A comparison of Plasmodium falciparum circumsporozoite protein-based slot blot and ELISA immuno-assays for oocyst detection in mosquito homogenates

Malaria Journal

Research

Will Stone¹  Kjerstin Lanke¹  Teun Bousema²  Bryan Grabias³  Sanjai Kumar³  Hong Zheng³  Diadier Diallo⁴  Ashley Birkett⁴  Merribeth Morin⁴  Emily Locke⁴ 

[1] 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 and Infection, London School of Hygiene and Tropical Medicine, Keppel Street, London, UK;Laboratory of Emerging Pathogens, Division of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Rockville, MD, USA;PATH Malaria Vaccine Initiative, Washington DC, USA;

关键词: Plasmodium falciparum;    Anopheles;    Sporozoite;    Oocyst;    Circumsporozoite protein;    ELISA;    Slot-blot;    Immuno-assay;    Mosquito infection;   

DOI  :  10.1186/s12936-015-0954-2

 received in 2015-07-15, accepted in 2015-10-21,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundThe infectivity of Plasmodium gametocytes is typically determined by microscopically examining the midguts of mosquitoes that have taken a blood meal containing potentially infectious parasites. Such assessments are required for the development and evaluation of transmission-reducing interventions (TRI), but are limited by subjectivity, technical complexity and throughput. The detection of circumsporozoite protein (CSP) by enzyme-linked immunosorbent assay (ELISA) and enhanced chemiluminescent slot-blot (ECL-SB) may be used as objective, scalable alternatives to microscopy for the determination of infection prevalence.MethodsTo compare the performance of the CSP ELISA and ECL-SB for the detection of mosquito infection, four groups of Anopheles stephensi mosquitoes were infected with cultured Plasmodium falciparum gametocytes. At day-8 post-infection (PI), parasite status was determined by microscopy for a sample of mosquitoes from each group. At days 8 and 10 PI, the parasite status of separate mosquito samples was analysed by both CSP ELISA and ECL-SB.ResultsWhen mosquito samples were analysed 8 days PI, the ECL-SB determined similar infection prevalence to microscopy; CSP ELISA lacked the sensitivity to detect CSP in all infected mosquitoes at this early time point. When mosquitoes were analysed 48 h later (10 days PI) both assays performed as well as microscopy for infection detection.ConclusionsWhilst microscopical examination of mosquito guts is of great value when quantification of parasite burden is required, ECL-SB and CSP ELISA are suitable alternatives at day 10 PI when infection prevalence is the desired endpoint, although CSP ELISA is not suitable at day 8 PI. These results are important to groups considering large-scale implementation of TRI.

【 授权许可】

CC BY   
© Stone et al. 2015