中国实验动物学报Issue(3)：72-77,6.DOI:10.3969/j.issn.1005-4847.2014.03.015
反向斑点杂交检测泰泽病原体
Application of reverse dot blotting for detection of Tyzzer’ s organism
摘要
Abstract
Objective To establish a simple, stable, specific and sensitive method for detection of Tyzzer ’ s or-ganism by reverse dot blotting ( RDB) .Methods Primers and specific probes were designed according to the conservative sequence of Tyzzer 16S rDNA.The forward primer was labeled with biotin .The reverse dot blotting method was established followed by PCR amplification .The specificity and sensitivity of this method were determined .Next, 41 mice and 38 rats were examined by RDB , ELISA and IFA .Results The RDB method showed a high specificity , and in the testing of the 79 laboratory animals , its limit of detection was 4.5 ng/μL.Compared the results of ELISA and IFA , its consistence with ELISA was 100%and the positive rate was 7.59%(6/79), the consistence with IFA was 92.4%(73/79), and the posi-tive rate was 0%.Conclusions An accurate, sensitive and specific method in combination with PCR and RDB in detection of Tyzzer’ s organism is established in this study .关键词
泰泽病原体/反向斑点杂交/实验动物Key words
Tyzzer’s organism/Reverse dot blot/Laboratory animals分类
生物科学引用本文复制引用
赵婷婷,严磊,魏立雯,韦莉,王会娟,赖国旗,谭毅..反向斑点杂交检测泰泽病原体[J].中国实验动物学报,2014,(3):72-77,6.基金项目
重庆市科委科技创新能力建设项目《重庆市啮齿类实验动物工程技术研究中心》CSTC，2010CB5013。 （）
