关键词: 鹅；重组聚合酶扩增；性别鉴定；快速检测

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