16S rDNA基因芯片检测临床常见感染性细菌

目的 提高细菌和临床微生物检测的速度和准确性，建立含有20种细菌探针在内的感染性细菌检测用基因芯片模型。方法 使用16S rDNA克隆探针和合成的寡核苷酸探针两种，利用点样仪制成基因芯片。细菌DNA经过16S rDNA通用引物扩增后与芯片上的探针杂交，然后用荧光扫描仪检测信号。结果 基因芯片能够用于细菌检测，克服探针具有广泛和灵敏的特点，但是交叉反应明显；寡核苷酸探针具有较高的特异性，但是灵敏度稍差。结论 cDNA探针和寡核苷酸探针结合或设计几个不同的探针来指向同一株细菌很可能是将来基因芯片的检测方向。

Development of 16S rDNA gene chip for detection of clinical infections bacteria

Objectivie To develop a gene chip model containing 20 probes for detection of infections bacteria.Methods The probe of 16S rDNA and 2 synthesized oligonucleutide probes were employed to make gene chips by spoter.The DNA of bacteria was amplified by 16S rDNA universal primers and the PCR product was then applied to the gene chips for hybridization.A fluorescent scanner was used to observe and record the hybridization signals.Results The strains of bacteria were correctly identified in about 8 h.The cDNA microarray was very sensitive but less specific.Conclusion The accuracy and discrimination of the gene chip can be extended by adding further microbial probes without significantly increasing complexity or cost.