变形杆菌基因分型方法的评价

目的　筛选出适用于变形杆菌的最佳分型方法。方法　用脉冲场凝胶电泳 (PFGE)、随机扩增多态 DNA(RAPD)、重复元件序列 PCR(Rep-PCR)和肠杆菌基因间重复共有序列 PCR(ERIC-PCR) 4种方法对 44株变形杆菌进行基因分型。结果　 RAPD可分为 17型 ,分辨率系数为 0 .93 2 ;Rep -PCR可分为 15型 ,分辨率系数为0 .916;ERIC-PCR可分为 2 3型 ,分辨率系数为 0 .941;PFGE可分为 3 6型 ,分辨率系数为 0 .988。结论　 PFGE、RAPD、Rep-PCR和 ERIC-PCR4种基因分型方法对变形杆菌进行分型 ,其分型力和分辨力均很好 (均 DI>0 .90 ) ,其中 PFGE分辨力最高 ,重复性好 ,但实验费用高 ,时间长 ,对大规模广泛应用有局限性 ;Rep-PCR和 ER-IC-PCR分辨率虽略底于 RAPD,但重复性好 ,结果稳定 ,可广泛应用于多种菌属 ,不需要重复设计引物 ,易于实现标准化。

The Evaluation of Four Genotyping Methods for Proteus spp

OBJECTIVE To find the best genotyping method for Proteus spp. METHODS Forty four isolates of Proteus spp were genotyped by PFGE, RAPD, Rep PCR and ERIC PCR. RESULTS All isolates were typed by RAPD with 17 sub types and the discriminatory power was 0.932; by Rep PCR with 15 and 0.916; by ERIC PCR with 23 and 0.941; by PFGE with 36 and 0.988, respectively. CONCLUSIONS All four genotyping methods of PFGE, RAPD, Rep PCR and ERIC PCR showed high typeability and discriminatory power (DI>0.90 all). PFGE was a method with the highest discriminatory power and reproducibility, but not suitable for large scale screening due to its disadvantages of high cost and time consuming. Rep PCR and ERIC PCR were relatively more useful than PFGE and RAPD for its stable reproducibility, widely use in several bacterium genus, easy to implement, low cost and time saving, although its discriminatory rate was little less than PFGE and RAPD.