Presentation Title

Detection of Tomato Mosaic Virus Using a Novel at Home RT-PCR Approach

Presentation Type

Presentation

Full Name of Faculty Mentor

Michelle Barthet, Biology

Major

Biology

Presentation Abstract

Tomato mosaic virus (ToMV) belongs to the genus Tobamovirus consisting of positive-strand RNA viruses. ToMV has limited tomato production globally. Current testing methods require expensive molecular equipment for high complexity molecular diagnostics and are limited to a laboratory environment. Novel ToMV primers targeted against ToMV genes 2 and 4 were created for detection of ToMV in tomato leaves and were checked by BLAST analysis in GenBank for non-specificity to other viruses. A modified RNA extraction protocol and a simplified RT-PCR protocol with SYBR green was used to amplify and detect ToMV in a manner for replication in a kitchen environment. An internal control was incorporated to confirm positive results and successful RNA extraction. Initial tests demonstrated the efficacy of the methodology. This RNA extraction and RT-PCR approach will help detect ToMV outside of the laboratory and lead to detection methods for various viruses when materials and facilities are lacking.

Location

Room 1

Start Date

21-4-2021 5:10 PM

End Date

21-4-2021 5:30 PM

Disciplines

Biology

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Apr 21st, 5:10 PM Apr 21st, 5:30 PM

Detection of Tomato Mosaic Virus Using a Novel at Home RT-PCR Approach

Room 1

Tomato mosaic virus (ToMV) belongs to the genus Tobamovirus consisting of positive-strand RNA viruses. ToMV has limited tomato production globally. Current testing methods require expensive molecular equipment for high complexity molecular diagnostics and are limited to a laboratory environment. Novel ToMV primers targeted against ToMV genes 2 and 4 were created for detection of ToMV in tomato leaves and were checked by BLAST analysis in GenBank for non-specificity to other viruses. A modified RNA extraction protocol and a simplified RT-PCR protocol with SYBR green was used to amplify and detect ToMV in a manner for replication in a kitchen environment. An internal control was incorporated to confirm positive results and successful RNA extraction. Initial tests demonstrated the efficacy of the methodology. This RNA extraction and RT-PCR approach will help detect ToMV outside of the laboratory and lead to detection methods for various viruses when materials and facilities are lacking.

https://digitalcommons.coastal.edu/ugrc/test1/test1track/75