Abstract
The tracheal mite, Acarapis woodi, is an obligate endoparasite of honeybees and a regulated pest in countries where these mites are absent. This work describes the development of a real-time PCR method for detecting tracheal mites in honeybees. The real-time PCR was evaluated for specificity, sensitivity and speed to detect A. woodi, compared to the standard manual thoracic disc method (TDM). The assay detected A. woodi down to a 1 % incidence level in bees and 1000 copies of the target DNA when using plasmid standards. Initial testing showed no cross-reaction with the other two Acarapis species from different geographical regions or with other species of mites associated with bees. However, during extensive testing of bees, a rare population of Acarapis externus mite was identified that did cross-react with the assay. Despite this cross-reaction, the assay has been shown to be a useful screening tool and results are reliable if the TDM is used as a backup to screen hives where a positive signal is obtained.
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