Comparison of molecular protocols to detect Tomato brown rugose fruit virus in solanaceae hosts

The Tomato brown rugose fruit virus (ToBRFV) has emerged as a significant threat to Solanaceae crops, including tomato and pepper. Its presence in Mexico since 2018 has raised concerns about its impact on agricultural production. Early and accurate detection of this pathogen is essential to prevent its spread and mitigate its effects. In Mexico, several molecular techniques are employed for its diagnosis, including endpoint RT-PCR, RT-qPCR, and multiplex RT-qPCR. This research aimed to assess the efficiency of different RNA extraction methods in combination with specific PCR primers for detecting ToBRFV. Among the methods tested, the CTAB-Trizol RNA extraction protocol combined with nested PCR using primers reported by Dovas et al. (2004) was identified as the most sensitive molecular method for detecting the virus. This finding highlights the importance of selecting the appropriate combination of extraction and amplification protocols to achieve optimal sensitivity and accuracy in ToBRFV detection.