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FANG Qin-mei, ZHU Ji-chang, CHI Hong-shu, LIN Tian-long. Establishment and Optimization of PCR-ELISA Detection Method for Streptococcus Suis Type 2[J]. Fujian Journal of Agricultural Sciences, 2012, 27(12): 1275-1279. doi: 10.19303/j.issn.1008-0384.2012.12.001
Citation:
FANG Qin-mei, ZHU Ji-chang, CHI Hong-shu, LIN Tian-long. Establishment and Optimization of PCR-ELISA Detection Method for Streptococcus Suis Type 2[J]. Fujian Journal of Agricultural Sciences, 2012, 27(12): 1275-1279. doi: 10.19303/j.issn.1008-0384.2012.12.001
FANG Qin-mei, ZHU Ji-chang, CHI Hong-shu, LIN Tian-long. Establishment and Optimization of PCR-ELISA Detection Method for Streptococcus Suis Type 2[J]. Fujian Journal of Agricultural Sciences, 2012, 27(12): 1275-1279. doi: 10.19303/j.issn.1008-0384.2012.12.001
Citation:
FANG Qin-mei, ZHU Ji-chang, CHI Hong-shu, LIN Tian-long. Establishment and Optimization of PCR-ELISA Detection Method for Streptococcus Suis Type 2[J]. Fujian Journal of Agricultural Sciences, 2012, 27(12): 1275-1279. doi: 10.19303/j.issn.1008-0384.2012.12.001
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.
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