鸭疫里氏杆菌鸭源致病性大肠埃希氏菌和鸭沙门菌的RAPD研究

以 4株不同血清型的鸭疫里氏杆菌、5株不同血清型的鸭源致病性大肠埃希氏菌和 5株同一血清型的鸭沙门菌为研究对象 ,分别提取基因组DNA ,利用随机扩增多态性DNA(RAPD)技术对其基因组DNA进行了分析。结果 ,从 2 0条随机引物中筛选出了 38条能在这 3种菌中有较好多态性的随机引物 ,8个有效引物共扩增出了 10 2个DNA片段 ,其中 14个菌株共有的谱带仅有 1条 ,显示多态性的片段有 10 1个 ,占 99.0 % ;对RA的 4个菌株扩增出的谱带数为 5 1条 ,共同片段有 12条 ,多态性片段 39条 ,占 76 .5 % ;对沙门菌的 5个菌株扩增出的条带数为 4 6条 ,共同片段为13条 ,多态性片段 33条 ,占 71.7% ;对大肠埃希氏菌的 5个菌株扩增出的条带数为 4 8条 ,共同片段 4条 ,多态性片段 4 4条 ,占 91.7%。在 8条引物中进一步筛选出 1条引物G12 ,其图谱中 5 35bp的条带为鸭疫里氏杆菌所特有 ,330bp的条带为沙门菌所特有 ,12 2 7bp的条带为大肠埃希氏菌所特有 ,表明G12可作为分子标记用来鉴别这 3种细菌。

Study on Riemerella anatipestifer,enteropathogenic Escherichia coli and Salmonella anatum from ducks by random amplified polymorphic DNA

The random amplified polymorphic DNA (RAPD) was used to study genetic diversity (among) four serotype Riemerella anatipestifer strains, five enteropathogenic Escherichia coli strains and five Salmonella anatum strains from ducks. 8 primers were selected from twenty 10 bp random primers. Totally 102 RAPD bands were identified, 1 of which was common all the accessions and 101 of which were polymorphic between any two accessions. The polymorphic bands accounted for 99.0% of the total amplified bands. 51 bands were from 4 strains of R.anatipestifer , 12 of which were common, and polymorphic rate was 76.5%. 46 bands were from 5 strains of S.anatum , 13 of which were common, and polymorphic rate was 71.7%. 48 bands were from 4 strains of E.coli, 4 of which were common, and polymorphic rate was 91.7%. The 535 bp band, the 1 227 bp band and the 330 bp band, which were amplified with the primer G12 selected from the above-mentioned eight primers, were characteristic for R.anatipestifer, E.coli and S.anatum respectively. The results showed that the primer G12 can be used as molecular marker to differentiate R.anatipestifer from E.coli or S.anatum and to differentiate E.coli from S.anatum.