CAJ | 학술논문

目的利用TaqMan Real-time PCR技术,建立Kadipiro病毒(KDV)实时荧光定量PCR检测方法.方法根据GenBank发表的KDV基因序列资料分析结果,在其第12片段基因区段设计KDV特异引物与探针,利用14株不同科属病毒核酸验证方法特异性,重复实验检验方法稳定性,利用体外转录的定量RNA标准品建立基因拷贝数定量分析模型.结果引物与探针具有良好的特异性,同一样品重复检测Ct值的变异系数均＜1.8％,定量分析模型的灵敏度为100拷贝/反应.结论建立了一种特异、灵敏、高效的KDV一步法TaqMan Real-time PCR检测方法,为今后该病毒的监测与研究工作提供技术手段.
목적 이용TaqMan Real-time PCR기술,건립Kadipiro병독(KDV)실시형광정량PCR검측방법.방법 근거GenBank발표적KDV기인서렬자료분석결과,재기제12편단기인구단설계KDV특이인물여탐침,이용14주불동과속병독핵산험증방법특이성,중복실험검험방법은정성,이용체외전록적정량RNA표준품건립기인고패수정량분석모형.결과 인물여탐침구유량호적특이성,동일양품중복검측Ct치적변이계수균＜1.8％,정량분석모형적령민도위100고패/반응.결론 건립료일충특이、령민、고효적KDV일보법TaqMan Real-time PCR검측방법,위금후해병독적감측여연구공작제공기술수단.
Objective To establish a rapid and more sensitive detection method for Kadipiro virus (KDV) based on TaqMan RT-PCR.Methods Based on the KDV S12 gene sequences published in GenBank,KDV specific primers and probe were designed.The specificity and stability of the system were tested.Quantitative standard curve of KDV TaqMan RT-PCR were established.Results The specificity and stability test showed that the system is specific and the coefficient variables were all less than 1.8％.Quantitative standard curve based on the genomic copy was drawn,and the lowest detectable limit (LOD) of system is 100 copies/reaction.Conclusion TaqMan RT-PCR for KDV detection had been established,which was more sensitive and more efficient than the general PCR.