基孔肯雅病毒和辛德毕斯病毒可视化基因芯片与荧光基因芯片的建立

根据GenBank中收录的基孔肯雅病毒和辛德毕斯病毒E蛋白基因序列,设计及筛选针对2种病毒的寡核苷酸探针及引物,制备基孔肯雅病毒与辛德毕斯病毒可视化基因芯片与荧光基因芯片,对芯片的灵敏性、特异性进行了验证,并将可视化基因芯片、荧光基因芯片进行灵敏性比较.结果显示,制备的两种基因芯片都能检测到基孔肯雅病毒和辛德毕斯病毒特异性杂交信号.可视化基因芯片、荧光基因芯片检测两种病毒质粒的灵敏度达到9.1×103copies/mL,6.8×101copies/mL和9.1×104copies/mL,6.8×103copies/mL,与普通PCR比较差异显著.荧光基因芯片灵敏度是PCR方法的10倍,可视化基因芯片是荧光基因芯片灵敏度的100倍.模拟病毒检测过程特异性检验证明,可视化基因芯片都具有良好的特异性.本试验建立了基孔肯雅病毒与辛德毕斯病毒两种特异的可视化和荧光基因芯片检测方法,两种方法灵敏度高、特异性强,适用于基孔肯雅病毒与辛德毕斯病毒的流行病学调查和种特异性鉴定.

Establishment of the Visualized Microarrays and the Fluorescence Microarrays for Chikungunya Virus and Sindbis Virus

According to the conservative E gene sequences of Chikungunya virus and Sindbis virus from GenBank, the oligonucleotide probes and primers were designed. The fluorescence and visualized specific detective gene chips were prepared and their sensitivity, specificity were examined. Comparison of the sensitivity among the ordinary PCR and the two kinds of gene chips has been made. The distinction of the sensitivity between the gene chips and the ordinary PCR is obvious, while the fluorescence gene chip is 10-fold higher than the PCR. The sensitivity of the visualized gene chip and the fluorescence microarray for Chikungunya virus and Sindbis virus are 9.1×103 copies/mL, 6.8×101 copies/mL and 9.1×104 copies/mL, 6.8×103 copies/mL and the former is 100-fold higher than the later. Through the procedure of modeling the viral special detection, it has been proved that the visualized gene chip methods have a good specificity. This study has established two kinds of microarrays for detection of Chikungunya virus and Sindbis virus with high sensitivity and specificity. It is suitable for epidemiological studies and species-specific detection.