Development Of Dot Blot Hybridization Method For Identification Of The Brucella Live Vaccine Strain S2

To discriminate the Brucella live vaccine strain S2 from field strains, the dot blot hybridization was explored. According to the difference of geneme between Brucella live vaccine strain S2 and other Brucella strains in NCBI, specific primers and 29 bp probe were designed and labeled by DIG High Prime. The PCR product was recovered, purified, denatured and fixed to the NC membrane at 120℃ in 30 min, then hybridized with the probe labeled by DIG High Prime and detected. The fragment of 330 bp could be amplified from strain S2, and 360 bp from other Brucella strains. DIG could be connected to the oligonucleotides successfully by method of end labeling. The probe was specific and no cross hybridization with other Brucella Strains and could be repeated. 10 pg DNA of Brucella live vaccine strain S2 could be discriminated in 10 h by the dot blot hybridization.