U.S. flag

An official website of the United States government

Dot gov

Official websites use .gov
A .gov website belongs to an official government organization in the United States.


Secure .gov websites use HTTPS
A lock ( ) or https:// means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.


Main content area

Development of loop-mediated isothermal amplification assay for rapid detection of cucurbit Leaf crumple virus

Waliullah Sumyya, Ling Kai Shu, Cieniewicz Elizabeth J., Oliver Jonathan E., Ji Pingsheng, Ali Emran
International journal of molecular sciences 2020 v.21 no.5 pp. -
Cucurbit leaf crumple virus, Cucurbitaceae, DNA primers, detection limit, disease detection, foliar diseases, gel electrophoresis, loop-mediated isothermal amplification, rapid methods, signs and symptoms (plants), viral diseases of plants
A loop-mediated isothermal amplification (LAMP) assay was developed for simple, rapid and efficient detection of Cucurbit leaf crumple virus (CuLCrV), one of the most important begomovirus that infects cucurbits worldwide. A set of six specific primers targeting a 240 nt sequence region in the DNA A of CuLCrV were designed and synthesized for detection of CuLCrV from infected leaf tissues by real-time LAMP amplification using Genie® III system, which was further confirmed by gel electrophoresis and SYBR Green I DNA staining for visual observation. The optimum reaction temperature and time were determined, and no cross-reactivity was seen with other begomoviruses. The sensitivity assay revealed that LAMP was more sensitive than conventional PCR but less sensitive than qPCR. However, it was simpler and faster than the other assays evaluated. LAMP assay was also more efficient and sensitive in detecting CuLCrV from symptomatic and asymptomatic infected field samples than PCR. This simple, rapid, and sensitive method has the capacity to detect CuLCrV in samples collected in the field and is therefore suitable for early detection of the disease to reduce the risk of epidemics.