摘要
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。
The microarray was established to detect Chikungunya virus and Sindbis virus with strong specific and high sensitivity.According to the conservative gene sequences of Chikungunya virus and Sindbis virus in GenBank,the E gene sequences of these two viruses and their primers were synthesized,and the oligonucleotide probes were designed based on the two virus,specific detection microarray was prepared.And its sensitivity,specificity and reproducibility were examined.The results showed that the sensitivity of this microarray was 100 times higher than ordinary PCR method.This microarray was specific for Chikungunya virus and Sindbis virus,which did not cross-react with other viruses such as Japanese encephalitis virus.This study established a microarray for detection of Chikungunya virus and Sindbis virus with high sensitivity and specificity,and it is suitable to epidemiological studies and species-specific detection.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2012年第10期1493-1497,共5页
Chinese Journal of Veterinary Science
基金
国家科技支撑计划资助项目(2010BAD04B02)
公益性行业(农业)资助项目(201203082)
