Development of a new multiplex PCR to detect prevalent species of house dust mites in house dust

Oliveira, Ana Sofia; Gaspar, Carlos; Rolo, Joana; Pereira, Cristiana Costa; Palmeira-de-Oliveira, Rita; Teixeira, João Paulo; Martinez-de-Oliveira, José; Palmeira-de-Oliveira, Ana

http://hdl.handle.net/10400.18/8598

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Autores

Oliveira, Ana Sofia

Gaspar, Carlos

Rolo, Joana

Pereira, Cristiana Costa

Palmeira-de-Oliveira, Rita

Teixeira, João Paulo

Martinez-de-Oliveira, José

Palmeira-de-Oliveira, Ana

Resumo(s)

Dermatophagoides pteronyssinus and Dermatophagoides farinae are the most common House Dust Mite (HDM) species in home environments worldwide and responsible for HDM allergy. Since the prevalence of HDM-related clinical conditions is linked to exposure to the mite itself, the detection of HDM in the human households gains importance. We aimed to develop a fast and accessible multiplex PCR to detect and distinguish two relevant HDM species in house dust. New primers were designed, and sensitivity analysis was performed. Sequencing of PCR products was also performed to confirm the method's specificity. The limit of detection of the multiplex PCR for both species was as low as 30 pg µL-1. The application of the multiplex PCR to dust samples also resulted in the identification of both species with high sensitivity. The protocol required small amount of template, reagents and a short reaction time thus presenting an alternative to classically used techniques for HDM identification.