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ICES Journal of Marine Science: Journal du Conseil Advance Access originally published online on June 4, 2009
ICES Journal of Marine Science: Journal du Conseil 2009 66(9):1915-1918; doi:10.1093/icesjms/fsp158
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© 2009 International Council for the Exploration of the Sea. Published by Oxford Journals. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Species identification of Anguilla japonica by real-time PCR based on a sequence detection system: a practical application to eggs and larvae

Yuki Minegishi1, Tatsuki Yoshinaga2, Jun Aoyama1 and Katsumi Tsukamoto1

1 Ocean Research Institute, University of Tokyo, 1-15-1 Minamidai, Nakano-ku, Tokyo 164-8639, Japan
2 School of Marine Biosciences, Kitasato University, Ofunato, Iwate 022-0101, Japan

Correspondence to Y. Minegishi: tel: +81 3 5351 6512; fax: +81 3 5351 6514; e-mail: minegisi{at}ori.u-tokyo.ac.jp

Minegishi, Y., Yoshinaga, T., Aoyama, J., and Tsukamoto, K. 2009. Species identification of Anguilla japonica by real-time PCR based on a sequence detection system: a practical application to eggs and larvae. – ICES Journal of Marine Science, 66: 1915–1918.

To develop a practical method for identifying Japanese eel Anguilla japonica eggs and larvae to species by a sequence detection system using a real-time polymerase chain reaction (PCR), we examined (i) the sensitivity of the system using samples at various developmental stages, and (ii) influences of intra- and interspecific DNA sequence variations in the PCR target region. PCR amplifications with extracted DNA solution at 7.0 ng µl–1 or lower were efficient at distinguishing A. japonica from other anguillids. A single egg at the gastrula or later developmental stages could also be identified. Two sequence variations in the PCR target region were observed in 2 out of 35 A. japonica collected from three localities, and from four year classes at a single locality. These mutations, however, did not affect the result of species identification achieved by A. japonica-specific PCR primers and probe. The accuracy of this PCR-based method of species identification will help in field surveys of the species.

Keywords: Anguilla japonica, cycles at threshold (Ct), sequence detection system, species identification, template DNA concentrations

Received 2 March 2009; accepted 30 April 2009; advance access publication 4 June 2009.


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