摘要
目的筛选棘球蚴病(包虫病)潜在诊断标志物的游离DNA(cell-free DNA,cfDNA)序列,并初步探讨其诊断价值。方法从西藏自治区采集确诊细粒棘球蚴病患者外周血,分离血浆,提取cfDNA并重测序;对测序数据进行生物信息学分析,并与人参考基因组和细粒棘球绦虫基因组综合比对,筛选诊断标志物候选序列;分别以细粒棘球蚴病患者和健康人血浆cfDNA为模板,采用qPCR分析诊断标志物候选序列的特异性和敏感性。结果共采集6份细粒棘球蚴病患者血浆,提取的血浆cfDNA条带较为均一,主要集中于150~200 bp;经重测序和序列比对获得一组候选序列,qPCR分析后初步筛选出一条101碱基的序列,用其检测8例细粒棘球蚴病患者血浆cfDNA,其中5例阳性,敏感性为62.5%;检测健康人血浆的特异性为87.5%(7/8)。结论细粒棘球蚴病患者血浆中存在虫源cfDNA,该序列具有作为诊断标志物的潜在价值。
Objective To screen cell-free DNA(cfDNA) sequences that can be used as a potential diagnostic marker of echinococcosis and to evaluate their sensitivity and specificity at diagnosing that condition. Methods cfDNA was extracted from the plasma of patients with echinococcosis from Tibet and subjected to genome resequencing. All sequence data were analyzed bioinformatically and mapped with the reference genomes of Echinococcus granulosus and humans to identify potential diagnostic markers. The sensitivity and specificity of potential diagnostic markers were evaluated using qPCR with cfDNA from patients and controls. Results cfDNA was extracted from the plasma of 6 patients with cystic echinococcosis. Results indicated that cfDNA was approximately 150-200 bp in length. All cfDNA samples were successfully resequenced. A number of candidate sequences were obtained by mapping the reference genomes of E. granulosus and humans and were compared to sequence data from the 6 patients. A 101-base sequence was obtained and evaluated using cfDNA from patients and controls. The sequence had a sensitivity of detecting cystic echinococcosis of 62.5%(5/8) and a specificity of 87.5%(7/8), indicating its potential use to diagnose cystic echinococcosis. Conclusion cfDNA from E. granulosus was found in plasma from patients with cystic echinococcosis, and cfDNA has potential value as a diagnostic marker.
作者
王莹
贡桑曲珍
庞华胜
伍卫平
张璟
沈玉娟
曹建平
WANG Ying;GONGSANG Quzhen;PANG Hua-sheng;WU Wei-ping;ZHANG Jing;SHENG Yu-juan;CAO Jian-ping(The National Institute of Parasitic Diseases,Chinese Center for Disease Control and Prevention,Chianese Center for Tropical Diseases Research,WHO Collaborating Centre for Tropical Diseases,National Center for International Research on Tropical Diseases,MOST Key Laboratory of Parasite and Vector Biology,MOH,Shanghai,China 200025;Tibet Center for Disease Control and Prevention,Key Laboratory of Kchino-coccosis Prevention and Control,National Health Commission,Lhasa,China 850000)
出处
《中国病原生物学杂志》
CSCD
北大核心
2020年第6期674-677,702,共5页
Journal of Pathogen Biology
基金
国家自然科学基金项目(No.81601792,81772224,8177225)
国家卫健委包虫病防治研究重点实验室开放课题。
