Fiber-optic microarray for simultaneous detection of multiple harmful algal bloom species.
Appl Environ Microbiol
; 72(9): 5742-9, 2006 Sep.
Article
en En
| MEDLINE
| ID: mdl-16957189
ABSTRACT
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 mum) 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.
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Fitoplancton
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Análisis de Secuencia por Matrices de Oligonucleótidos
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Eutrofización
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Eucariontes
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
Appl Environ Microbiol
Año:
2006
Tipo del documento:
Article
País de afiliación:
Estados Unidos