Bioluminescence DNA Hybridization Assay for Plasmodium falciparum Based on the Photoprotein Aequorin
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- 作者：Leslie Doleman ; Logan Davies ; Laura Rowe ; Elizabeth A. Moschou ; Sapna Deo ; Sylvia Daunert
- 刊名：Analytical Chemistry
- 出版年：2007
- 出版时间：June 1, 2007
- 年：2007
- 卷：79
- 期：11
- 页码：4149 - 4153
- 全文大小：189K
- 年卷期：v.79,no.11(June 1, 2007)
- ISSN：1520-6882
文摘
A bioluminescence DNA hybridization assay for the detection of Plasmodium falciparum, the most deadly species of malaria, using the photoprotein aequorin as abioluminescent label has been developed. The currentgold standard for the detection of malaria is light microscopy, which can detect down to ~50 parasites/
L ofblood, but has low-throughput, high costs, and requireshigh skill, which limit the applicability of the method,especially in the developing regions where malaria detection is mostly needed. The utilization of aequorin as abioluminescence label offers the advantages of high signal-to-noise ratio and reliable detection down to attomolelevels, allowing for the development of highly sensitive andminiaturized high-throughput bioluminescence assays.Herein, we developed a DNA hybridization assay for thedetection of P. falciparum based on the competitionbetween the target DNA and the signal generating DNAstreptavidin-aequorin for hybridization with the probeDNA. This bioluminescence hybridization assay demonstrated a detection limit of 3 pg/L and was employed forthe detection of target DNA in standard and spiked humanserum samples. The DNA hybridization assay was developed in a microplate format without the need for samplePCR amplification, showing the potential suitability of thismethod in the parallel analysis of samples by low-trainedpersonnel, such as that typically encountered in developing regions.
L ofblood, but has low-throughput, high costs, and requireshigh skill, which limit the applicability of the method,especially in the developing regions where malaria detection is mostly needed. The utilization of aequorin as abioluminescence label offers the advantages of high signal-to-noise ratio and reliable detection down to attomolelevels, allowing for the development of highly sensitive andminiaturized high-throughput bioluminescence assays.Herein, we developed a DNA hybridization assay for thedetection of P. falciparum based on the competitionbetween the target DNA and the signal generating DNAstreptavidin-aequorin for hybridization with the probeDNA. This bioluminescence hybridization assay demonstrated a detection limit of 3 pg/L and was employed forthe detection of target DNA in standard and spiked humanserum samples. The DNA hybridization assay was developed in a microplate format without the need for samplePCR amplification, showing the potential suitability of thismethod in the parallel analysis of samples by low-trainedpersonnel, such as that typically encountered in developing regions.