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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
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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
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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

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

    摘要
  • 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。

     

    Abstract:
      Objective  The aim of this study was to evaluate the potential diagnostic value of metagenomic next-generation sequencing(mNGS) for cryptococcus infection in the central nervous system(CNS).
      Methods  The clinical data of 6 patients with cryptococcal meningitis diagnosed with mNGS in Peking Union Medical College Hospital from January 2014 to December 2016 were retrospectively collected, including the routine examinations, the culture of cerebrospinal fluid (CSF), biochemical and cytologic tests, India Ink stain methods, and the sequencing results using BGISEQ-100 sequencing platform to identify the pathogenic microorganisms of CSF.
      Results  The 6 patients'median age was 51 years (range from 26 to 53 years), 4 males and 2 females. None of them had immunodeficiency disease. All had the headache and meningeal irritation, five with fever, and two with diplopia. Lumbar puncture revealed increased opening pressure, slightly increased pleocytosis and protein content, and normal/slightly decreased glucose in all cases. CSF India ink stain was positive in five patients. The antigen of cryptococcal meningitis was positive in 4 patients and the isolation by culture was positive in 2.Cryptococcus specific sequences were detected in all the six patients by mNGS, 5 patients infected with Cryptococcus neoformans and one with Cryptococcus gattii. The identified reads corresponding to Cryptococcus neoformans and Cryptococcus gattii ranged from 108 to 25 361 with genomic coverage rates from 0.19% to 29.00%.
      Conclusion  mNGS may be beneficial to make the diagnosis of cryptococcus infection in CNS, especially for the differential diagnosis of Cryptococcus gattii.

     

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