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

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.

基于聚合酶链式反应（Polymerase Chain Reaction，PCR）的方法是当前鸟类性别鉴定最常用的技术手段。尽管此类方法具备快速、简便且准确的优势，但仍需依赖专用实验设施，导致其无法在野外场景中开展应用。由此带来的采样与检测结果获取之间的时间差，会阻碍研究人员结合个体性别信息在野外现场及时做出决策。本研究提出一种基于环介导等温扩增（Loop-mediated isothermal amplification，LAMP）技术的鸟类野外性别鉴定方法，通过扩增性别染色体特异性重复基因Chromo-Helicase-DNA结合蛋白（Chromo-Helicase-DNA binding protein）实现性别鉴定。我们以鹰科（Accipitridae）的白背秃鹫（Gyps fulvus）、埃及秃鹫（Neophron percnopterus）以及黑鸢（Milvus migrans）为对象验证了本方法。本研究首次提出适用于野生动物野外性别鉴定的实地操作流程，使用同一套特异性LAMP引物，成功完成了三个不同猛禽亚科物种的性别鉴定。该分子检测技术可直接部署于采样现场：仅需通过电压逆变器将恒温金属浴模块连接至汽车点烟器即可实现温控，反应管内的颜色变化可通过裸眼直接读取检测结果。引物与试剂可提前配制完成，便于室温储存。我们提供了详细的操作实施指南，相较于基于PCR的实验室检测方法，该流程更为简便（无需电泳操作）、成本更低且耗时更短（检测结果可在约90分钟内获得）。本研究在三个不同猛禽亚科中实现跨物种成功应用，表明这套标记物有望成为其他猛禽科属分子性别鉴定的实用工具，同时该野外操作流程可推广至所有鸟类物种。