Multiplex reverse transcription loop-mediated isothermal amplification for the simultaneous detection of CVB and CSVd in chrysanthemum

doi:10.1016/j.jviromet.2014.09.008

CORC > 中国农业科学院 > 植物保护研究所

	Multiplex reverse transcription loop-mediated isothermal amplification for the simultaneous detection of CVB and CSVd in chrysanthemum
	Liu, Xing-liang 1,2; Zhao, Xi-ting 3; Muhammad, Imtiaz 1; Ge, Bei-bei 4; Hong, Bo 1
刊名	JOURNAL OF VIROLOGICAL METHODS
	2014
卷号	210 页码:26-31
关键词	Chrysanthemum Virus B Chrysanthemum stunt viroid Multiplex Reverse transcription loop-mediated isothermal amplification Restriction enzyme
ISSN号	0166-0934
DOI	10.1016/j.jviromet.2014.09.008
通讯作者	Liu, Xing-liang
英文摘要	A multiplex reverse transcription loop-mediated isothermal amplification (mRT-LAMP) assay was developed for the simultaneous detection of Chrysanthemum Virus B (CVB) and Chrysanthemum stunt viroid (CSVd), which are the major viral pathogens of chrysanthemum worldwide. Two sets of mRT-LAMP primers were designed for the coat protein gene of CVB and the complete nucleotide sequence of CSVd, and a restriction enzyme cleavage site was inserted into two pairs of species-specific primers. The mRT-LAMP assay was designed by combining these two sets fora total of eight primers. The mRT-LAMP method distinguished between CVB and CSVd due to the subsequent restriction enzyme analysis. The sensitivity of the mRT-LAMP method was 10(3) times higher than classical PCR regarding the detection limits for CVB and CSVd. No positive results were observed when RNA from other chrysanthemum pathogens were used as mRT-LAMP templates. The method was verified by testing chrysanthemum samples collected from Beijing and Henan Province and showed high reliability and sensitivity. The developed mRT-LAMP assay also offers an efficient, convenient, and rapid tool for screening chrysanthemum virus and viroid, especially CVB and CSVd, and can be diagnosed in a single reaction. These results suggest that the new mRT-LAMP method may be used routinely for virus and viroid surveys. (C) 2014 Elsevier B.V. All rights reserved.
学科主题	Biochemical Research Methods ; Biotechnology & Applied Microbiology ; Virology
语种	英语
出版者	ELSEVIER SCIENCE BV
WOS记录号	WOS:000345482700005
内容类型	期刊论文
源URL	[http://111.203.20.206/handle/2HMLN22E/11824]
专题	植物保护研究所
作者单位	1.China Agr Univ, Coll Agron & Biotechnol, Dept Ornamental Hort, Beijing 100193, Peoples R China 2.Beijing Entry Exit Inspect & Quarantine Bur, Beijing 100026, Peoples R China 3.Henan Normal Univ, Coll Life Sci, Dept Biol Sci, Xinxiang 453007, Peoples R China 4.Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Plant Dis & Insect Pests, Beijing 100193, Peoples R China
推荐引用方式 GB/T 7714	Liu, Xing-liang,Zhao, Xi-ting,Muhammad, Imtiaz,et al. Multiplex reverse transcription loop-mediated isothermal amplification for the simultaneous detection of CVB and CSVd in chrysanthemum[J]. JOURNAL OF VIROLOGICAL METHODS,2014,210:26-31.
APA	Liu, Xing-liang,Zhao, Xi-ting,Muhammad, Imtiaz,Ge, Bei-bei,&Hong, Bo.(2014).Multiplex reverse transcription loop-mediated isothermal amplification for the simultaneous detection of CVB and CSVd in chrysanthemum.JOURNAL OF VIROLOGICAL METHODS,210,26-31.
MLA	Liu, Xing-liang,et al."Multiplex reverse transcription loop-mediated isothermal amplification for the simultaneous detection of CVB and CSVd in chrysanthemum".JOURNAL OF VIROLOGICAL METHODS 210(2014):26-31.