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Ko YT, Chan MM, Ford SE, et al. A PCR-ELISA Method for Direct Detection of the Oyster Pathogen Haplosporidium nelsoni. Mar Biotechnol (NY). 1999;1(2):147-154doi: 10.1007/pl00011762.
Ko, Y. T., Chan, M. M., Ford, S. E., & Fong, D. (1999). A PCR-ELISA Method for Direct Detection of the Oyster Pathogen Haplosporidium nelsoni. Marine biotechnology (New York, N.Y.), 1(2), 147-154. https://doi.org/10.1007/pl00011762
Ko, et al. "A PCR-ELISA Method for Direct Detection of the Oyster Pathogen Haplosporidium nelsoni." Marine biotechnology (New York, N.Y.) vol. 1,2 (1999): 147-154. doi: https://doi.org/10.1007/pl00011762
Ko YT, Chan MM, Ford SE, Fong D. A PCR-ELISA Method for Direct Detection of the Oyster Pathogen Haplosporidium nelsoni. Mar Biotechnol (NY). 1999 Mar;1(2):147-154. doi: 10.1007/pl00011762. PMID: 10373623.
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Mar Biotechnol (NY). 1999 Mar;1(2):147-154. doi: 10.1007/pl00011762.

A PCR-ELISA Method for Direct Detection of the Oyster Pathogen Haplosporidium nelsoni.

Marine biotechnology (New York, N.Y.)

Ko, Chan, Ford, Fong

Affiliations

  1. Department of Nutrition, China Medical College, Taichung, Taiwan 40421

PMID: 10373623 DOI: 10.1007/pl00011762

Abstract

: A rapid method, utilizing both polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA), was developed for detection of oyster MSX disease. The technique included using Haplosporidium nelsoni pathogen-specific PCR primers (based on ribosomal RNA genes), a Chelex resin (for rapid DNA extraction from oyster mantle tissues), and cloned H. nelsoni rRNA plasmid DNA (for use as a capture probe). Digoxigenin was incorporated into the pathogen-specific PCR products, which were captured by the coated probe in a fast hybridization reaction and then detected by ELISA. The sensitivity of PCR amplification on cloned plasmid DNA was 10 fg for detection by stained agarose gel, and increased to 0.01 fg for ELISA. Positive signals were observed in infected oysters using the PCR-ELISA technique. This method may be applicable to early detection of infection.

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