Real-time PCR-based identification methods for Megaselia scalaris (Loew) (Diptera: Phoridae) targeting mitochondrial DNA

Abstract

The scuttle fly, Megaselia scalaris (Loew, 1866) (Diptera: Phoridae), is an insect of the family Dermatidae that often poses problems as a foreign substance contaminating foods. In this study, we developed a rapid identification method for this contaminant using mitochondrial DNA based on a molecular biology technique. From the entire region of mitochondrial DNA information, a region suitable for real-time polymerase chain reaction (PCR) was chosen, and two primers and a probe specific to M. scalaris were designed. A specificity test was performed with 17 species of pests that could be contaminants in food, and the developed real-time PCR method was confirmed to show a positive result only for M. scalaris. In addition, the limit of detection of this analysis method using a 10-µL PCR solution was 1 pg of total DNA, which revealed that the method was highly sensitive.