16S rDNA序列在大批量细菌分离物分子鉴定中的应用研究

16S rDNA sequence as measure for identification of bacterial isolates in bulk

利用16S rDNA序列同源性分析法对保藏于中国高校工业微生物资源与信息中心(CICIMCU)的3 726株细菌分离物进行了分子鉴定。实验结果显示:其中3 073株细菌分离物可被成功鉴定至种一级水平,分属于210个种,45个属,占整个细菌分离物的82.5%;其余653株细菌分离物可鉴定到属一级,占整个细菌分离物的17.5%。研究中也发现,16S rDNA序列在芽孢杆菌属(Bacillus)和土芽孢杆菌属(Geobacillus)等少数菌属中的部分种间鉴定的敏感性不高。上述结果表明16S rDNA序列在大多数细菌种属鉴定中具有较高的特异性和敏感性,可以作为第一轮分子标识用于大批量细菌分离物的鉴定。

The feasibility of 16S rDNA sequence-based analysis for systemic classification of bacterial isolates was exten- sively evaluated. The 16S rDNA sequences of 3 726 bacterial strains collected in CICIM-CU were amplified by PCR and sequenced. The sequencing results were compared to reference data available at the GenBank database by using BLAST. Most of bacterial isolates （ 82.5% ） were assigned to species level successfully. 653 strains （ 17.5% ） were only could be assigned to genus level. Some closely-related species belonged to genus Bacillus and Geobacillus had highly similar 16S rDNA sequences, making 16S rDNA sequence analysis-based identification problematic. This study showed that 16S rDNA sequences were sufficiently sensitive for identifying most of bacterial isolates. It could be powerfully and ordinarily used as first-round molecular marker to systemic identification of bacterial isolates in bulk.