Development of a Reverse Transcription Loop-mediated Isothermal Amplification Assay for Visual Detection of Cucumber Mosaic Virus and Zucchini Yellow Mosaic Virus in Melon Seeds

doi:10.11924/j.issn.1000-6850.casb20190800499

Abstract

Abstract:

The paper aims to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid and visual detection of cucumber mosaic virus (CMV) and zucchini yellow mosaic virus (ZYMV) in melon seeds. Two sets of LAMP primers were designed according to the coat protein (CP) gene of CMV and ZYMV. Based on the optimization of reaction conditions, RT-LAMP detection method was developed and verified for specificity and sensitivity. It is suggested that the 2 kinds of LAMP method have specific amplification action respectively for CMV or ZYMV. The sensitivity for the detection of CMV and ZYMV is 10 and 100 times higher than that of the conventional PCR method. The detection result could be judged by visual color change without requiring complex equipment. The method is suitable for detection in field condition and could provide a novel rapid detection approach for CMV and ZYMV in melon seeds.

Key words: melon seeds, cucumber mosaic virus (CMV), zucchini yellow mosaic virus (ZYMV), reverse transcription loop-mediated isothermal amplification (RT-LAMP), visual detection

CLC Number:

S436.629

Yuan Yingzhe, Luo Ming, Han Jian, Pan Yanan. Development of a Reverse Transcription Loop-mediated Isothermal Amplification Assay for Visual Detection of Cucumber Mosaic Virus and Zucchini Yellow Mosaic Virus in Melon Seeds[J]. Chinese Agricultural Science Bulletin, 2020, 36(20): 107-113.

Figures/Tables 6

References 26

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名称	序列(5′→3′)
CMV-F3	TCTACCGTGTGGGTGACA
CMV-B3	GAGCACGGCGTACTTTCG
CMV-FIP	CCGTCCGCGAACATAGCAGAGTTTTGTCCGTAAAGTTCCTGCCTC
CMV-BIP	GCTGCATCTGGAGTCCAAGCCTTTTTGTCGCCAATATCAGCGC
ZYMV-F3	CTCCGGCTCAGGTGAGAA
ZYMV-B3	ATGAGACGCTCGTGTGTTG
ZYMV-FIP	GACGCGGCACAATTTTCCCATTTTTGCAGCTGTCACGAAGGAC
ZYMV-BIP	ATGTCACTGCCACGCGTGAATTTTCGGCTTATACTCCAGCAAGT

名称	序列(5′→3′)
CMV-F3	TCTACCGTGTGGGTGACA
CMV-B3	GAGCACGGCGTACTTTCG
CMV-FIP	CCGTCCGCGAACATAGCAGAGTTTTGTCCGTAAAGTTCCTGCCTC
CMV-BIP	GCTGCATCTGGAGTCCAAGCCTTTTTGTCGCCAATATCAGCGC
ZYMV-F3	CTCCGGCTCAGGTGAGAA
ZYMV-B3	ATGAGACGCTCGTGTGTTG
ZYMV-FIP	GACGCGGCACAATTTTCCCATTTTTGCAGCTGTCACGAAGGAC
ZYMV-BIP	ATGTCACTGCCACGCGTGAATTTTCGGCTTATACTCCAGCAAGT