Food Analytical Methods

, Volume 9, Issue 7, pp 2036–2041 | Cite as

Probe-Based Fluorescence Melting Curve Analysis for Differentiating Larimichthys polyactis and Larimichthys crocea

  • Jeong Jin Ahn
  • Youngjoo Kim
  • Ji Young Hong
  • Gi Won Kim
  • Seol Young Kim
  • Seung Yong HwangEmail author


Larimichthys polyactis (redlip yellow croaker) and Larimichthys crocea (large yellow croaker) are commercially important fish species in East Asia with high differences in their market values. In Korea, consumers prefer L. polyactis to L. crocea, although it is difficult to distinguish them based on their morphological traits. The objective of this study was to develop an assay for differentiating L. polyactis and L. crocea using fluorescence melting curve analysis (FMCA) with a single locked nucleic acid (LNA) probe. Species-specific regions of the mitochondrial 16S rDNA were selected as LNA probes. The target sequences of L. polyactis and L. crocea had a 2-bp difference, and a single LNA probe was identified using melting temperature (Tm) shift. LNA probe was 100 % complementary to the target sequence of ten L. polyactis samples, giving a significantly higher Tm value (66 °C) than that of five L. crocea samples (42 °C). Overall, the developed LNA-based FMCA system had high efficiency, multiplexity, and simplicity, and could be effectively used for differentiating L. polyactis and L. crocea, and as a food analyzing method based on DNA sequence.


Larimichthys crocea Larimichthys polyactis Croaker Species identification Fluorescence melting curve analysis Locked nucleic acid 



This study was supported by the National Research Foundation of Korea (NRF), Ministry of Education, Science and Technology, Republic of Korea (No. 2012R1A2A2A03045008)

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Research Involving Human Participants and/or Animals

This article does not contain any studies with human participants performed by any of the authors.

Informed Consent

This article does not contain any studies with human participants performed by any of the authors.


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Jeong Jin Ahn
    • 1
  • Youngjoo Kim
    • 1
  • Ji Young Hong
    • 1
  • Gi Won Kim
    • 2
  • Seol Young Kim
    • 1
  • Seung Yong Hwang
    • 1
    • 2
    Email author
  1. 1.Department of Bio-NanotechnologyHanyang UniversityAnsanKorea
  2. 2.Department of Molecular and Life ScienceHanyang UniversityAnsanKorea

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