生物安全学报（中英文）2025，Vol.34Issue(3)：212-220,9.DOI:10.3969/j.issn.2095-1787.20240064
车前草花叶病毒外壳蛋白基因实时荧光RT-PCR检测方法的建立与应用
Development and application of RT-qPCR for detection of Plantago asiatica mosaic virus coat protein gene
摘要
Abstract
[Aim]his study aims to develop a real-time fluorescence RT-qPCR method for rapidly screening the lily Plantago asiati-ca mosaic virus(PlAMV).[Method]The specific gene coat protein(CP)sequence information of PlAMV and 16 viruses common on the same host was retrieved by NCBI Blast to detect PlAMV.The results of phylogenetic tree analysis and genetic distance analy-sis were used to determine the specific genes of PlAMV.The specific primers and probes were designed using the CP gene as a tar-get,and the specific real-time fluorescence detection method was established.The specificity of primers was verified,the sensitivity of the reaction was tested,and 21 lily samples suspected of being infected with Plantago asiatica mosaic virus were collected to test the detection effect.[Result]The results showed that the CP gene was determined as the specific gene for the detection of PlAMV,two sets of primers and probes were designed,and two sets of high-efficiency PlAMV reverse transcription fluorescence quantitative detection systems were established by using Lily samples infected with PlAMV,with the lowest virus concentration of 1 pg·μL-1,and 21 positive samples were detected by the two groups of primers and probes,with a detection rate of 100％,and the amplification efficiency of the two groups of primers and probes was similar.[Conclusion]The established RT-qPCR reaction system can specific-ally and quantitatively detect PlAMV.It can be applied to port samples for rapid and accurate screening,which provides technical support for strengthening the analysis and control of PlAMV and formulating comprehensive and systematic control measures.关键词
车前草花叶病毒/外壳蛋白/实时荧光/RT-PCR/生物信息学Key words
Plantago asiatica mosaic virus/coat protein/real-time fluorescence/RT-PCR/bioinformatics引用本文复制引用
李鑫,李扬,李辉,李可,王秀芬..车前草花叶病毒外壳蛋白基因实时荧光RT-PCR检测方法的建立与应用[J].生物安全学报（中英文）,2025,34(3):212-220,9.基金项目
国家重点研发计划项目(2021YFD1400100、2021YFD1400101) （2021YFD1400100、2021YFD1400101）
