摘要
目的建立基孔肯雅病毒的荧光定量PCR检测方法。方法针对基孔肯雅病毒的基因序列,设计、合成引物、探针及反应体系,摸索出最佳反应条件,建立TaqMan荧光探针法和SYBR荧光染料法。利用建立的方法对30例基孔肯雅热病标本和10份基孔肯雅病毒保存液进行扩增、检测和结果分析,并与常规RT-PCR方法的检测结果进行比对。结果实时荧光定量PCR法比常规RT-PCR法快速、灵敏。40份标本TaqMan荧光探针法、SYBR荧光染料法和常规RT-PCR方法的阳性率分别为:17.5%、17.5%、15%。统计分析表明,3种方法差异不具有统计学意义。结论实时荧光定量PCR方法快速且具有较好的特异性、敏感性、重复性,适用于基孔肯雅病毒的快速检验。
Objective To establish real time PCR(RT-PCR) detection method for Chikungunya virus.Methods Primer and FAM probe were chosen for virus gene.The samples were analyzed by TaqMan-FAM PCR and SYBR Green-based RT-PCR techniques on a fluorescence real-time PCR instrument,and the results were compared with those by conventional RT-PCR.Results TaqMan PCR and SYBR RT-PCR positive rates of Chikungunya virus were 17.5% and 17.5%;the conventional RT-PCR were 15%.These results revealed much rapid and increase sensitivity of the real-time PCR method.There were no significant differences between these methods.Conclusion The fluorescence quantitative PCR provides a more specific,more rapid and sensitive method for quantitative detection of Chikungunya virus.It is helpful for the rapid detection of Chikungunya fever.
出处
《预防医学情报杂志》
CAS
2012年第9期747-750,共4页
Journal of Preventive Medicine Information
