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Development of diagnosis method for LTBI with circulating miRNA

ABSTRACT: When exposed to M. tuberculosis, 25-50% become infected, of which 10-20% having several symptoms. Currently, there is no marker for judging the efficacy of M tuberculosis (Mtb) treatment, and it is judged based on the appearance of subjective symptoms and radiological findings. In this study, we attempted to establish a marker to distinguish patients who relapsed after TB treatment from patients who had recurrence after TB treatment using the information of circulating miRNA expression. Thirty two patients with tuberculosis infection aged 18 years or older were enrolled. 16 patients had onset or recurrence of TB and 16 patients with latent tuberculosis infection (LTBI) with positive test for Interferon gamma release assays (IGRA) and no chest X-ray examination. Total RNA from serum in exosome-rich fraction was extracted and then miRNA expression analysis was performed using a next-generation sequencer and NGS data analyzed using miRDeep2. Using expression information of nine miRNAs, TB and LTBI could be separated with 71.8% accuracy. The odds ratio also was 6.16, and the p-value was 3.20e-02, respectively. The expression of hsa-miR-122-5p was significantly different between TB and LTBI (p <0.05). Here we established a novel method to efficiently differentiate LTBI from patients with tuberculosis infection. Since this method can be performed in a low invasive manner such as blood sampling, it is promising tool for utilizing follow-up of TB treatment.

ORGANISM(S): Homo sapiens

PROVIDER: GSE149645 | GEO | 2021/03/18

REPOSITORIES: GEO

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Project description:Understanding the immune response to tuberculosis requires greater knowledge of humoral responses. To characterize antibody targets and the effect of disease parameters on target recognition, we developed a systems immunology approach that integrated detection of antibodies against the entire Mycobacterium tuberculosis proteome, bacterial metabolic and regulatory pathway information, and patient data. Probing ~4,000 M. tuberculosis proteins with sera from >500 suspected tuberculosis patients worldwide revealed that antibody responses recognized ~10% of the bacterial proteome. This result defines the immunoproteome of M. tuberculosis, which is rich in membrane-associated and extracellular proteins. Most serum reactivity during active tuberculosis focused onto ~0.5% of the proteome. Within this pool, which is selectively enriched for extracellular proteins (but not for membrane-associated proteins), relative target preference varied among patients. The shift in relative M. tuberculosis protein reactivity observed with active tuberculosis defines the evolution of the humoral immune response during M. tuberculosis infection and disease. Peripheral blood was collected from prospectively enrolled TB suspects among subjects seeking care for pulmonary symptoms at clinics associated with national TB control programs in 11 countries. M. tuberculosis proteome microarrays representing 4099 bacterial protein spots were probed with sera from 561 TB suspects. Based on the final diagnosis, they belonged to two classes: TB (n=254) and Non-TB Disease (n=307). In addition, healthy individuals negative to Latent TB Infection (LTBI neg, n=64) were also tested (negative control sera). Each serum was tested with a single array and no replicate experiments were performed. The reactivity of a serum to an M. tuberculosis protein (reactivity call) was defined based on the distribution of negative control sera intensity for that protein using Z-statistics. Based on the distribution of reactivity calls, 27 outlier samples reacting with more than 20 proteins were excluded from further analysis. The association of reactivity calls of each protein with TB/NTBD status of TB suspects was determined by estimating odds ratio and 95% confidence interval.

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Development of diagnosis method for LTBI with circulating miRNA

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