ABSTRACT

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.

摘要

Dermatophagoides pteronyssinus和Dermatophagoides farinae是全球家庭环境中最常见的屋尘螨 (HDM) 物种，并导致 HDM 过敏。由于 HDM 相关临床病症的流行与接触螨虫本身有关，因此在人类家庭中检测 HDM 变得越来越重要。我们的目标是开发一种快速且易于使用的多重 PCR 来检测和区分室内灰尘中的两种相关 HDM 物种。设计了新的引物，并进行了敏感性分析。还对 PCR 产物进行测序以确认该方法的特异性。两种物种的多重 PCR 检测限低至 30 pg µL ^-1. 将多重 PCR 应用于粉尘样品也导致了两种物种的高灵敏度鉴定。该协议需要少量的模板、试剂和较短的反应时间，因此为传统的 HDM 识别技术提供了替代方案。