Visual DNA diagnosis of Tomato yellow leaf curl virus with integrated recombinase polymerase amplification and a gold-nanoparticle probe.

ABSTRACT

A visual DNA diagnosis with a rapid and simple procedure has been developed on integrating recombinase polymerase amplification (RPA) and a gold nanoparticle (AuNP) probe. The entire process is implemented in only one tube with no precision instrument and requires in total 20 min to amplify a DNA fragment with RPA and to discriminate a DNA fragment with an AuNP probe. The result in various colors is directly observable with the naked eye. Through discovering a small DNA fragment of Tomato yellow leaf curl virus (TYLCV), this system can detect one copy per microlitre of virus in a pure isolate of extracted DNA and can readily identify an infected plant with a healthy appearance. This system hence provides a highly sensitive and stable DNA diagnosis. This visual method has a potential for disease diagnosis and prognostication in the field based on advantages of simplicity, high speed, portability and sensitivity.