【摘要】 目的采用聚合酶链反应(PCR)结合反向杂交技术建立检测常见病毒性性病病原体的低密度基因芯片。方法将H IV-1,HSV-2,HPV6,HPV11的特异性寡核苷酸探针加尾并以一定阵列固定在尼龙膜上,经过处理后制成低密度芯片。在扩增过程中用B iotin-16-dUTP标记扩增产物,产物变性后分别与点有各种探针的尼龙膜反向杂交,杂交结果采用链霉亲和素-碱性磷酸酶(SP-AP)显色系统检测,并对检测体系的敏感性和特异性进行了测试。结果4种探针只与其对应的DNA杂交,而不和其他病毒杂交。敏感性试验可检出20 fg的病毒DNA。结论低密度基因芯片具有特异、敏感、快速等优点,采用低密度基因芯片一次杂交就可同时筛选鉴定多种性传播疾病(STD s)病毒的感染,值得临床推广应用。更多还原

【Abstract】 Objective To study and develop low density gene chips by using polymerase chain reaction(PCR) combining reverse hybridization technique for detection of venereal diseases caused by viruses.Methods Specific nucleotide probes of HIV-1,HSV-2 and HPV6,11 were tailed and spotted onto nylon membranes according to certain arrays,low density gene chips were thus made after being treated.DNA isolated from viruses was amplified by PCR.During these courses,amplifying products were marked by Biotin-16-dUTP.All products were denatured,then hybridized to different probes on nylon memberane.The results of hybridization were detected by SP-AP system,The sensitivity and specifity of the detection system was detected as well.Results The four probes could only hybridize with corresponding DNA,and not with other viruses.The test of sensitivity can detect 20 fg viruses’DNA.Conclusion This method of low density gene chips is specific and sensitive.It could determine multifold sexually transmitted disease(STDs) pathogens through one hybridization.Therefore it is well worth of being promoted in clinics.更多还原