重庆地区健康献血人群血液宏基因组分析与新发病原体鉴定  

Metagenomics analysis of healthy blood donors in Chongqing,with identification of emerging infectious diseases

作　　者：徐敏[1] 毛伟[2] 何涛[2] 卫昱燕 高瞻[1] 张玉[1] 张春红[2] 廖红梅[2] 刘鱼[1] 曾沛斌[1] 王憬惺[1] 何苗[1] 

机构地区：[1]中国医学科学院北京协和医学院输血研究所,四川成都610052 [2]重庆市血液中心

出　　处：《中国输血杂志》2016年第9期895-898,共4页Chinese Journal of Blood Transfusion

摘　　要：目的了解重庆地区健康无偿献血人群的微生物组,并调查可能存在的新发/再发病原体的流行情况,本研究利用深度测序的方法对重庆地区5 000人份健康无偿献血者标本进行了宏基因组学分析,以鉴定其中的新发/再发病原体。方法提取5 000人份血浆核酸总DNA,经随机引物扩增后,建库上机深度测序,用本课题组开发的Kraken Py软件分析宏基因组数据,并确定其中的微生物组,鉴定可能存在的新发/再发病原体。结果通过深度测序将获得的1.23 Gb的数据,其中包括2 146 844个有效读长,去人类DNA背景后,结果显示属于细菌的片段47条,属于病毒的片段21条,属于寄生虫的片段333条。最主要的病原体是刚地弓形虫(Toxoplasma gondii),其次是欧猥迭宫绦虫(Spirometra erinaceieuropaei),仅发现少量指环病毒科(Anelloviridae)的病毒成员如扭转病毒Torque teno virus等。另外,我们还发现在欧美发达国家已经被认为是能够造成输血安全威胁的病原体DNA片段,如疟原虫(Plasmodium spp.),婴儿利什曼原虫(Leishmania infantum)等。结论本研究结果显示了重庆地区健康无偿献血人群的微生物组结构,同时也揭示了这些健康献血者有可能携带的新发/再发病原体,提醒采供血系统工作者应结合当地新发传染病流行情况,合理的制定疫区或是传染病流行季节的筛查模式和献血者招募策略。另外,对于这些潜在风险也不必过于惊慌,因为这仅是微生物基因组片段结果,不代表该病原体仍具有感染性。Objective To investigate the microbiome in healthy blood donors in Chongqing to reveal some potential emerging and re-emerging infectious diseases. Methods Deep sequencing strategy was used to analyze the metagenomics from pooled sample collected from 5 000 volunteer blood donors from Chongqing. Genomic DNA was extracted and amplified with random primers PCR in order to construct a 250PE library which would be sent to conduct deep sequencing using IUumina Miseq. KrakenPy was used to analyze those assembled reads to illustrate the composition of microbiome as well as to identify the potential existed pathogens. Results A total of 1.23Gb raw data with 2 146 844 assembled reads were generated. After cleaning the human background, 50 reads from bacteria, 21 reads from viruses, and 333 reads from parasites were i- dentified. The major pathogens were shown as Toxoplasma gondii and Spirometra erinaceieuropaei. Only three kind of viruses from Anelloviridae were found including Torque teno virus, Torque teno midi virus, and Torque teno mini virus. Besides, some pathogens which were already considered as threats to blood safety in some developed countries were also discovered, such as Plasmodium spp. and Leishmania infantum. Conclusion Our investigation has revealed the metagenomics structure of healthy donors in Chongqing. The results showed us a though-provoking discovery of DNA fragments of some pathogens which might threaten blood safety. The imminent results allows us to think about regulation of reasonable screening methods as well as donor recruitment strategy in certain epidemic areas or seasons to ensure blood safety. However, on the contrary, we should not take the results too seriously because the DNA fragments could not represent the existence of infectious pathogens in the blood from the healthy donors in Chongqing.

关 键 词：新发病原体 献血人群 宏基因组学 高通量测序 血液安全 

分 类 号：R457.11[医药卫生—治疗学] R392.2[医药卫生—临床医学]