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隐球菌脑膜炎患者脑脊液宏基因组二代测序结果分析

葛瑛 范思远 陈健华 申奥 窦洪涛 肖盟 徐英春 宫燕萍 任海涛 吴红龙 关鸿志

葛瑛, 范思远, 陈健华, 申奥, 窦洪涛, 肖盟, 徐英春, 宫燕萍, 任海涛, 吴红龙, 关鸿志. 隐球菌脑膜炎患者脑脊液宏基因组二代测序结果分析[J]. 协和医学杂志, 2019, 10(6): 605-611. doi: 10.3969/j.issn.1674-9081.2019.06.010
引用本文: 葛瑛, 范思远, 陈健华, 申奥, 窦洪涛, 肖盟, 徐英春, 宫燕萍, 任海涛, 吴红龙, 关鸿志. 隐球菌脑膜炎患者脑脊液宏基因组二代测序结果分析[J]. 协和医学杂志, 2019, 10(6): 605-611. doi: 10.3969/j.issn.1674-9081.2019.06.010
Ying GE, Si-yuan FAN, Jian-hua CHEN, Ao SHEN, Hong-tao DOU, Meng XIAO, Ying-chun XU, Yan-ping GONG, Hai-tao REN, Hong-long WU, Hong-zhi GUAN. Analysis of Metagenomic Next-generation Sequencing of Cerebrospinal Fluid from Patients with Cryptococcal Meningitis[J]. Medical Journal of Peking Union Medical College Hospital, 2019, 10(6): 605-611. doi: 10.3969/j.issn.1674-9081.2019.06.010
Citation: Ying GE, Si-yuan FAN, Jian-hua CHEN, Ao SHEN, Hong-tao DOU, Meng XIAO, Ying-chun XU, Yan-ping GONG, Hai-tao REN, Hong-long WU, Hong-zhi GUAN. Analysis of Metagenomic Next-generation Sequencing of Cerebrospinal Fluid from Patients with Cryptococcal Meningitis[J]. Medical Journal of Peking Union Medical College Hospital, 2019, 10(6): 605-611. doi: 10.3969/j.issn.1674-9081.2019.06.010

隐球菌脑膜炎患者脑脊液宏基因组二代测序结果分析

doi: 10.3969/j.issn.1674-9081.2019.06.010
基金项目: 

国家重点研发计划罕见病队列研究项目 2016YFC0901501

宏基因组学快速感染诊断产品和传染病原分类方法开发 2018ZX10305409

详细信息
    通讯作者:

    关鸿志 电话:010-69156377, E-mail:guanhz@263.net

  • 中图分类号: R515;R446.5

Analysis of Metagenomic Next-generation Sequencing of Cerebrospinal Fluid from Patients with Cryptococcal Meningitis

More Information
    Corresponding author: GUAN Hong-zhi Tel: 86-10-69156377, E-mail:guanhz@263.net
  • 摘要:   目的  评价脑脊液宏基因组二代测序(metagenomic next-generation sequencing, mNGS)技术在隐球菌脑膜炎诊断中的潜在应用价值。  方法  回顾性收集北京协和医院2014年1月至2016年12月采用脑脊液mNGS技术辅助确诊的6例隐球菌脑膜炎患者临床资料, 包括脑脊液常规、生化、细胞学、培养、墨汁染色等实验室检测结果, 应用BGISEQ-100测序平台进行脑脊液病原测序, 综合分析患者的临床、实验室和辅助检查结果。  结果  6例患者中, 男性4例, 女性2例, 年龄26~53岁, 中位年龄51岁。6例患者均无免疫缺陷性疾病, 有头痛、脑膜刺激症状及颅内压增高表现, 5例患者出现发热, 2例患者出现复视。脑脊液常规检查示白细胞和蛋白轻度升高, 糖正常或轻度降低; 5例患者脑脊液墨汁染色阳性, 4例患者脑脊液隐球菌抗原检测阳性, 2例患者脑脊液真菌培养阳性。脑脊液mNGS检测隐球菌核酸序数为108~25 361, 基因覆盖率0.19%~29.00%;5例提示新型隐球菌感染, 其中3例经PCR检测证实为新型隐球菌感染; 1例提示格特隐球菌感染, 经生物质谱仪检测证实为格特隐球菌感染。  结论  脑脊液mNGS技术可准确判断隐球菌感染, 并对鉴别格特隐球菌具有一定优势, 有助于降低免疫功能正常人群隐球菌脑膜炎的漏诊率。
    利益冲突  无
  • 图  1  隐球菌脑膜炎患者的头颅磁共振成像

    左列为T1加权像, 右列为T2加权像, A、B.发病2个月时, 双侧基底节区显示多发囊性病变, C、D.发病12个月、治疗11个月后, 双侧基底节区多发囊性病灶范围扩大, E、F.抗真菌治疗8个月后, 双侧基底节区多发囊性病灶缩小

    图  2  脑脊液墨汁染色可见大量隐球菌(自然沉淀法侯氏沉淀器制片, ×200)

    图  3  格特隐球菌序列数高的隐球菌脑膜炎患者脑脊液宏基因二代测序检测结果

    A.核酸序列在格特隐球菌基因组的位置; B.序列分布图

    图  4  新型隐球菌序列数高的隐球菌脑膜炎患者脑脊液宏基因二代测序检测结果

    A.核酸序列在新型隐球菌基因组的位置; B.序列分布图

    图  5  隐球菌脑膜炎患者脑脊液pcr验证结果

    A.患者1(泳道064)和患者2(泳道219-2),N为空白对照,M为Trans2K DNA Marker; B.患者4(泳道434),N为空白对照,M为DL2000 DNA Marker

    表  1  6例隐球菌脑膜炎患者脑脊液实验和mNGS检测结果[5-6]

    编号 年龄(岁) 实验室检测 mNGS检测
    颅内压(mm H2O) 白细胞(×106/L) 蛋白(g/L) 糖(mmol/L) 氯化物(mmol/L) 墨汁染色 真菌培养 C-Ag 检测病原 核酸序列数 基因覆盖率(%)
    1 50 >330 176 0.84 3.40 119 + + 1:1024 格特 824 0.76
    2 26 >330 45 0.50 2.16 117 + - 1:1024 新型 824 0.45
    3 53 >330 13 0.63 2.48 120 + - 1:512 新型 25 361 29.00
    4 36 220 10 0.46 2.65 118 + - / 新型 8004 10.00
    5 42 260 28 0.86 1.74 121 - + 1:1024 新型 108 0.19
    6 49 >330 50 0.47 3.44 124 + - / 新型 2083 3.20
    ERAS:加速康复外科;VTE:静脉血栓栓塞症
    下载: 导出CSV
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  • 收稿日期:  2019-07-09
  • 刊出日期:  2019-11-30

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