Anal Chem. 2019 Apr 2;91(7):4649-4655. doi: 10.1021/acs.analchem.8b05984. Epub 2019 Mar 11.
Capillary zone electrophoresis (CZE) can produce high-resolution separations of biological samples, including microbial mixtures. The study of complex populations of microorganisms using CZE is limited because most detectors have limited sensitivity, are destructive, and provide limited information for microbial identification. To address these issues, we developed an integrated capillary zone electrophoresis apparatus to fractionate bacteria from complex mixtures. We deposited fractions onto nutrient agar in a Petri dish for microbial culturing, and we subjected the observed colonies to Sanger sequencing of a phylogenetic marker, the 16S rRNA gene, for microbial identification. We separated and cultured both a single bacteria species, the model Gram-negative organism Escherichia coli, and a complex environmental isolate of primary sewage effluent. Sequence analysis of the 16S rRNA genes from this mixture identified 15 ± 5 distinct bacterial species per run. This approach requires minimal manipulation of microbial populations and combines electrophoretic fractionation of bacterial cells with automated collection for accurate identification of species. This approach should be applicable to microorganisms in general and may enable discrimination of physiologically different cells in complex assemblages, such as in microbiome samples.
毛细管区带电泳（CZE）可以对生物样品进行高分辨率分离，包括微生物混合物。由于大多数检测器的灵敏度有限、具有破坏性，并且只能提供有限的微生物鉴定信息，因此使用 CZE 研究复杂的微生物种群受到限制。为了解决这些问题，我们开发了一种集成的毛细管区带电泳装置，用于从复杂混合物中分离细菌。我们将馏分沉积在培养皿中的营养琼脂上进行微生物培养，并对观察到的菌落进行微生物鉴定的系统发育标记物 16S rRNA 基因的 Sanger 测序。我们分离和培养了单一细菌物种，即模式革兰氏阴性菌大肠杆菌，以及原生污水的复杂环境分离物。从该混合物的 16S rRNA 基因序列分析中，每个运行可鉴定出 15±5 种不同的细菌物种。该方法对微生物种群的操作要求最小，将细菌细胞的电泳分离与自动化收集相结合，可准确鉴定物种。该方法应该适用于一般的微生物，并且可以区分复杂组合中生理上不同的细胞，例如在微生物组样本中。
