title: Bioluminescence DNA hybridization assay for Plasmodium falciparum based on the photoprotein aequorin.
abstract: A bioluminescence DNA hybridization assay for the detection of Plasmodium falciparum, the most deadly species of malaria, using the photoprotein aequorin as a bioluminescent label has been developed. The current gold standard for the detection of malaria is light microscopy, which can detect down to approximately 50 parasites/microL of blood, but has low-throughput, high costs, and requires high skill, which limit the applicability of the method, especially in the developing regions where malaria detection is mostly needed. The utilization of aequorin as a bioluminescence label offers the advantages of high signal-to-noise ratio and reliable detection down to attomole levels, allowing for the development of highly sensitive and miniaturized high-throughput bioluminescence assays. Herein, we developed a DNA hybridization assay for the detection of P. falciparum based on the competition between the target DNA and the signal generating DNA streptavidin-aequorin for hybridization with the probe DNA. This bioluminescence hybridization assay demonstrated a detection limit of 3 pg/microL and was employed for the detection of target DNA in standard and spiked human serum samples. The DNA hybridization assay was developed in a microplate format without the need for sample PCR amplification, showing the potential suitability of this method in the parallel analysis of samples by low-trained personnel, such as that typically encountered in developing regions.
authors.affiliation: Department of Chemistry, University of Kentucky, Lexington, Kentucky 40506-0055, USA.
authors.forename: Leslie
authors.initials: L
authors.lastname: Doleman
authors.forename: Logan
authors.initials: L
authors.lastname: Davies
authors.forename: Laura
authors.initials: L
authors.lastname: Rowe
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Moschou
authors.forename: Sapna
authors.initials: S
authors.lastname: Deo
authors.forename: Sylvia
authors.initials: S
authors.lastname: Daunert
mesh_terms: D000331:Aequorin / Q000032:analysis* / Q000235:genetics*; D000818:Animals; D015342:DNA Probes / Q000032:analysis* / Q000235:genetics*; D016054:DNA, Protozoan / Q000032:analysis* / Q000235:genetics*; D008163:Luminescent Measurements / Q000379:methods*; D009690:Nucleic Acid Conformation; D010963:Plasmodium falciparum
chemical_list: D015342:DNA Probes; D016054:DNA, Protozoan; D000331:Aequorin