梨褪绿叶斑相关病毒的灵敏实时荧光定量 RT-PCR 检测方法的建立。
Development of a sensitive real-time quantitative RT-PCR assay for the detection of pear chlorotic leaf spot-associated virus.
机构信息
The Key Laboratory of Plant Pathology of Hubei Province, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
The Key Laboratory of Plant Pathology of Hubei Province, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
出版信息
J Virol Methods. 2022 Nov;309:114608. doi: 10.1016/j.jviromet.2022.114608. Epub 2022 Aug 25.
Pear chlorotic leaf spot associated virus (PCLSaV) belongs to the genus Emaravirus and possesses a genome composed of five negative-sense single-stranded RNA (-ssRNA) segments. This study developed a SYBR green-based reverse transcription quantitative PCR (RT-qPCR) assay for the detection of PCLSaV infecting pear trees. A set of two primers q5-F2/q5-R2 designed based on the viral RNA5 sequences showed high specificity and feasibility for PCLSaV detection. The standard curve was established. RT-qPCR assays showed that PCLSaV content was greatly higher in diseased branch and symptomatic leaf samples than that in un-diseased branch and asymptomatic leaf samples. The RT-qPCR was reliability in the detection of the virus in field and in-vitro cultured pear samples. This technique would be useful for the supervision of the viral disease and the certification of pear planting materials.
梨褪绿叶斑相关病毒（PCLSaV）属于埃玛病毒属，其基因组由五个负义单链 RNA（-ssRNA）片段组成。本研究开发了一种基于 SYBR Green 的逆转录定量 PCR（RT-qPCR）检测方法，用于检测感染梨树的 PCLSaV。根据病毒 RNA5 序列设计的一对引物 q5-F2/q5-R2 对 PCLSaV 的检测具有高度的特异性和可行性。建立了标准曲线。RT-qPCR 检测结果表明，病枝和病叶样本中的 PCLSaV 含量明显高于健康枝和无症状叶样本。该 RT-qPCR 方法可用于田间和体外培养的梨样品中病毒的检测。该技术将有助于对病毒病的监测和梨种植材料的认证。
Zotero 插件