用聚合酶链反应技术对粉尘螨和屋尘螨进行鉴别和检测

Identification and detection of Dermatophagoides farinaand Dermatophagoides pteronyssinus by Polymerase Chain Reaction

目的用聚合酶链反应(PCR)技术对粉尘螨和屋尘螨进行鉴别和检测。方法用随机引物PCR对粉尘螨和屋尘螨的DNA进行扩增,选择3个扩增条带进行测序和分析。根据测序结果再次设计PCR引物用于粉尘螨和屋尘螨的鉴别和检测,并对该引物的特异性和敏感性进行了分析。结果随机引物扩增反应,粉尘螨在750bp和500bp左右均有明显条带;而屋尘螨仅500bp左右条带较清晰,无大于500bp的扩增产物出现。测定了3个新的尘螨DNA片段的序列结构。新设计的引物用于两种尘螨的检测有较好的特异性和敏感性。结论随机引物PCR技术可用于粉尘螨和屋尘螨的鉴别。本研究方法具有检测环境中粉尘螨和屋尘螨的潜力。

The purpose of the present study was to identify and detect Dermatophagoides farina（D.f） and Dermatophagoides pteronyssinus（D.p）by Polymerase Chain Reaction（PCR） technology.A randomly primed PCR（RP-PCR） technology was used to amplify the DNA of D.f and D.p,and three amplified bands were selected for DNA sequencing and analysis.According to the resuLts of DNA sequencing,another two pairs of primers were designed to identify and detect D.f and D.p,then the specificity and sensitivity of these primers were analyzed.In amplification of RP-PCR,two significant bands around 500bp and 750bp were observed in amplified products of D.f,while only a clear band about 500bp in D.p and no product larger than 500bp appeared.Three new DNA fragments of D.f and D.p were sequenced and analyzed in homology.As the new primers designed were highly specific and sensitive in detection of the two mites,RP-PCR technology could be used to identify D.f and D.p.Results indicate that the PCR method verified in the research has a potentiality to detect D.f and D.p in the environment.