空调隔尘网尘螨变应原基因检测

Detection of genes encoding allergens from dust mites in air filters of air conditioning systems

目的检测空调污染隔尘网尘螨变应原的基因种类。方法采集居民空调隔尘网灰尘样品20份,分别提取总基因组DNA,并以此为模板分别用粉尘螨和屋尘螨1、2、3类变应原基因(ProDerf1、ProDerp1、Derf2、Derp2、Derf3、Derp3)特异性引物PCR扩增尘螨变应原基因,PCR产物纯化后连接pUCm-T载体,并转化E.coliDH-5α感受态细胞,涂LB板(Amp+),37℃过夜培养,随机挑取单克隆菌落进行PCR验证,阳性克隆测序并进行序列比对分析。结果共20份样品,17份检测出螨性变应原基因,同时含有ProDerf1、ProDerp1、Derf2和Derp2变应原基因6份,其余阳性样品均含有粉尘螨和(或)屋尘螨1、2类变应原基因中的一种或多种。结论芜湖市区居民空调隔尘网中含有可能引起过敏性疾病的粉尘螨和屋尘螨1、2类变应原。

Objective To detect the genes encoding allergens from dust mites in the dust screens of air conditioning systems. Methods Twenty dust samples were collected from air conditioning filters in residences. After extraction of genomic DNA, genes encoding group 1, 2, and 3 allergens （ProDer f 1, ProDer p 1, Der f 2, Der p 2, Der f 3, Der p 3） of Dermatophagoides farinae and Dermatophagoides pteronyssinus were amplified with PCR using specific primers. The purified PCR products were inserted into a vector, pUCm-T, and transformed into Escherichia coli DH-5α competent cells. Transformants were plated on LB Amp＋ and incubated overnight at 37°C. A monoclonal colony was randomly selected for PCR verification, positive clone sequencing, and sequence alignment. Results Of 20 total samples, genes encoding allergens produced by dust mites were detected in 17. Of these, 6 contained ProDer f 1, ProDer p 1, Der f 2, and Der p 2; the remaining positive samples were found to contain one or more genes encoding allergens produced by D. farinae and/or D. pteronyssinus. Conclusion Air conditioner filters in human dwellings in the Wuhu area potentially contain group 1 and 2 allergens from D. farinae and D. pteronyssinus, both of which are associated with allergic diseases.