One-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) for detection of tomato torrado virus

• DOI: 10.1007/s00705-016-2774-2
• Publication Date: 2016-02-18T00:14:00Z
• Authors: Marta Budziszewska; Przemysław Wieczorek; Aleksandra Obrępa...
• Keywords: 0301 basic medicine; Picornaviridae Infections; Base Sequence; Brief Report; Molecular Sequence Data; Picornaviridae; 3. Good health; 03 medical and health sciences; Solanum lycopersicum; Virology; RNA, Viral; Nucleic Acid Amplification Techniques; Sequence Alignment

'Torrado' disease caused by tomato torrado virus (ToTV) is responsible for considerable losses in tomato production. Therefore, a one-step reverse transcription loop-mediated isothermal amplification protocol for early and fast detection of ToTV isolates has been developed. The RNA extracted from ToTV-infected plants was tested using this protocol with a set of six primers specific for the Vp35 coat protein gene sequence. The amplified products were analyzed using amplification curves, electrophoresis, and direct staining of DNA. The sensitivity of the protocol was tenfold higher than that of conventional RT-PCR. This new protocol is inexpensive, rapid, simple, and very sensitive.