A no film slot blot for the detection of developing P. Falciparum oocysts in mosquitoes

Bryan Grabias, Nitin Verma, Hong Zheng, Abhai Tripathi, Godfree Mlambo, Merribeth J. Morin, Emily Locke, Sanjai Kumar

Research output: Contribution to journalArticle

Abstract

Non-microscopy-based assays for sensitive and rapid detection of Plasmodium infection in mosquitoes are needed to allow rapid and high throughput measurement of transmission intensity and malaria control program effectiveness. Here, we report on a modified enhanced chemiluminescence-based slot blot assay for detection of Plasmodium falciparum (Pf) circumsporozite protein (PfCSP) expressed on parasite oocysts developing inside the mosquito midgut. This modified assay has several novel features that include eliminating the need for exposure to autoradiography (AR) film, as well as utilizing a novel high affinity anti-CSP antibody, and optimizing assay procedures resulting in significant reduction in the time required to perform the assay. The chemiluminescent signal for the detection of PfCSP in mosquito samples was captured digitally utilizing the C-Digit blot scanner that, allowed the detection of 0.01 pg of recombinant P. falciparum CSP and as few as 0.02 P. falciparum oocysts in a little over two hours. The earlier ECL-SB detected rCSP and oocysts and took approximately 5 h to perform. Whole mosquito lysates from both high and low prevalence infected mosquito populations were prepared and evaluated for PfCSP detection on the ECL-SB by both AR film and digital data capture and analysis. There was a 100% agreement between the AR film and the C-Digit scanner methods for PfCSP detection in randomly sampled mosquitoes. This novel No Film Slot Blot assay obviates the need for AR film exposure and development and significantly reduces the assay time enabling widespread use in field settings.

Original languageEnglish (US)
Article number0174229
JournalPLoS One
Volume12
Issue number4
DOIs
StatePublished - Apr 1 2017

Fingerprint

Oocysts
Culicidae
oocysts
Assays
Autoradiography
autoradiography
assays
Plasmodium falciparum
Emitter coupled logic circuits
scanners
Malaria
Proteins
Malaria control
proteins
Program Evaluation
Luminescence
Chemiluminescence
chemiluminescence
Plasmodium
Anti-Idiotypic Antibodies

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

A no film slot blot for the detection of developing P. Falciparum oocysts in mosquitoes. / Grabias, Bryan; Verma, Nitin; Zheng, Hong; Tripathi, Abhai; Mlambo, Godfree; Morin, Merribeth J.; Locke, Emily; Kumar, Sanjai.

In: PLoS One, Vol. 12, No. 4, 0174229, 01.04.2017.

Research output: Contribution to journalArticle

Grabias, Bryan ; Verma, Nitin ; Zheng, Hong ; Tripathi, Abhai ; Mlambo, Godfree ; Morin, Merribeth J. ; Locke, Emily ; Kumar, Sanjai. / A no film slot blot for the detection of developing P. Falciparum oocysts in mosquitoes. In: PLoS One. 2017 ; Vol. 12, No. 4.
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