Mite species identification in the production of allergenic extracts for clinical use and in environmental samples by ribosomal DNA amplification

Abstract

The identification of allergy-causing mites is conventionally based on morphological characters. However, molecular taxonomy using ribosomal DNA (rDNA) may be particularly useful in the analysis of mite cultures and purified mite fractions in the production of allergenic extracts. Full-length internal transcribed spacers (ITS1 and ITS2) were obtained from Dermatophagoides farinae, Dermatophagoides pteronyssinus, Dermatophagoides microceras and Euroglyphus maynei (Astigmata: Pyroglyphidae), Glycyphagus domesticus and Lepidoglyphus destructor (Astigmata: Glycyphagidae), Tyrophagus fanetzhangorum, Tyrophagus putrescentiae, Tyrophagus longior, Tyrophagus neiswanderi, Acarus farris and Acarus siro (Astigmata: Acaridae), and Blomia tropicalis (Astigmata: Echymopodidae), using mite-specific primers. Polymerase chain reaction (PCR) products were digested with HpaII and RsaI restriction enzymes in order to produce species-specific PCR restricted fragment length polymorphism (RFLP) profiles. A semi-nested re-amplification step was introduced before the RFLP in order to apply the method to environmental samples. Results demonstrate that rDNA sequences can be used for the unambiguous identification of mite species. The PCR-RFLP system allows the identification of species in purified mite fractions when the availability of intact adult mite bodies for morphological identification is limited. This reliable and straightforward PCR-RFLP system and the rDNA sequences obtained can be of use in the identification of allergy-causing mite species.

Allergens of <b><i>Blomia tropicalis</i></b>: An Overview of Recombinant Molecules

“…Nowadays it is well accepted that B. tropicalis also constitutes a very important house dust mite species, especially, but not exclusively, in the tropical and subtropical regions of the world [2,3,4,5]. In addition to allergen polymorphisms, some authors have claimed seasonal dynamic patterns of mite species, allergen densities within them, and different specific breeding sites [6,7,8,9,10]. All of these factors influencing the allergen prevalence could in turn have a direct effect on the allergenicity of mites within different environments and/or periods of the year.…”

The influence of household pets on the composition and quantity of allergenic mite fauna within Irish homes: a preliminary investigation

“…domesticus,L. destructor and T. putrescentiae (Table 2) are all known allergen producers (Solarz et al, 2004;Skelton et al, 2007;Colloff, 2009;Celebioglu et al, 2013;Beroiz et al, 2014). Chortoglyphus arcuatus has been studied in relation to its allergenicity in regions such as Colombia (Puerta et al, 1993), Spain (López-Conde et al, 2005;Boquete et al, 2006) and Poland (Solarz et al, 2004).…”

An Integrative Taxonomy of Molecular and Traditional Approaches for Identification of a Storage Mite, Aleuroglyphus Ovatus (Acari: Astigmata: Acaridae) in Malaysia

“…This method allows identification for immature of the target species and broken mites (Wong et al, 2011), which cannot be identified morphologically. Furthermore, this method can be applied to DNA extracted from purified fractions of bodies and faeces which can be used as source materials in the production of allergenic extracts (Beroiz et al, 2014). One possible reason for the faint bands and decline of visualization of PCR products signal below 4 µl may be due to the increasing insensitivity of ITS2 gene amplification for detecting a smaller amount of mite's DNA in a less concentrated dilution.…”

“…c o m allergic diseases, such as asthma, rhinitis and atopic dermatitis (Silton et al 1991). Species identification in mite cultures is a key factor in standardization of allergen production especially when the colonies established are from field-derived mites (Beroiz et al, 2014). Proper species identification may contribute to improved management of mite associated allergies as they produce species-specific allergens besides being an essential first step of acarological evidence for a legal investigation (de Pancorbo et al, 2004;Yang et al, 2011).…”