检测产超广谱β-内酰胺酶耐药基因寡核苷酸芯片的研制及应用

Development and application of oligonucleotide microarray in detecting extended spectrum β-lactamases drug resistance genes

目的探讨用寡核苷酸芯片检测革兰阴性杆菌耐产超广谱β-内酰胺酶抗生素耐药基因的可行性。方法制备了检测产超广谱β-内酰胺酶革兰阴性杆菌寡核苷酸芯片。应用该寡核苷酸芯片检测16例临床革兰阴性杆菌耐药株,并与测序结果比较。结果提取的DNA产物随机引物标记后杂交,杂交结果差异无统计学意义。其结果与测序结果相符。结论寡核苷酸芯片可以推广应用到临床作为耐产超广谱β-内酰胺酶抗生素快速诊断的一种有效方法。

Objective To explore the feasibility of detecting the extended spectrum β-lactamases drug resistance genes in Gram negative bacteria by means of oligonucleotide microarray. Methods Probes were designed and the gene chip was fabricated according to the sequences of the16 antimicrobial resistance gene cassette published in Genebank. And 16 Gram-negative strains were detected by using DNA microarray for identification drug resistance. Hybridization between probes and genomic DNA and the results were compared with the sequence. Results As a result,it is conistent with the one expected before. Conclusion This research implies that oligonucleotide microarray can be applied into the quick diagnosis of the extended spectrum β-lactamases drug resistance genes in Gram-negative bacteria