荧光探针二次杂交技术及其在医用基因芯片中的应用

Fluorescence probe two-hybridization technique and its application in medical genechips

目的 建立一种适用于医用基因芯片的核酸检测技术。方法 运用自行设计的荧光探针 ,通过与模板 DNA和固定探针的次序杂交 ,检出特异基因片段。结果 应用该方法在基因芯片上对性病病原体菌株进行检测 ,各阳性组荧光密度值较阴性组显著下降 ;对临床标本检测的灵敏度和特异性均达到 90 %以上 ,杂交过程不需添加任何试剂 ,杂交检测时间缩短至 40分钟以内。结论 荧光探针二次杂交技术可用于医用基因芯片 ,并具有快速、简便、可靠等特点 ,有利于实现检测过程的全自动、封闭性和一体化。

Objective To establish a new nucleic acid hybridization detection technique which may be used in medical genechips. Methods The specific DNA fragment was detected by sequential two hybridization of fluorescence probe with template DNA and fixed DNA probe. Results Fluorescence probe two-hybridization (FPTH ) was applied to genechips for the detection of sex-transmitted pathogens from culture strains, and the results showed that the values of fluorescence density of the positive groups decreased remarkably when compared with those of the negative group. Both the sensitivity and specificity for detecting clinical samples are higher than 90%. There is no need of any additional reagent in hybridization procedure, and the hybridization detection can be accomplished in 40 minutes. Conclusion The FPTH technique is rapid, simple and reliable, it can also make the clinical detection process completely automatic and integrative.