Identification Of The Microbiome Present In Sputum Samples From Patients With Clinical Tuberculosis By NGS (Next Generation Sequencing)

Abstract

Tuberculosis is a reemerging disease that continues to be a public health problem worldwide with a high morbimortality rate. Infection with M. tuberculosis causes dysbiosis of the microbiota, condition that not only influences the latency of M.tuberculosisand the manifestation of the disease but also would be determinants of a greater progression of the disease and the failure of the recovery process. Another aspect to consider are the diagnostic methods which are varied and with limitations, however, thanks to the remarkable technological progress such as the New Generation Sequencing (NGS),there is currently access to the study of the microbiome that accompanies the human being in health conditions and associated with diseases such as tuberculosis. In the present study, the identification of the microbiome of samples from patients diagnosed with tuberculosis was performed, where 28 sputum samples were collected with clinical diagnosis of tuberculosis corresponding to: 20 without treatment, 6 with treatment, 2 Multidrogo - Resistant and 8 samples from patients without tuberculosis (control). For the identification of M. tuberculosis,TB1, TB2 and TB3primers were used. The molecular identification of the microbiome was carried out by Illumina MiSeq sequencing, using specific primers that target the V4 region of the 16S rRNA gene.According to the abundance of each study group, the following Phyla were presented: patients without tuberculosis (control) Firmicutes, Proteobacteria, Fusobacteria and Actinobacteria; patients with diagnosis of tuberculosis without treatment Proteobacteria, Bacteroidetes, Firmicutes and Actinobacteria; patients with diagnosis of tuberculosis who received treatment Firmicutes, Proteobacteria, Bacteroidetes and Proteobacteria; and patients with diagnosis of tuberculosis Multidrogo-resistant phyla Proteobacteria, Actinobacteria,
Bacteroidetes, Firmicutes and Actinobacteria.The study identified the diversity and abundance at phylum and genus level by Next Generation Sequencing in individuals diagnosed with tuberculosis in untreated, treated and multidrug-resistant patients, finding a marked variation in the microbiome present in sputum samples associated with tuberculosis.