Abstract:
Dermatophagoides farina (D. farinae) and Dermatophagoides pteronyssinus (D. pteronyssinus) are the prevalent kinds of house dust mites (HDMs). HDMs are common inhalant allergens that cause a range of allergic diseases, such as rhinitis, atopic dermatitis, and asthma. The epidemiology of these diseases is associated with exposure to mites. Therefore, in the present study, a method named multiplex loop-mediated isothermal amplification (LAMP) was developed to detect environmental dust mites. The multiplex LAMP assay allows amplification within a single tube and has an ITS plasmid detection limit as low as 40 fg/µL for both single dust mites and mixed dust mites (D. pteronyssinus and D. farinae), which is up to ten times more sensitive than classical PCR techniques. Furthermore, the multiplex LAMP method was applied to samples of single dust mites and clinical dust to confirm its validity. The multiplex LAMP assay exhibited higher sensitivity, simpler instrumentation, and visualization of test results, indicating that this method could be used as an alternative to traditional techniques for the detection of HDMs.
摘要:
粉尘螨(D.farinae)和翼状螨(D.翼楼)是常见的室内尘螨(HDMs)。HDMs是常见的吸入性过敏原,会导致一系列过敏性疾病,如鼻炎,特应性皮炎,和哮喘。这些疾病的流行病学与暴露于螨虫有关。因此,在本研究中,开发了一种称为多重环介导等温扩增(LAMP)的方法来检测环境尘螨。多重LAMP测定法允许在单个管中进行扩增,并且对于单个尘螨和混合尘螨,ITS质粒检测限均低至40fg/µL(D.翼楼和D.farinae),比经典的PCR技术灵敏十倍。此外,将多重LAMP方法应用于单个尘螨和临床粉尘样品,以确认其有效性。多重LAMP分析显示出更高的灵敏度,更简单的仪器,以及测试结果的可视化,表明该方法可以用作传统的HDM检测技术的替代方法。