Bioluminescence DNA hybridization assay for Plasmodium falciparum based on the photoprotein aequorin

Leslie Doleman, Logan Davies, Laura Rowe, Elizabeth A. Moschou, Sapna K Deo, Sylvia Daunert

Research output: Contribution to journalArticle

32 Citations (Scopus)

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 ∼50 parasites/μL 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/μL 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.

Original languageEnglish
Pages (from-to)4149-4153
Number of pages5
JournalAnalytical Chemistry
Volume79
Issue number11
DOIs
StatePublished - Jun 1 2007
Externally publishedYes

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Luminescent Proteins
Aequorin
Bioluminescence
Assays
DNA
Labels
Throughput
Streptavidin
DNA Probes
Optical microscopy
Amplification
Signal to noise ratio
Blood
Personnel

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Bioluminescence DNA hybridization assay for Plasmodium falciparum based on the photoprotein aequorin. / Doleman, Leslie; Davies, Logan; Rowe, Laura; Moschou, Elizabeth A.; Deo, Sapna K; Daunert, Sylvia.

In: Analytical Chemistry, Vol. 79, No. 11, 01.06.2007, p. 4149-4153.

Research output: Contribution to journalArticle

Doleman, Leslie ; Davies, Logan ; Rowe, Laura ; Moschou, Elizabeth A. ; Deo, Sapna K ; Daunert, Sylvia. / Bioluminescence DNA hybridization assay for Plasmodium falciparum based on the photoprotein aequorin. In: Analytical Chemistry. 2007 ; Vol. 79, No. 11. pp. 4149-4153.
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