The Feasibility of Metagenomic Next-Generation Sequencing to Identify Pathogens Causing Tuberculous Meningitis in Cerebrospinal Fluid

Frontiers in Microbiology

Suyue Pan¹  Yongjun Li²  Jian Zhang³  Jianzhao Zhang⁴  Guanghui Liu⁵  Shengnan Wang⁵  Xuan Li⁵  Yingwei Huang⁵  Dongmei Wang⁵  Yafang Hu⁵  Deqiang Zhao⁵  Haishan Jiang⁵  Yingli Chen⁵  Wei Li⁵  Yongming Wu⁵  Kaibin Huang⁵  Hongzhi Guan⁶  Huifang Xie⁷  Yanping Gong⁸  Ruixue Sun⁸  Xifang Nie⁸ 

[1] Peking Union Medical College, Beijing, China;BGI Genomics, BGI-Shenzhen, Shenzhen, China;Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou, China;Department of Neurology, Beijing Children’s Hospital, Capital Medical University, Beijing, China;Department of Neurology, Nanfang Hospital, Southern Medical University, Guangzhou, China;;Department of Neurology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences &Department of Neurology, Zhujiang Hospital, Southern Medical University, Guangzhou, China;Tianjin Medical Laboratory, BGI-Tianjin, BGI-Shenzhen, Tianjin, China;

关键词: cerebrospinal fluid;    Mycobacterium tuberculosis;    meningitis;    metagenomic next-generation sequencing;    early diagnosis;   

DOI  :  10.3389/fmicb.2019.01993

来源: DOAJ

【 摘 要 】

PurposeThe application of metagenomic next-generation sequencing (mNGS) in the diagnosis of tuberculous meningitis (TBM) remains poorly characterized. Here, we retrospectively analyzed data from patients with TBM who had taken both mNGS and conventional tests including culture of Mycobacterium tuberculosis (MTB), polymerase chain reaction (PCR) and acid-fast bacillus (AFB) stain, and the sensitivity and specificity of these methods were compared.MethodsWe retrospectively recruited TBM patients admitted to the hospital between December 2015 and October 2018. The first collection of cerebrospinal fluid (CSF) samples underwent both mNGS and conventional tests. In addition, patients with bacterial/cryptococcal meningitis or viral meningoencephalitis were mNGS positive controls, and a patient with auto-immune encephalitis was an mNGS negative control.ResultsTwenty three TBM patients were classified as 12 definite and 11 clinical diagnoses, which were based on clinical manifestations, pathogen evidence, CSF parameters, brain imaging, and treatment response. The mNGS method identified sequences of Mycobacterium tuberculosis complex (MBTC) from 18 samples (18/23, 78.26%). In patients with definite TBM, the sensitivity of mNGS, AFB, PCR, and culture to detect MTB in the first CSF samples were 66.67, 33.33, 25, and 8.33%, respectively. The specificity of each method was 100%. Among the four negative mNGS cases (4/23, 17.39%), three turned out positive by repeated AFB stain. The agreement of mNGS with the total of conventional methods was 44.44% (8/18). Combination of mNGS and conventional methods increased the detection rate to 95.65%. One patient was diagnosed as complex of TBM and cryptococcal meningitis, in which AFB stain and cryptococcal antigen enzyme immunoassay were positive and the DNA of Cryptococcus neoformans was detected by mNGS.ConclusionOur study indicates that mNGS is an alternative method to detect the presence of mycobacterial DNA in CSF samples from patients with TBM and deserves to be applied as a front-line CSF test.

【 授权许可】

Unknown