LAMP and real-time PCR assays for rapid identification of Frankliniella intonsa

Abstract

Frankliniella intonsa Trybom is a pest causing significant economic losses of crops cultivated in greenhouses and fields, especially losses of ornamental and horticultural plants. This species is responsible for causing direct damage to plants as well as for transmitting viruses, including the tomato spot wilt virus (TSWV), which is considered the most harmful. Because the proper identification of a pest species is crucial for plant protection, and morphological identification methods require good entomological expertise and are time consuming, molecular diagnostic techniques are a great alternative. Among the molecular diagnostic protocols available for F. intonsa detection, no protocols based on the loop-mediated isothermal amplification (LAMP) method or real-time PCR have been described thus far. Therefore, the aim of this study was to develop species-specific, fast and sensitive protocols based on these two approaches for the identification of F. intonsa. The real-time PCR and LAMP assays developed and described here gave positive results only for F. intonsa populations with a thrips DNA concentration as low as 10−3 ng/μl, proving that they are species specific and sensitive. No reaction products were observed for the negative controls. Moreover, the described LAMP protocol allows for the identification of F. intonsa without a DNA extraction step.