title: A sensitive enhanced chemiluminescent-ELISA for the detection of Plasmodium falciparum circumsporozoite antigen in midguts of Anopheles stephensi mosquitoes.
abstract: Efforts to develop a successful malaria vaccine are hampered due to lack of assays that are predictive of protective immunity without conducting large clinical studies. The effect of experimental vaccines and drugs on malaria transmission is yet more difficult to measure. Knowledge on the Plasmodium infection rate in mosquito populations will aid the measurement of effects from intervention measures for malaria control. Here, we report the development of a chemiluminescent sandwich ELISA (ECL-ELISA) that can detect Plasmodium falciparum circumsporozoite protein (Pf CSP) produced in recombinant form at concentrations of 4.4pg and in P. falciparum sporozoites (Pf SPZ) derived from mosquito salivary glands at levels corresponding to 5 Pf SPZ. Most importantly, we demonstrate reliable Pf CSP-based detection of 0.056day 8 P. falciparum oocysts developing inside mosquito midguts in whole mosquito lysates. Cumulatively, the ECL-ELISA is 47× more sensitive for the detection of Pf CSP than a colorimetric ELISA while greatly simplifying sample preparation, obviating the need for cumbersome midgut dissections and allowing high throughput screening of Plasmodium infection in mosquito populations. The ECL-ELISA may also have broader application in diagnosis of infectious diseases and the prognostic value in cancer and other diseases such as auto-immunity and genetic disorders based on antigen detection, or quality validation of biological vaccine components.
authors.affiliation: 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. Electronic address: Bryan.Grabias@fda.hhs.gov.
authors.forename: Bryan
authors.initials: B
authors.lastname: Grabias
authors.affiliation: 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. Electronic address: Hong.Zheng@fda.hhs.gov.
authors.forename: Hong
authors.initials: H
authors.lastname: Zheng
authors.affiliation: The Johns Hopkins Malaria Research Institute, Bloomberg School of Public Health, The Johns Hopkins University, Baltimore, MD 21205, United States; Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, The Johns Hopkins University, Baltimore, MD 21205, United States. Electronic address: gmlambo1@jhu.edu.
authors.forename: Godfree
authors.initials: G
authors.lastname: Mlambo
authors.affiliation: The Johns Hopkins Malaria Research Institute, Bloomberg School of Public Health, The Johns Hopkins University, Baltimore, MD 21205, United States; Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, The Johns Hopkins University, Baltimore, MD 21205, United States. Electronic address: atripat2@jhu.edu.
authors.forename: Abhai K
authors.initials: AK
authors.lastname: Tripathi
authors.affiliation: 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. Electronic address: Sanjai.Kumar@fda.hhs.gov.
authors.forename: Sanjai
authors.initials: S
authors.lastname: Kumar
mesh_terms: D000818:Animals; D000852:Anopheles / Q000737:chemistry / Q000469:parasitology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D041981:Gastrointestinal Tract / Q000737:chemistry / Q000469:parasitology; D007303:Insect Vectors / Q000737:chemistry / Q000469:parasitology; D008297:Male; D008807:Mice, Inbred BALB C; D010963:Plasmodium falciparum / Q000737:chemistry* / Q000254:growth & development / Q000302:isolation & purification / Q000378:metabolism; D015800:Protozoan Proteins / Q000032:analysis* / Q000378:metabolism; D012680:Sensitivity and Specificity; D034101:Sporozoites / Q000737:chemistry / Q000254:growth & development / Q000378:metabolism
chemical_list: D015800:Protozoan Proteins; C041595:circumsporozoite protein, Protozoan
keywords: ELISA; Malaria; Oocyst; Plasmodium; Vector screening