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根据GenBank中收录的基孔肯亚病毒和辛德毕斯病毒基因的保守序列,合成2种病毒E基因序列及引物,设计针对2种病毒的寡核苷酸探针,制备基孔肯亚病毒与辛德毕斯病毒特异性检测基因芯片,并对该芯片的灵敏性、特异性和重复性进行了验证。结果显示,所建立基因芯片检测方法的灵敏度是普通PCR方法的100倍。利用所制备的基因芯片,能检测到基孔肯亚病毒和辛德毕斯病毒特异性杂交信号,阴性对照病毒(基因Ⅰ型流行性乙型脑炎病毒,基因Ⅲ型流行性乙型脑炎病毒,猪繁殖与呼吸综合征病毒及流感病毒)均无杂交信号。本试验初步建立了基孔肯亚病毒与辛德毕斯病毒特异性基因芯片检测方法,该方法灵敏度高、特异性强,适用于基孔肯亚病毒与辛德毕斯病毒的流行病学调查和种特异性鉴定。
According to the conserved sequence of Chikungunya virus and Sindbis virus contained in GenBank, two kinds of virus E gene sequences and primers were synthesized, and two kinds of virus-specific oligonucleotide probes were designed to prepare Chikungunya virus and Sindbis Virus-specific detection of gene chips, and the chip sensitivity, specificity and repeatability were verified. The results show that the sensitivity of the established gene chip detection method is 100 times that of the ordinary PCR method. Using the prepared gene chip, specific hybridization signals of Chikungunya virus and Sindbis virus, negative control virus (genotype I, Japanese encephalitis, Porcine reproductive and respiratory syndrome virus and influenza virus) have no hybridization signal. In this study, Chikungunya virus and Sindbis virus-specific gene chip detection method was initially established. The method has high sensitivity and specificity and is suitable for the epidemiological investigation and species-specificity of Chikungunya virus and Sindbis virus Identification.