基因芯片技术检测生殖器溃疡性性病病原体

目的建立检测引起生殖器溃疡性性病病原体的基因芯片技术。方法设计并合成各病原体特异性探针，用点样仪将合成的特异探针点于玻片介质上，并经过点样后处理制备成基因芯片；荧光标记各病原体标准株及临床标本的特异靶基因，进行基因芯片杂交、扫描并分析结果。结果各病原体均可见特异的基因芯片荧光杂交信号：单一病原体出现一种特异荧光信号；混合病原体的不同组合出现相应组合的特异荧光信号。对40例生殖器溃疡临床标本进行芯片杂交检测，并与暗视野显微镜 聚合酶链反应(PCR)及HSV培养 PCR结果比较，具有较好的一致性(K值分别为0．882和0．947)。另外发现混合感染2例。结论本研究初步建立的基因芯片技术具有准确、特异及同时检测多重感染的特点。

Detection of Pathogens Causing Genital Ulcer Diseases Using Gene Chip Techniques

Objective To develop a gene chip for the detection of pathogens causing genital ulcer diseases (GUDs). Methods Specific probes of 4 different pathogens were designed and synthesized. Gene chip was prepared by blotting the probes onto specially treated glass slides with the use of a robotics. Target genes of standard strains for the 4 different pathogens and the clinical specimens were amplified by PCR with Cy5 fluorescence labeled primers. The labeled amplicons were hybridized with gene chips, and then scanned and analyzed using computer software. Results The fluorescence signal for specific pathogen could be found in the gene chip, illustrating that one specific fluorescence signal denoted a single pathogen, and the combination of different signals denoted the corresponding co-existence of pathogens. Examination of 40 clinical specimens obtained from 40 patients with genital ulcers with gene chip was in good concordance with dark-field microscopy plus PCR or HSV culture plus PCR, showing Kappa values of 0.882 and 0.947, respectively. In addition, mixed infections were detected in 2 specimens. Conclusion Gene chip is a sensitive method with a reliable result and it can detect multiple infections simultaneously.