家蚕微孢子DNA的PCR检测灵敏度研究

Study on the Detection Sensitivity of PCR Method for Silkworm Nosema Bombycis DNA

对纯化的家蚕微孢子的DNA抽提及PCR检测灵敏度进行了研究。结果显示:以0.1mol/L K2CO3+0.1mol/L KHCO3作为诱导剂诱导人工纯化的微孢子发芽是最佳选择,配合proteaseK法,能较好地抽提微孢子的DNA;PCR检测灵敏度研究发现,最低检测微孢子核酸浓度是3.25×10-2pg(25μl反应体系),溶液中只要有4粒微孢子即可检出。

This paper has made a study of the method from pure Nosema bombycis abstracting DNA and detection sensitivity of PCR for its DNA. The research result shows that it is the best choice to abstract the DNA from pure Nosema bombycis by using the 0.1 mol/L K2C03 ＋ 0.1 mol/L KHCO3 as the inducement. And matching with the protease K method,we can abstract the Nosema bombycis DNA well. A research has been conducted on the detection sensitivity of PCR method for silkworm Nosema bombycis DNA. Its result shows that： detecting limit is 3.25 ×10^-2 pg（25μl） , 4 grain Nosema bombycis.