Publication Date:
2022-05-25
Description:
Author Posting. © American Society for Microbiology, 2006. This article is posted here by permission of American Society for Microbiology for personal use, not for redistribution. The definitive version was published in Applied and Environmental Microbiology 72 (2006): 5742-5749, doi:10.1128/AEM.00332-06.
Description:
Harmful algal blooms (HABs) are a serious threat to coastal resources, causing a variety of impacts on public health, regional economies, and ecosystems. Plankton analysis is a valuable component of many HAB monitoring and research programs, but the diversity of plankton poses a problem in discriminating toxic from nontoxic species using conventional detection methods. Here we describe a sensitive and specific sandwich
hybridization assay that combines fiber-optic microarrays with oligonucleotide probes to detect and enumerate the HAB species Alexandrium fundyense, Alexandrium ostenfeldii, and Pseudo-nitzschia australis. Microarrays were prepared by loading oligonucleotide probe-coupled microspheres (diameter, 3 μm) onto the distal ends
of chemically etched imaging fiber bundles. Hybridization of target rRNA from HAB cells to immobilized
probes on the microspheres was visualized using Cy3-labeled secondary probes in a sandwich-type assay
format. We applied these microarrays to the detection and enumeration of HAB cells in both cultured and field
samples. Our study demonstrated a detection limit of approximately 5 cells for all three target organisms
within 45 min, without a separate amplification step, in both sample types. We also developed a multiplexed
microarray to detect the three HAB species simultaneously, which successfully detected the target organisms,
alone and in combination, without cross-reactivity. Our study suggests that fiber-optic microarrays can be used
for rapid and sensitive detection and potential enumeration of HAB species in the environment.
Description:
This work was funded by the Sea Grant Technology Program (NA16RG2273).
Repository Name:
Woods Hole Open Access Server
Type:
Article
Format:
328437 bytes
Format:
application/pdf
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