基于高通量测序技术分析特应性皮炎患者皮肤微生物菌群多样性

Analysis of skin microbiota diversity in patients with atopic dermatitis based on high throughput sequencing

目的 采用16S rDNA、ITS测序技术研究特应性皮炎患者皮肤微生物菌群多样性。方法 收集本院25例特应性皮炎患者(实验组)及22例健康人(正常对照组),取皮损处(PL组)及无皮损处(PN组)皮肤菌群样本,正常对照组取相应部位菌群样本(N组)。提取样品DNA行illumina高通量测序。结果 基于16S rDNA多样性指数分析显示,PL组shannon指数低于PN、N组(均P<0.01),simpson指数低于PN、N组(均P<0.01),表明特应性皮炎患者皮损处细菌菌群多样性显著性降低。3组间β多样性差异有统计学意义(P<0.05),3组间菌群组成差异有统计学意义(P=0.001)。同时,ITS多样性指数分析显示特应性皮炎患者皮损处真菌菌群多样性升高。结论 通过16S rDNA以及ITS测序技术应用于特应性皮炎皮肤微生物组学研究,证实了特应性皮炎患者皮肤菌群多样性与正常皮肤存在一定差异,皮肤表面微生物菌群与特应性皮炎的发病密切相关。

Objective To analyze the skin microbiota diversity in patients with atopic dermatitis(AD)using 16S rDNA and ITS sequencing.Methods Twenty-five patients with AD and 22 healthy controls were collected from the Dermatology Hospital of Fuzhou.Skin swabs were collected from lesional skin(PL group)and non-lesional skin(PN group)of the patients with AD, as well as from the corresponding body sites of the healthy controls(N group).DNA was extracted from the samples and subjected to high throughput sequencing for diversity index analysis.Results 16S rDNA diversity index analysis showed that shannon index in PL group was significantly lower than PN and N group(P<0.01) with simpson index higher in PN and N group(P<0.01),indicating that the diversity of bacterial flora in skin lesions of AD patients decreased substantially.There was significant difference in the composition of microflora among the three groups(P=0.001).Meanwhile, ITS diversity analysis showed that the diversity of fungal flora in the skin lesions of AD patients increased.Conclusion 16S rDNA and ITS sequencing technology were applied to study the microbiome of atopic dermatitis.The results of this study confirmed that the diversity of AD skin bacteria were different from that of normal people.The microflora of skin surface is closely related to the pathogenesis of atopic dermatitis.