Abstract:

Objective: To evaluate the diagnostic performance of differentially expressed miRNA identified by gene chip in discriminating tuberculous meningitis (TBM) from viral meningitis (VM). Methods: A total of 28 TBM patients and 27 VM patients who were treated in Beijing Chest Hospital and Beijing Tiantan Hospital affiliated to Capital Medical University from Dec 2017 to Sep 2019 were recruited for validation using Real-time Quantitative PCR (qRT-PCR). Of 8 TBM patients and 5 VM patients who were recruited earlier were selected for genome-wide miRNA analysis by gene chip. The top 10 miRNA hub genes were screened by predicting the target genes of differentially expressed miRNAs, gene ontology (GO) enrichment analysis, Kyoto encyclopedia of genes and genomes (KEGG) signaling pathway analysis, and protein-protein interaction (PPI) network. qRT-PCR was used for further validation of the expression level of hsa-miR-21-5p between two groups, and the diagnostic value of miRNA was analyzed by area under receiver operating characteristic curve (AUC). Results: Totally 26 differentially expressed miRNAs (14 were up-regulated and 12 were down-regulated) were identified by miRNA microarray analysis, and further 3217 corresponding target genes were screened out by miRTarBase database. Through GO enrichment analysis and KEGG signal pathway analysis, 190 related target genes were screened out. The top 10 target genes as core target genes were defined by PPI. qRT-PCR verification of the relative expression of hsa-miR-21-5p based on gene chip samples showed that the expression level of has-miR-21-5p in TBM patients was significantly up-regulated than that in VM patients (15.890 (3.423, 25.581) vs. 0.807(0.614,0.955); Z=-2.355, P=0.019). The fold change (TBM/VM) in gene chip and qRT-PCR were 4.817 and 4.660, respectively, which was consistent between these two techniques. qRT-PCR analysis of hsa-miR-21-5p was further performed in 28 TBM patients and 27 VM patients. The results showed that the relative expression level of hsa-miR-21-5p in TBM patients was significantly higher than that in VM patients (4.825 (2.433, 21.362) vs. 0.204 (0.112, 0.711); Z=-5.000, P=0.000). The AUC for distinguishing TBM patients from VM patients was 0.893 (0.806-0.980), with a sensitivity of 85.7% and a specificity of 88.9%. Conclusion: As a significantly differentially expressed miRNA in gene chip analysis, has-miR-21-5p is significantly up-regulated in patients with TBM than VM patients, which can be used as a potential biomarker to differentiate TBM from VM.

Key words: Meningitis,bacterial, Meningitic,viral, Diagnosis, Microchip analytical procedures, MicroRNAs