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

Yuki Minegishi, Tatsuki Yoshinaga, Jun Aoyama, Katsumi Tsukamoto

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

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 l-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.

Original languageEnglish
Pages (from-to)1915-1918
Number of pages4
JournalICES Journal of Marine Science
Volume66
Issue number9
DOIs
Publication statusPublished - 2009 Oct 1

Keywords

  • Anguilla japonica
  • Cycles at threshold (Ct)
  • Sequence detection system
  • Species identification
  • Template DNA concentrations

ASJC Scopus subject areas

  • Oceanography
  • Ecology, Evolution, Behavior and Systematics
  • Aquatic Science
  • Ecology

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