Application of CRISPR-Based Diagnostics for Malaria Antigen Detection: Feasibility Assessment in a Nigerian Context

CRISPR-based diagnostic platforms, including SHERLOCK (Specific High-sensitivity Enzymatic Reporter unLOCKing) and DETECTR (DNA Endonuclease Targeted CRISPR Trans Reporter), represent next-generation nucleic acid diagnostic technologies with potential for application in malaria-endemic low-resource settings. This study assessed the technical feasibility of applying a CRISPR-Cas13a-based assay for Plasmodium falciparum detection in clinical blood samples from febrile patients attending hospitals in Abeokuta, Ogun State, Southwest Nigeria. One hundred blood samples were collected and tested in parallel with the CRISPR-Cas13a assay targeting P. falciparum 18S rRNA and a reference nested PCR method. Rapid diagnostic test (RDT) results were also recorded for comparison. CRISPR-Cas13a demonstrated sensitivity of 96.2% and specificity of 97.6% compared to nested PCR. The assay detected infections at parasite densities below the RDT detection threshold in 11% of PCR-confirmed cases. Sample-to-result time was 75 minutes, with a simple visual fluorescence output compatible with battery-powered portable illuminators. Cold chain stability of lyophilized CRISPR reagents over 72 hours at ambient temperature was maintained. This feasibility study supports the potential integration of CRISPR diagnostics into malaria surveillance programs in Southwest Nigeria, pending cost-reduction and regulatory pathway development. Keywords: CRISPR diagnostics, SHERLOCK, Plasmodium falciparum, malaria detection, Ogun State.