带毒蚕卵中的家蚕微孢子虫DNA提取方法研究  

The Method for Extraction of Nosema bombycis DNA From Silkworm Eggs With Pebrine

作　　者：潘中华[1] 郑小坚[1] 薛仁宇[1] 曹广力[1] 贡成良[1] 陶维华[2] 李掌林[2] 陶鸣[2] 潘丽芬[2] 

机构地区：[1]苏州大学生命科学学院,苏州215123 [2]江苏省蚕种管理所,无锡214151

出　　处：《蚕业科学》2005年第4期486-489,共4页ACTA SERICOLOGICA SINICA

基　　金：国家茧丝办风险基金[编号国茧丝办(2002)36号]

摘　　要：为促进用PCR法检测家蚕微粒子病的生产应用，对带毒蚕卵中的家蚕微孢子虫DNA提取方法进行了研究。结果表明，蚕卵经30％KOH、27℃预处理后抽提的核酸能满足PCR检测对DNA质量的要求。以该方法抽提的DNA为模板，进行蚕卵集团检测，结果10—100粒蚕卵中混有1粒“有毒”卵，即能用PCR的方法检测出。“1／100”“有毒”卵的检出概率约为80％。The method for extracting Nosema bombycis DNA from Bombyx mori eggs infected with Nosema bombycis was studied to approach the feasible application of detecting through PCR. The results showed that the nucleic acid extracted from Bombyx mori eggs with 30% KOH at 37 ℃ pretreatment can match the needs of pebrine detection by PCR. The DNA extracted with this method was used as template to carry out pebrine detection from mass eggs; one infected egg mixed into 10 to 100 pebrine-free eggs can be detected by PCR. The positive probability of pebrine detection is about 80% when one infected egg mixed with 100 normal eggs.

关 键 词：家蚕微粒子病 蚕卵 DNA抽提 PCR检测 

分 类 号：S884.2[农业科学—特种经济动物饲养]