A Real-Time PCR Assay for Detecting Codling Moth Cydia pomonella on Material Intercepted at U.S. Ports of Entry-A...

Codling moth Cydia pomonella is well established nearly everywhere apples are grown. Due to this almost global distribution, larvae are often intercepted at U.S. ports of entry where immature specimens cannot be identified accurately to species leading to unnecessary quarantine actions. To assist with identifying intercepted C. pomonella from port inspections, we developed a probe-based real-time PCR assay to amplify the internal transcribed spacer (ITS) region 2 of C. pomonella. The assay was tested for inclusivity using 110 C. pomonella specimens from six continents. Analytical specificity was examined by testing related species intercepted at U.S. ports of entry, as well as non-targets with the same geographic distribution and host species as C. pomonella. The assay developed here identified all C. pomonella individuals correctly and produced appropriately negative results for all non-target species. These results ensure that the assay provides a rapid and accurate tool for unambiguously identifying C. pomonella among material intercepted at U.S. ports of entry. Since C. pomonella is not actionable, the ability to identify all life stages of C. pomonella conclusively will save time, effort, and money while also directing identification efforts towards species of current quarantine concern.