摘要
目的基于Illumina MiSeq测序平台,分析血液透析水处理系统水样中微生物多样性和丰度。方法采集血液透析水处理设备供水处自来水、活性碳罐后软化水、出水口反渗水水样各两份,分为Ⅰ、Ⅱ两个组别,Ⅰ组水样进行传统微生物培养;Ⅱ组水样进行二代测序。结果Ⅰ组经微生物培养细菌总数均不超标,质谱鉴定为假单胞菌种;Ⅱ组经二代测序,获取可操作分类单元(OTU)总数依次为3 957,3 245,4 880,分别来自428,494和369个微生物类群。反渗水水样在门、纲、目、科、属的OTU数均明显高于自来水和软化水水样;其Chao1和ACE丰富度估计指数最高,表明群落中实际存在的物种数最多。分类组成分析反渗水水样从科、属分类上约58.92%为假单胞菌科,58.90%为假单胞菌属;26.41%为莫拉菌科,26.23%为莫拉菌属;4.73%为普氏菌属。结论二代测序技术反映了血液透析水处理系统水样中微生物的多样性和丰度,传统微生物培养方法的局限性使其严重低估了反渗水中细菌数量及菌群结构,使透析用水污染引起透析患者微炎症反应被忽视。
OBJECTIVE To investigate the microbial diversity and abundance in hemodialysis water treatment system based on Illumina MiSeq sequencing platform. METHODS The tap water of hemodialysis water treatment equipment, softened water after the activated carbon tank and reverse seepage at the outlet were collected for each two copies and divided into the group Ⅰ and the group Ⅱ. The group Ⅰ was treated with conventional microbial culture, and the group Ⅱ was treated with second generation sequencing. RESULTS The result of microbial culture showed that the total bacterial colony counts of the group Ⅰ did not exceed the standard, and the strains were identified as the Pseudomonas by mass spectrometry. The result of second generation sequencing showed that the total colony counts of Optical Transform Unit(OUT) were respectively 3957,3245 and 4880,which were respectively from 428,494 and 369 microbial groups. The OTU counts of the reverse osmosis water samples were remarkably greater than those of the tap water and softened water samples in phylum, class, order, family and genus;Chao1 and ACE of the reverse osmosis water samples were the highest, indicated that there were largest number of species in the microbial flora. The result of bacterial classification on family and genus level analysis showed that of the strains isolated from the reverse osmosis water samples, 58.92% were Pseudomona daceae,58.90% of which were Pseudomonas;26.41% were Moraxellaceae,26.23% of which were Moraxella;4.73% were Prevotella. CONCLUSION The second generation sequencing technology reflects the diversity and abundance of microorganisms in water of hemodialysis water treatment system. The limitations of traditional microbial culture methods have seriously underestimated the bacterial colony counts and the structure of flora, and the microinflammatory response induced by contamination of dialysis water has been neglected.
作者
绽丽
杨冬华
刘佳雯
张永栋
赵璐
ZHAN Li;YANG Dong-hua;LIU Jia-wen;ZHANG Yong-dong;ZHAO Lu(Qinghai University Affiliated Hospital , Xining , Qinghai 810001, China)
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2019年第13期2063-2068,共6页
Chinese Journal of Nosocomiology
基金
青海大学附属医院中青年科研基金资助项目(ASRF-2018-YB-04)
关键词
二代测序
血液透析
微生物多样性
Second generation sequencing technology
Hemodialysis
Microbial diversity
