Abstract: This study aimed to establish a rapid on?site method for sex identification of goslings to address the increased feeding costs caused by mixed?sex rearing in goose production，as well as the limitations of traditional sexing methods，including time consumption，high cost，and low accuracy. Recombinase polymerase amplification（RPA）is an emerging isothermal nucleic acid amplification technology. Compared with conventional amplification methods， RPA offers advantages such as high sensitivity，high amplification efficiency，and simple operational requirements. The HINTW gene was selected as the amplification target，and specific RPA primers were designed to amplify DNA extracted from the blood of male and female Lion?head geese. Additionally，a rapid DNA extraction method was explored in which whole blood，heated at 100 ℃ for 5 minutes，was used as a template for the RPA reaction as a substitute for kit?extracted DNA. The results showed that the designed primers exhibited high specificity for female geese. Agarose gel electrophoresis revealed a single band for all female individuals，whereas no band was observed for any male individual. The sensitivity of the RPA assay，as determined by agarose gel electrophoresis，reached 4.7×10-1 ng/μL. The results of RPA detection using heat?treated whole blood as a template were consistent with those obtained using extracted DNA as a template. This study demonstrates that the method can achieve rapid amplification of the female?specific HINTW gene under a constant temperature of 39 ℃ with simple amplification conditions，making it a viable on?site rapid sex identification method for goslings.

Key words: Goose；Recombinase Polymerase Amplification；Sex Identification；Rapid Detection

摘要： 研究旨在建立一种雏鹅性别现场快速检测方法，以解决鹅类养殖生产中雌雄混养 导致的饲养成本增加，传统性别检测方法中存在耗时长、成本高、准确率低等问题。重组聚合 酶扩增技术（Recombinase polymerase amplification，RPA）是一种新兴的核酸恒温扩增技术，和 传统的扩增技术相比，该技术具有灵敏度高、扩增效率高以及操作条件简单等优势，研究选取 HINTW基因作为扩增目标，设计了RPA特异性引物，对雌雄狮头鹅血液提取的DNA进行RPA 扩增；另外研究还探索了DNA的快速提取方法，通过100 ℃的恒温环境下加热5 min处理的全 血代替试剂盒提取的DNA进行RPA反应。结果显示：该引物对雌鹅具有高度特异性，琼脂糖 凝胶电泳结果提示，雌鹅个体均显示单一条带，雄鹅个体均无条带；琼脂糖凝胶电泳检测的 RPA反应灵敏度可达4.7×10-1 ng/μL；利用加热法处理的全血作为模板进行的RPA检测结果与 利用DNA作为模板产生的结果一致。研究表明：该方法可以在39 ℃的恒温下完成对雌鹅特异 性基因HINTW的快速扩增，且扩增所需条件简单，可以作为雏鹅性别现场快速检测方法之一。

关键词: 鹅；重组聚合酶扩增；性别鉴定；快速检测