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 94℃,denaturation time of 1 min,probe concentration 0.5 μmol·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.