Rapid diagnosis of two common stored-product predatory mite species based on species-specific PCR

Cheyletus malaccensis Oudemans and Cheyletus eruditus (Schrank) are two sibling species of the Cheyletus genus. Both species are beneficial predators of stored product pests. It is difficult to distinguish them based only on morphological characteristics. In this study, we describe species-specific PCR for molecular identification of C. malaccensis and C. eruditus. Species-specific primer pairs were designed on the basis of variations in the mtDNA COI barcode sequences. A 403 bp-specific fragment of C. malaccensis and a 421 bp absolute product of C eruditus were generated by their respective primer pairs, while no such diagnostic bands were present in any of the nine pest mite species tested. The results of the sensitivity test demonstrated that the detection limit of DNA template concentration was 1 ng/mu L for both C malaccensis and C eruditus. This study exhibited high specificity, reliability, rapid and low cost for mite specie diagnosis. Our data show that species-specific PCR is a powerful tool that can be used for identification of C malaccensis and C eruditus. (C) 2016 Elsevier Ltd. All rights reserved.