Sex determination in the wild: a field application of loop-mediated isothermal amplification successfully determines sex across three raptor species

A. Centeno-Cuadros*, I. Abbasi, R. Nathan

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

22 Scopus citations


PCR-based methods are the most common technique for sex determination of birds. Although these methods are fast, easy and accurate, they still require special facilities that preclude their application outdoors. Consequently, there is a time lag between sampling and obtaining results that impedes researchers to take decisions in situ and in real time considering individuals’ sex. We present an outdoor technique for sex determination of birds based on the amplification of the duplicated sex-chromosome-specific gene Chromo-Helicase-DNA binding protein using a loop-mediated isothermal amplification (LAMP). We tested our method on Griffon Vulture (Gyps fulvus), Egyptian Vulture (Neophron percnopterus) and Black Kite (Milvus migrans) (family Accipitridae). We introduce the first fieldwork procedure for sex determination of animals in the wild, successfully applied to raptor species of three different subfamilies using the same specific LAMP primers. This molecular technique can be deployed directly in sampling areas because it only needs a voltage inverter to adapt a thermo-block to a car lighter and results can be obtained by the unaided eye based on colour change within the reaction tubes. Primers and reagents are prepared in advance to facilitate their storage at room temperature. We provide detailed guidelines how to implement this procedure, which is simpler (no electrophoresis required), cheaper and faster (results in c. 90 min) than PCR-based laboratory methods. Our successful cross-species application across three different raptor subfamilies posits our set of markers as a promising tool for molecular sexing of other raptor families and our field protocol extensible to all bird species.

Original languageAmerican English
Pages (from-to)153-160
Number of pages8
JournalMolecular Ecology Resources
Issue number2
StatePublished - 1 Mar 2017

Bibliographical note

Funding Information:
We are grateful to Mónica Gutiérrez and Ana Píriz for their help, laboratory assistance and expertise during the experimental design and laboratory work as well as to Martina Carrete, José Luis Tella, Miguel Delibes and Pim Edelaar for their support and encouragement. We also thank Orr Spiegel, Roi Harel, Ohad Hatzofe, Lidia López and Julio Blas for providing samples and all members of the Movement Ecology Lab for their helpful comments. Samples of Griffon and Egyptian vultures were provided by José A. Donázar as principal investigator of projects RNM 1925 and CGL2012-40013-C02-01, respectively. This work was funded by the Severo Ochoa Award, the U.S.–Israel Bi-national Science Foundation (BSF 255/2008), the special BSF Multiplier Grant Award from the Rosalinde and Arthur Gilbert Foundation, the Adelina and Massimo Della Pergola Chair of Life Sciences and the Minerva Center for Movement Ecology. Logistical support was provided by Laboratorio de Ecología Molecular, Estación Biológica de Doñana, CSIC (LEM-EBD). A.C-C. was granted by the Lady Davis Fellowship Trust and the University Pablo de Olavide.

Publisher Copyright:
© 2016 John Wiley & Sons Ltd


  • CHD-W
  • CHD-Z
  • Gyps fulvus
  • Milvus migrans
  • Neophron percnopterus
  • loop-mediated isothermal amplification


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