Rapid detection of clinical pathogenic bacteria and antimicrobial-resistant genes using DNA chip / 中华传染病杂志

ABSTRACT

Objective To detect the clinical pathogenic bacteria and antimicrobial-resistant genes quickly and sensitively using DNA chip.Methods Based on the analysis of 23S rRNA gene se- quences and other genes sequences associated with antimicrobial resistance(SHV＜CTX_M),oligo nucleotide microarray was designed according to different bacteria and antimicrobial-resistant genes. The DNA fragments were amplified by labeling Cy5 fluorescence and detect clinical pathogenic bacte- rias and antimicrobial-resistant genes by hybridization.Results The result of detection(10~3-10~6 bac- teria/ml)was consistent with that of some documents in domestic and overseas under ideal circum- stances of detecting bacteria genomic DNA by the Reagent Box.And it was specific and reproducible when the detection system were evaluated with some clinical isolates and drug-resistant standard strain.DNA chip could identify 16 species and 7 generics including common diverse clinical pathogenic bacteria,and could detect the drug-resistant of extended spectrum?lactamase gene simultaneously. Conclusions The methods that we have established DNA chip is a sensitive,specific and reproducable tool for supplying routine methods.