%。在热稳定性研究中，pLDH的半衰期明显短于HRP2。从秘鲁采集的DBS样本结果表明，DBS的未控制储存条件可能导致对有缺失寄生虫感染的报告不准确。考虑到尽量减少不利的DBS储存环境对于确保热不稳定抗原的完整性至关重要，DBS样本可作为液体全血的替代品用于疟疾监测中检测有缺失的情况。
本文的在线版本（10.1007/s12639-020-01325-2）包含补充材料，授权用户可获取。
