福建农业学报2012，Vol.27Issue(12)：1275-1279,5.
猪链球菌2型PCR-ELISA检测方法的建立与优化
Establishment and Optimization of PCR-ELISA Detection Method for Streptococcus Suis Type 2
摘要
Abstract
A new PCR-ELISA detection method of Streptococcus suis type 2 has been set up. A pair of primers contained upstream biotin label and a digoxin-marked specific probe were designed according to the published Streptococcus suis type 2 cps2j gene sequences, PCR amplifier was used as the hybrid spaces, and several main parameters were optimized. The results showed that the concentration of salmon sperm DNA of 1. 6mg/mL, denaturation temperature of 94C , denaturation time of 1 min, probe concentration 0. 5 μmol o mL-1 , hybridization temperature of 55℃ , the the hybridization time 1 min, the first anti-role time 45 min, the second anti-role 40 min were the best conditions for sensitivity and specificity. 17 negative samples were measured whose average was 0. 057, variance was 0. 047, and the ritical value was 0. 198.关键词
猪链球菌2型、PCR-ELISA/cps2j基因Key words
Streptococcus suis type 2/PCR-ELISA/ cps2j gene分类
农业科技引用本文复制引用
方勤美,朱继昌,池洪树,林天龙..猪链球菌2型PCR-ELISA检测方法的建立与优化[J].福建农业学报,2012,27(12):1275-1279,5.基金项目
福建省科技计划项目(2006NZ0003-2、2006F3024) （2006NZ0003-2、2006F3024）
