recA基因及核糖体分型技术在洋葱伯克霍尔德菌复合体鉴别与溯源分析中的应用

目的：对药品中分离的疑似洋葱伯克霍尔德菌复合体（BCC）进行准确鉴别及溯源分析。方法：通过核糖体自动分型对不同来源分离菌株进行核糖体型分类，选取各型代表菌株（8株）进行生化实验、16SrRNA及recA基因序列测定和同源性分析，确定其在菌种水平的分类地位。结果：核糖体自动分型可针对实验中分离的菌株进行核糖体型的聚类区分；对3类核糖体型的代表菌株进行的生化鉴定、16S rRNA序列分析将其定位至洋葱伯克霍尔德菌复合体；recA基因序列的同源性分析对待测菌株进行种水平鉴别，印证核糖体分型结果并将待测菌株鉴定为洋葱伯克霍尔德菌、新洋葱伯克霍尔德菌及稳定伯克霍尔德菌。结论：recA序列同源性分析可对样品中分离的洋葱伯克霍尔德菌复合体（BCC）在种水平上进行有效鉴别，核糖体分型可对不同来源BCC进行溯源，确定污染来源。

Objective: To carry out an accurate identification and traceability analysis of suspected Burkholderia cepacia complex (BCC)isolated from drugs. Methods: By using automated ribotyping, the ribotypes of several strains isolated from different sources were distinguished. A polyphasic taxonomic approach including automated biochemical screening, 16S rRNA and recA gene sequencing were used to determine the taxonomic position of eight typical stains. Results: All isolated strains were seperated into three kinds of ribotypes, and eight of them were identified as BCC by using automated biochemical screening and 16S rRNA sequence homology analysis. Through phylogenetic analysis of recA gene, eight isolated strains were resolved at the species level:four Burkholderia cepacia (genomovarⅠ), two Burkholderia cenocepacia (genomovar Ⅲ), and one Burkholderia stabilis (genomovar Ⅳ). Conclusion: The homology analysis of specified housekeeping gene recA can effectively identify BCC isolates, and ribotyping is an effective method for traceability analysis of BCC to determine the source of contamination.

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