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Detection of melon necrotic spot virus by one-step reverse transcription loop-mediated isothermal amplification assay.

ABSTRACT: Melon necrotic spot virus (MNSV) can cause significant economic losses due to decreased quality in cucurbit crops. The current study is the first to use reverse transcription loop-mediated isothermal amplification (RT-LAMP) for detection of MNSV. A set of four LAMP primers was designed based on the coat protein gene sequence of MNSV, and a RT-LAMP reaction was successfully performed for 1 h at 62°C. The results of RT-LAMP showed high specificity for MNSV and no cross-reaction with other viruses. Compared to traditional reverse transcription-PCR (RT-PCR), the RT-LAMP assay was 103-fold more sensitive in detecting MNSV. Due to its sensitivity, speed and visual assessment, RT-LAMP is appropriate for detecting MNSV in the laboratory.

SUBMITTER: Qiao N

PROVIDER: S-EPMC7058275 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Detection of melon necrotic spot virus by one-step reverse transcription loop-mediated isothermal amplification assay.

Qiao Ning N Dai Huijie H Liu Jie J Zhu Xiaoping X Li Jintang J Zhang Dezhen D Liu Yongguang Y

PloS one 20200305 3

Melon necrotic spot virus (MNSV) can cause significant economic losses due to decreased quality in cucurbit crops. The current study is the first to use reverse transcription loop-mediated isothermal amplification (RT-LAMP) for detection of MNSV. A set of four LAMP primers was designed based on the coat protein gene sequence of MNSV, and a RT-LAMP reaction was successfully performed for 1 h at 62°C. The results of RT-LAMP showed high specificity for MNSV and no cross-reaction with other viruses. ...[more]

PMID: 32134962

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