Abstract: To solve the problems of complex operation and high cost of traditional PCR methods for early chicken embryos，this study aimed to establish a rapid PCR detection technology based on trace blood samples， so as to provide a new method for optimizing the PCR identification of early poultry embryos.［Methods］Chicken embryos at three incubation time points（6.5，8.5，and 10.5 days）were selected，and primers for sex identification genes，germ cell marker genes，and housekeeping genes were designed for the experiment. Comparative experiments were carried out simultaneously，including traditional tissue DNA extraction，rapid DNA extraction，and direct PCR dete ction with trace blood samples aspirated as templates，and the amplification efficiencies of three types of PCR premixes were compared.［Results］The experimental results showed that：（1）The PCR identification results of trace blood were consistent with those of the traditional tissue DNA extraction method，with an accuracy rate of 100%；（2） The optimized PCR identification system did not need to add PCR enhancers and stabilizers，and whole blood samples could be directly used as templates after dilution，which significantly reduced the detection cost per sample. ［Conclusion］The direct PCR method using trace blood from early chicken embryos established in this study can replace the traditional DNA extraction identification method，which greatly shortens the time and simplifies the operation steps，and significantly improves work efficiency while maintaining 100% accuracy. This technology has the advantages of rapidity，convenience，low cost and reliable results，providing an innovative solution for the PCR identification of poultry embryos.

Key words: Blood PCR；Early chicken embryo；Sex identification；Ultra?fidelity enzyme

摘要： 为解决传统鸡早期胚胎PCR方法操作复杂、成本高的问题，该研究旨在建立基于微 量血液样本的PCR快速检测技术，为优化家禽早期胚胎PCR鉴定提供新方法。【方法】选择3个 孵化时间点（6.5、8.5、10.5 d）的鸡胚并设计性别鉴定基因、生殖细胞标记基因和管家基因的引 物开展试验，同步开展传统组织DNA提取、快速DNA提取和吸取微量血样作为模板直接进行 PCR检测对比实验，并比较3种PCR预混液的扩增效率。【结果】试验结果显示：（1）微量血液与 传统组织DNA提取法的PCR鉴定结果一致，准确率达100%；（2）优化后的PCR鉴定体系无需 添加PCR增强剂和稳定剂，全血样本经稀释即可直接作为模板，单样本检测成本明显降低。 【结论】该研究建立的鸡早期胚胎微量血液直接PCR法可以替代传统DNA提取鉴定方法，极大 地缩短了时间并简化了操作步骤，在保持100%准确率的同时显著提高工作效率。该技术具有 快速便捷、成本低廉、结果可靠等优势，为家禽胚胎PCR鉴定提供了创新解决方案。

关键词: 血液PCR；鸡早期胚胎；性别鉴定；超保真酶