Plasmodium falciparum HRP2 ELISA for analysis of dried blood spot samples in rural Zambia

Malaria Journal

Research

Lwiindi Mudenda¹  Danielle W. Kimmel¹  David W. Wright¹  Christine F. Markwalter¹  Lauren E. Gibson²  Saidon Mbambara³  Philip E. Thuma³ 

[1] Department of Chemistry, Vanderbilt University, 37235, Nashville, TN, USA;Department of Chemistry, Vanderbilt University, 37235, Nashville, TN, USA;Department of Chemistry and Biochemistry, Elizabethtown College, 17022, Elizabethtown, PA, USA;Macha Research Trust, Choma, Zambia;

关键词: Dried blood spots;    ELISA;    Malaria;    Plasmodium falciparum;    Biomarker clearance;   

DOI  :  10.1186/s12936-017-1996-4

 received in 2017-04-27, accepted in 2017-08-09,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundDried 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.MethodsA 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.ResultsAn 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.ConclusionsThe 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.

【 授权许可】

CC BY   
© The Author(s) 2017