Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Old Soutpan Road, Onderstepoort, 0110, Pretoria, South Africa.
Vet Microbiol. 2010 Sep 28;145(1-2):172-6. doi: 10.1016/j.vetmic.2010.02.036. Epub 2010 Mar 7.
Electrophoretic techniques that can be used for genotyping of bacterial pathogens ranges from manual, low-cost, agarose gels to high-throughput capillary electrophoresis sequencing machines. These two methods are currently employed in the electrophoresis of PCR products used in multiple locus VNTR (variable number of tandem repeats) analysis (MLVA), i.e. the agarose electrophoresis (AE) and the capillary electrophoresis (CE). Some authors have suggested that clusters generated by AE are less reliable than those generated by CE and that the latter is a more sensitive technique than the former when typing Mycobacterium tuberculosis complex (MTC) isolates. Because such a claim could have significant consequences for investigators in this field, a comparison was made on 19 Belgian Mycobacterium bovis strains which had previously been genotyped using CE VNTR analysis. The VNTR profiles of the CE VNTR analysis were compared with those obtained by AE VNTR analysis at 14 VNTR loci. Our results indicated that there were no differences in copy numbers at all loci tested when the copy numbers obtained by the AE VNTR analysis were compared with those obtained by CE VNTR analysis. The use of AE VNTR analysis in mycobacterial genotyping does not alter the sensitivity of the MLVA technique compared with the CE VNTR analysis. The AE VNTR can therefore be regarded as a viable alternative in moderately equipped laboratories that cannot afford the expensive equipment required for CE VNTR analysis and data obtained by AE VNTR analysis can be shared between laboratories which use the CE VNTR method.
可用于细菌病原体基因分型的电泳技术包括从手动、低成本的琼脂糖凝胶到高通量毛细管电泳测序仪。这两种方法目前都用于多重位点 VNTR（可变串联重复数）分析（MLVA）的 PCR 产物的电泳，即琼脂糖电泳（AE）和毛细管电泳（CE）。一些作者认为，AE 产生的聚类不如 CE 产生的聚类可靠，并且当对结核分枝杆菌复合群（MTC）分离株进行分型时，CE 比 AE 更敏感。由于这种说法可能对该领域的研究人员产生重大影响，因此对先前使用 CE VNTR 分析进行基因分型的 19 株比利时牛分枝杆菌菌株进行了比较。在 14 个 VNTR 基因座上比较了 CE VNTR 分析的 VNTR 图谱与 AE VNTR 分析的 VNTR 图谱。我们的结果表明，当将 AE VNTR 分析获得的拷贝数与 CE VNTR 分析获得的拷贝数进行比较时，所有测试基因座的拷贝数均无差异。与 CE VNTR 分析相比，AE VNTR 分析在分枝杆菌基因分型中的应用并未改变 MLVA 技术的敏感性。因此，AE VNTR 可以被视为在无法承担 CE VNTR 分析所需昂贵设备的中等设备实验室中的可行替代方法，并且可以在使用 CE VNTR 方法的实验室之间共享 AE VNTR 分析获得的数据。
