Application of 16S rDNA Real-time Fluorescent Quantitate PCR for Detecting Gamma Germ

作     者：HUANG Yongcheng,CHENG Anchun,WANG Mingshu,YANG Xiaoyan,QI Xuefeng, GE Zhongyuan,GUO Yufei,ZHANG Suhui,HUji,CHEN Xiaoyue (Avian Disease Research Center, College of Animal Science and Veterinary Medicine of Sichuan Agricultural University Key Laboratory of Animal Disease and Human Health of Sichuan Province, Yaan, Sichuan, China,625014) 

会议名称：《第三届第八次全国学术研讨会暨动物微生态企业发展战略论坛》

会议日期：2006年

学科分类：090603[农学-临床兽医学] 09[农学] 0906[农学-兽医学] 

关 键 词：Bacillus 16S rDNA Real time fluorescent quantitate PCR 

摘      要：Group-specificity quantitate PCR primers and probe were designed and synthesized ,and method of detecting Bacillus was *** optimal Mg2+ concentration of the real-time fluorescent quantitate PCR was 7.5 mmol/L, and the optimal probe concertration was 0.1 μmol/L;The detecting of Bacillus pumilus with the method was masculine, but to Bifidobacterium bifidus,***, Lactobacillus bulgaricus, Enterococcus faecium,and Staphylococcus aureus those were existed in animal intestinal tract was negative;the lowest detect examine was 8 copies of 16S rDNA;Extracted DNA of the duodenal, jejunal, ileac, cecal ,and rectal contents of ducks at 20,21, 22,23,26,32,41 ,and 56d of age was detected with this method , results indicated these intestinal tract spots has the bacillus to exist, the quantity of Bacillus in different intestinal tract spot has not obvious difference. This method has high sensitivity and specificity, which could detect fast the quantity of Bacillus in the duck intestinal tract.