title: Rapid detection of Prunus necrotic ringspot virus using magnetic nanoparticle-assisted reverse transcription loop-mediated isothermal amplification.
abstract: Prunus necrotic ringspot virus (PNRSV) has seriously reduced the yield of Prunus species worldwide. In this study, a highly efficient and specific two-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed to detect PNRSV. Total RNA was extracted from sweet cherry leaf samples using a commercial kit based on a magnetic nanoparticle technique. Transcripts were used as the templates for the assay. The results of this assay can be detected using agarose gel electrophoresis or by assessing in-tube fluorescence after adding SYBR Green I. The assay is highly specific for PNRSV, and it is more sensitive than reverse-transcription polymerase chain reaction (RT-PCR). Restriction enzyme digestion verified further the reliability of this RT-LAMP assay. To our knowledge, this is the first report of the application of RT-LAMP to PNRSV detection in Prunus species.
authors.affiliation: Key Laboratory for Fruit Biotechnology Breeding of Shandong Province, Shandong Institute of Pomology, Shandong Academy of Agricultural Sciences, Taian 271000, China.
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Zong
authors.affiliation: Key Laboratory for Fruit Biotechnology Breeding of Shandong Province, Shandong Institute of Pomology, Shandong Academy of Agricultural Sciences, Taian 271000, China.
authors.forename: Wenwen
authors.initials: W
authors.lastname: Wang
authors.affiliation: Key Laboratory for Fruit Biotechnology Breeding of Shandong Province, Shandong Institute of Pomology, Shandong Academy of Agricultural Sciences, Taian 271000, China.
authors.forename: Hairong
authors.initials: H
authors.lastname: Wei
authors.affiliation: Key Laboratory for Fruit Biotechnology Breeding of Shandong Province, Shandong Institute of Pomology, Shandong Academy of Agricultural Sciences, Taian 271000, China.
authors.forename: Jiawei
authors.initials: J
authors.lastname: Wang
authors.affiliation: Key Laboratory for Fruit Biotechnology Breeding of Shandong Province, Shandong Institute of Pomology, Shandong Academy of Agricultural Sciences, Taian 271000, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Chen
authors.affiliation: Key Laboratory for Fruit Biotechnology Breeding of Shandong Province, Shandong Institute of Pomology, Shandong Academy of Agricultural Sciences, Taian 271000, China.
authors.forename: Li
authors.initials: L
authors.lastname: Xu
authors.affiliation: Key Laboratory for Fruit Biotechnology Breeding of Shandong Province, Shandong Institute of Pomology, Shandong Academy of Agricultural Sciences, Taian 271000, China.
authors.forename: Dongzi
authors.initials: D
authors.lastname: Zhu
authors.affiliation: Key Laboratory for Fruit Biotechnology Breeding of Shandong Province, Shandong Institute of Pomology, Shandong Academy of Agricultural Sciences, Taian 271000, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Tan
authors.affiliation: Key Laboratory for Fruit Biotechnology Breeding of Shandong Province, Shandong Institute of Pomology, Shandong Academy of Agricultural Sciences, Taian 271000, China. Electronic address: qzliu001@126.com.
authors.forename: Qingzhong
authors.initials: Q
authors.lastname: Liu
mesh_terms: D052160:Benzothiazoles; D003959:Diamines; D004587:Electrophoresis, Agar Gel; D005453:Fluorescence; D019179:Ilarvirus / Q000235:genetics / Q000302:isolation & purification*; D008280:Magnetics; D008969:Molecular Sequence Data; D053758:Nanoparticles; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009930:Organic Chemicals / Q000378:metabolism; D010935:Plant Diseases / Q000821:virology; D027861:Prunus / Q000821:virology; D011804:Quinolines; D012367:RNA, Viral / Q000235:genetics; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013194:Staining and Labeling
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; D012367:RNA, Viral; C098022:SYBR Green I
keywords: Detection; PNRSV; RT-LAMP