Development of a sensitive real-time quantitative RT-PCR assay for the detection of pear chlorotic leaf spot-associated virus

被引：1
|
作者：
Zhu, Yiting [1 ]
Hong, Ni [1 ]
Li, Liu [1 ]
Gao, Yujie [1 ]
Wang, Liping [1 ]
Xu, Wenxing [1 ]
Wang, Guoping [1 ]
机构：
[1] Huazhong Agr Univ, Coll Plant Sci & Technol, Key Lab Plant Pathol Hubei Prov, Wuhan 430070, Hubei, Peoples R China
来源：
JOURNAL OF VIROLOGICAL METHODS | 2022年 / 309卷
关键词：
Pear; Pear chlorotic leaf spot -associated virus; Reverse transcription quantitative PCR; RT-PCR;
D O I：
10.1016/j.jviromet.2022.114608
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
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页数：7
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