Two Small Extracellular Vesicle sRNAs Derived From Mycobacterium tuberculosis Serve as Diagnostic Biomarkers for Active Pulmonary Tuberculosis

Abstract

The rapid diagnosis of tuberculosis (TB) is of great significance for the control and treatment of TB. However, TB remains a major healthy, social, and economic burden worldwide because of the lack of ideal diagnostic biomarkers. Mycobacterium tuberculosis (M. tuberculosis)-encoded small RNA (sRNA) is a class of regulation small RNA. Several studies have identified M. tuberculosis encoded-sRNAs in the serum/plasm of M. tuberculosis-infected patients. Small extracellular vesicles are small membrane vesicles secreted by many cell types during physiological and pathological conditions. Recent evidence has indicated that most of the nucleic acids in the serum/plasma are packaged in the small extracellular vesicles and could serve as ideal diagnostic biomarkers. In this study, we attempted a novel approach for TB diagnosis: targeting small extracellular vesicles M. tuberculosis encoded sRNA (sRNA) by qRT-PCR. The results showed that M. tuberculosis-encoded ASdes and MTB-miR5 only existed in tuberculosis patients and have the potential to serve as a sensitive and accurate methodology for TB diagnosis.

Identification of RNU44 as an Endogenous Reference Gene for Normalizing Cell-Free RNA in Tuberculosis

“…Emerging studies have demonstrated the potential diagnostic value of cf-RNAs for TB, including host- and pathogen-derived sRNAs [ 3 , 25 , 26 ]. However, many technical and sample-related factors may impact the results of cf-RNA quantification, such as specimen type, storage condition, RNA isolation method, and the donor's physical state [ 4 ].…”

Bacterial extracellular vesicles: biotechnological perspective for enhanced productivity

“…Reduction in infections severity and repair of the intestinal epithelium promoted by KGF-2 Carvalho et al ( 2019 ) Antimicrobial Antimicrobial treatments Burkholderia thailandensis 4-hydroxy-3-methyl-2-(2-non-enyl)-quinoline and long chain rhamnolipids Biofilm Streptococcus mutans (Oral pathogen) Biofilm integrity reduction Reduction of S. mutans viability Yihui Wang et al ( 2021a , b ) Protection of stored foods and increase of shelf life Lactobacillus plantarum Molecules derived from L. plantarum Shewanella putrefaciens (tuna spoilage cause) Inhibition of Shewanella putrefaciens growth. Inhibition of oxidation reactions in tuna stored for five days at 4 °C Lee et al ( 2021 ) Biomarkers Tuberculosis diagnosis Mycobacterium tuberculosis Small RNAs: ASdes and MTB-miR5 – qRT-PRC detection of ASdes and MTB-miR5 in vesicles from tuberculosis-infected patients Lu et al ( 2021 ) Other biotechnology areas Study of membrane proteins Escherichia coli OMPs: OmpG, FhuA, Tsx, BamA – OMVs maintain the unique amphipathic environment provided by lipid bilayers. The proteins under study retained their orientation, native structure, and functions Thoma et al ( 2018 ) Study of phospholipid permeability vesicle based Escherichia coli BL21 DE3 Porins, LPS, other cargo molecules – Allows estimation of cell wall permeability of Gram-negative bacteria facilitated by porins.…”

“…For example, L. plantarum MVs have been found to inhibit the growth of Shewanella putrefaciens in tuna (Lee et al 2021 ), and Helicobacter pylori OMVs coated with nanoparticles of poly(lactic-co-glycolic acid), prevents the adhesion of H. pylori to epithelial cells (Zhang et al 2019a , b ). Tuberculosis could be diagnosed by detecting specific sRNAs, such as Asdes and MTB-miR5, encapsulated in M. tuberculosis MVs that could be obtained from blood samples of infected patients (Lu et al 2021 ). Furthermore, BEVs can be used to support enzymes that degrade contaminating compounds (Alves et al 2015 ; Thoma et al 2018 ).…”

Small RNAs Asserting Big Roles in Mycobacteria

“…Later work revealed that ASdes, ASpks, AS1726, and AS1890 were also present in Bacillus Calmette Guerin (BCG) mycobacterial cultures [22]. ASdes was subsequently detected in the plasma of 15 of 27 TB patients and 6 of 24 BCG vaccinated individuals [22][23][24]. Additional RNA sequencing approaches have since broadened the number of small RNAs detected in exponential phase Mtb cultures, with 19 antisense and 20 intergenic sRNAs identified [10].…”

“…Later work revealed that ASdes, ASpks, AS1726, and AS1890 were also present in Bacillus Calmette Guerin (BCG) mycobacterial cultures [ 22 ]. ASdes was subsequently detected in the plasma of 15 of 27 TB patients and 6 of 24 BCG vaccinated individuals [ 22 , 23 , 24 ].…”