A simple and rapid method for fish sex identification based on recombinase-aided amplification and its use in Cynoglossus semilaevis

Abstract Fish sex identification is a basic technique of great importance for both fish genetic studies and fisheries. Due to the sexual reversal phenomenon in many fish species, a simple and rapid molecular identification method for fish genetic sex is urgently needed to suit versatile detection sc...

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Bibliographic Details
Main Authors: Zhichao Nie, Peng Lü, Rusong Zhang, Yishuai Tu, Zhenni Liu, Yin Li, Cong Tang, Xiqing Li, Kun Zhao, Qiang Zhou, Feng Li, Jun Wang, Zhanzhuang Zeng, Min Tu, Hong Zhang
Format: article
Language:EN
Published: Nature Portfolio 2021
Subjects:
R
Q
Online Access:https://doaj.org/article/e2c942828f8543e4a093e6f336676548
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Summary:Abstract Fish sex identification is a basic technique of great importance for both fish genetic studies and fisheries. Due to the sexual reversal phenomenon in many fish species, a simple and rapid molecular identification method for fish genetic sex is urgently needed to suit versatile detection scenarios, such as point-of-need applications. In this study, we took Cynoglossus semilaevis as an example, established a recombinase-aided amplification (RAA)-based method for sex identification, and combined the RAA-detection with two result visualization approaches with distinct features, capillary electrophoresis (CE) and lateral flow dipstick (LFD). Specific primers and probe were designed to specifically detect the sex chromosome W of C. semilaevis in order to distinguish the genetic sex between males, pseudo-males and females. To evaluate the performance of our methods, the genetic sex for twenty-eight males, sixty-eight pseudo-males and fifty-four females were examined with the RAA-based method and classical PCR-based genotyping method, demonstrating the consistent results of sex identification between both methods. The RAA-LFD method is operationally simple, rapid (~ 30 min) and holds great potential for point-of-need applications of fish sex identification, including fishery fields. The method presented here could be effective for identifying fish gender with the ZW karyotype.