A novel rapid detection approach based on CRISPR-Cas13a for Dermatophagoides pteronyssinus and Dermatophagoides...

Dermatophagoides pteronyssinus (Trouessart) and Dermatophagoides farinae (Hughes) (Acariformes: Pyroglyphidae) are the prevalent kinds of house dust mites (HDMs). HDM is a common indoor pest, which mainly breeds in indoor dust and is an important allergen source causing a variety of allergic diseases. Effective detection of these HDMs is crucial in preventing the allergic diseases they cause. The objective of this study was to develop an innovative method for the rapid visualization of HDMs (D. pteronyssinus and D. farinae) using recombinase polymerase amplification (RPA) and lateral flow dipstick (LFD) in combination with CRISPR-Cas13a (RPA-Cas13a-LFD). To achieve heightened sensitivity in the detection of HDMs, Cas13a was incorporated into the RPA process and coupled with T7 transcripts. Based on this approach, a total of 2.23-102 copies/μl of HDM were detected within 35 min (detection limit of 2.23 copies/μl for D. farinae and 39.7 copies/μl for D. pteronyssinus). No cross-reactivity occurred with Aleuroglyphus ovatus (Troupeau) (Acariformes: Acaridae), Tyrophagus putrescentiae (Schrank) (Acariformes: Acaridae), Blomia tropicalis (van Bronswijk, de Cock & Oshima) (Acariformes: Acaridae), Suidasia nesbitti (Hughes) (Acariformes: Acaridae), and Carpoglyphus lactis(Linnaeus) (Acariformes: Acaridae). The RPA-Cas13a-LFD methodology demonstrated high specificity and sensitivity in detecting HDM. Given its advantages, such as ease of operation, rapid detection, and time efficiency, it is well-suited for rapid field-based detection of HDMs, providing a new technical tool for detecting D. farinae and D. pteronyssinus.