Combined DNA extraction and antibody elution from filter papers for the assessment of malaria transmission intensity in epidemiological studies

Malaria Journal

Methodology

Shehu Shagari¹  Euniah Makori¹  Simon Kariuki¹  Victor Osoti¹  Jennifer Stevenson²  Jonathan Cox²  Lynn Grignard³  Colin Sutherland³  Chris Drakeley³  Amrish Baidjoe⁴  Robert Sauerwein⁴  Ivo Ploemen⁴  Will Stone⁴  Teun Bousema⁵ 

[1] Centre for Global Health Research, Kenya Medical Research Institute and Centers of Disease control and prevention, Kisumu, Kenya;Department of Disease Control, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, WC1E 7HT, London, UK;Department of Immunology & Infection, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, WC1E 7HT, London, UK;Department of Medical Microbiology, Radboud University Nijmegen Medical Centre, Nijmegen, the Netherlands;Department of Medical Microbiology, Radboud University Nijmegen Medical Centre, Nijmegen, the Netherlands;Department of Immunology & Infection, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, WC1E 7HT, London, UK;

关键词: Plasmodium;    Malaria;    Antibodies;    IgG;    DNA;    Submicroscopic;    Extraction;    Transmission;    PCR;    ELISA;   

DOI  :  10.1186/1475-2875-12-272

 received in 2013-02-17, accepted in 2013-06-30,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundInforming and evaluating malaria control efforts relies on knowledge of local transmission dynamics. Serological and molecular tools have demonstrated great sensitivity to quantify transmission intensity in low endemic settings where the sensitivity of traditional methods is limited. Filter paper blood spots are commonly used a source of both DNA and antibodies. To enhance the operational practicability of malaria surveys, a method is presented for combined DNA extraction and antibody elution.MethodsFilter paper blood spots were collected as part of a large cross-sectional survey in the Kenyan highlands. DNA was extracted using a saponin/chelex method. The eluate of the first wash during the DNA extraction process was used for antibody detection and compared with previously validated antibody elution procedures. Antibody elution efficiency was assessed by total IgG ELISA for malaria antigens apical membrane antigen-1 (AMA-1) and merozoite-surface protein-1 (MSP-142). The sensitivity of nested 18S rRNA and cytochrome b PCR assays and the impact of doubling filter paper material for PCR sensitivity were determined. The distribution of cell material and antibodies throughout filter paper blood spots were examined using luminescent and fluorescent reporter assays.ResultsAntibody levels measured after the combined antibody/DNA extraction technique were strongly correlated to those measured after standard antibody elution (p < 0.0001). Antibody levels for both AMA-1 and MSP-142 were generally slightly lower (11.3-21.4%) but age-seroprevalence patterns were indistinguishable. The proportion of parasite positive samples ranged from 12.9% to 19.2% in the different PCR assays. Despite strong agreement between outcomes of different PCR assays, none of the assays detected all parasite-positive individuals. For all assays doubling filter paper material for DNA extraction increased sensitivity. The concentration of cell and antibody material was not homogenously distributed throughout blood spots.ConclusionCombined DNA extraction and antibody elution is an operationally attractive approach for high throughput assessment of cumulative malaria exposure and current infection prevalence in endemic settings. Estimates of antibody prevalence are unaffected by the combined extraction and elution procedure. The choice of target gene and the amount and source of filter paper material for DNA extraction can have a marked impact on PCR sensitivity.

【 授权许可】

Unknown   
© Baidjoe et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.