猪链球菌基因芯片检测方法的研究

Study on oligonucleotide microarray to detect of Streptococcus suis

目的建立猪链球菌基因芯片检测方法,实现对猪链球菌种、致病血清型及毒力相关基因的同步检测和鉴定。方法根据GenBank中猪链球菌的相关序列,设计并合成探针,采用点样法制备芯片;检测样本基因组DNA标记后与基因芯片杂交,激光扫描检测信号。结果检测结果显示,针对猪链球菌种、主要致病血清型及主要毒力相关基因设计的寡核苷酸探针可特异地识别对应靶基因,重复性好;而针对不同菌株设计的株特异性探针未获得预期检测效果。结论初步建立猪链球菌基因芯片检测体系,敏感性与特异性高,对于高通量鉴定猪链球菌菌种及其毒力有重要价值。

In order to develop a DNA microarray to detect Steptococcus suis species, its serotype and virulence-associated genes simultaneously, a set of probes specific for the target genes were designed. These probes were synthesized by DNA synthesizer. The microarray was prepared by spotting these probes onto the specially treated glass sliders. Genomic DNA of specimen was prepared routinely and labeled. Following hybridization of labeled products on the microarrays, detection was carried out by fluorescence scanner. It was demonstrated that the probes designed for Steptococcus suis species, serotype and virulenceassociated genes could detect the corresponding target genes with a great sensitivity and specifity,however detection with probe specific for different strains failed. It is apparent that the diagnostic genechip method is sensitive and specific to detect pathogenic Streptococcus suis.