The Use of Real-Time PCR to Identify Blueberry Maggot (Diptera: Tephritidae, Rhagoletis mendax) from Other Rhagoletis Species and in Lowbush Blueberry Fruit (Vaccinium angustifolium)

Blueberry maggot is the key insect pest of lowbush blueberry in Atlantic Canada. Growers must ensure fruit is free of larvae as there is zero tolerance for them in exported fruit; it is difficult to confirm this. Real-time PCR is a rapid and efficient method for identification of insect species. Seven sets of PCR primers found in the Cytochrome Oxidase I and II regions were tested with DNA from the four life stages of three Rhagoletis spp. PCR products were sequenced and the intra-specific and inter-specific similarities were 100% and 91-99.5%, respectively. Four species-specific primer sets were designed for real-time PCR using SYBR Green and melting curve analysis. This technique was found to be reliable, fast and able to detect and identify Rhagoletis spp. from the fruit of lowbush blueberry.