ESAT-6 and CFP-10- induced cytokine response in FluoroSpot.

<p>ESAT-6 induced cytokine immune response in 200.000 PBMCs/well in participants with active tuberculosis (TB, circle, n = 18), past tuberculosis (past TB, inverted triangle, n = 10), latent infection with <i>M</i>. <i>tuberculosis</i> (LTBI, square, ESAT-6-induced n = 24, CFP-10-induced n = 22), EliSpot-negative individuals (control, triangle, ESAT-6-induced n = 17, CFP-10-induced n = 19) was analysed. Groups had been defined according to the combination of their ESAT-6 and CFP-10 induced IFN-γ EliSpot-IGRA test result and clinical data. The number of IL-2⁺ (A), INF-γ⁺ (B), IL-2⁺ INF-γ^- (C), IL-2^- INF-γ⁺ (D) and IL-2⁺ INF-γ⁺ (E) spot-forming cells (SFC) were enumerated by FluoroSpot. (F) Mean proportion of ESAT-6 (top row) and CFP-10 (bottom row) -specific cytokine secreting cells for individuals with tuberculosis, past tuberculosis and LTBI are depicted as pie charts (light grey = IL-2⁺ INF-γ^-, black = IL-2^- INF-γ⁺ and dark grey = IL-2⁺ INF-γ⁺ secreting cells). Mann-Whitney U-test for non-parametric data was used for comparative analysis. A p-value of <0.05 was considered significant.</p

Flow chart of patients included in this study.

<p>TB = tuberculosis; LTBI = latent infection with <i>M</i>. <i>tuberculosis;</i> control = healthy individual with negative EliSpot-IGRA result; E = ESAT-6; C = CFP-10; dark grey = positive test result in the EliSpot-IGRA; light grey = negative test result in the EliSpot-IGRA.</p

Demographic characteristics of study subjects by groups.

<p>TB = tuberculosis.</p><p>LTBI = latent infection with <i>M</i>. <i>tuberculosis</i>.</p><p>m = male.</p><p>f = female.</p><p>n = number of cases.</p><p>Demographic characteristics of study subjects by groups.</p

Detection of INF-γ⁺ in EliSpot-IGRA and their concordance with INF-γ⁺ results in FluoroSpot.

<p>ESAT-6 (A) and CFP-10 (B) -induced INF-γ⁺ immune response in 200.000 PBMCs/well in participants with active tuberculosis (TB, circle, n = 18), past tuberculosis (past TB, inverted triangle, n = 10), latent infection with M. tuberculosis (LTBI, square, ESAT-6-induced n = 24, CFP-10-induced = 22) and EliSpot-negative individuals (control, triangle, ESAT-6-induced n = 17, CFP-10-induced n = 19) was analysed. LTBI and controls had been defined according to their ESAT-6 and CFP-10-induced IFN-γ EliSpot-IGRA test result and clinical data. Number of INF-γ⁺ spot-forming cells (SFC) was enumerated by EliSpot. ESAT-6 (C) and CFP-10 (D) induced- INF-γ⁺ SFC in EliSpot-IGRA (solid symbols) and FluoroSpot (open symbols) were analysed as matched pairs (connected with lines), differences were calculated using Wilcoxon signed rank test. Correlation between the number of ESAT-6 (E) and CFP-10 (F) specific INF-γ⁺ spot-forming cells (SFC) in PBMC of 69 donors detected by FluoroSpot and EliSpot-IGRA. Concordance between EliSpot-IGRA and FluoroSpot results were assessed using R² coefficient. Agreement by Bland–Altman test was expressed as mean difference (horizontal solid line) and 95% limits of agreement (dashed line) between ESAT-6 (G) and CFP-10 (H) induced- INF-γ⁺ SFC in FluoroSpot compared to EliSpot-IGRA.</p

Additional file 2: of Shedding light on the performance of a pyrosequencing assay for drug-resistant tuberculosis diagnosis

Pyrosequencing (PSQ) Performance Dataset. (TXT 400 kb

Additional file 1: Table S1. of Shedding light on the performance of a pyrosequencing assay for drug-resistant tuberculosis diagnosis

Pyrosequencing (PSQ) Success of Each Gene Target by Smear- and Culture-Status. Table S2. Proportion of Successful Pyrosequencing (PSQ) Reactions by Smear- and Culture-Status. (DOCX 18 kb

Cumulative incidence of grade 4–5 SAEs under 6 months BPaLM and standard of care.

The mean cumulative incidence of grade 4–5 SAEs ever experienced to each of 12 anti-TB drugs is shown for Strategy (1) (6 months BPaLM, DST upfront, repeat DST every 4 months, BPaLC if Mfx stopped) as compared to Strategy (7) (standard of care 9- to 18-month regimens based on results of upfront DST, repeat DST every 4 months). Estimates are provided per individual, averaged over the entire cohort initiating treatment. The mean estimate is shown by the bar, with 95% UIs represented as error bars. BPaLM, bedaquiline, pretomanid, linezolid, moxifloxacin; SAE, grade 4–5 severe adverse event; TB, tuberculosis; UI, uncertainty interval.</p

Cohort prevalence of <i>M. tb</i>. resistance to key drugs at treatment initiation.

The proportion of the cohort with primary resistance to each drug is plotted, as described by M. tuberculosis whole genomic sequencing data from Moldova [30,35]. All those observations with rifampicin susceptibility were excluded, as per S2 Fig. *There was no resistance data for pretomanid; resistance was assumed to be at the same level as for delamanid. (PDF)</p

Markov state-transition diagram.

Transitions between states can occur as shown by the arrows. Though not receiving treatment, individuals in the “Active TB, no longer receiving treatment” state are subject to a low rate of self-cure, and so may still transition to the “Cured post-treatment” state. Asterisks (*) highlight the major mechanisms through which the choice of treatment intervention affects outcomes. LTFU, lost to follow-up; TB, tuberculosis. Images within this figure were obtained as icons from Microsoft with no license or terms of use: https://support.microsoft.com/en-us/office/insert-icons-in-microsoft-365-e2459f17-3996-4795-996e-b9a13486fa79?ui=en-us&rs=en-us&ad=us. (PDF)</p

Life years achieved under each RR-TB treatment strategy.

BPaL, bedaquiline, pretomanid, linezolid; BPaLC, bedaquiline, pretomanid, linezolid, clofazimine; BPaLM, bedaquiline, pretomanid, linezolid, moxifloxacin; UI, uncertainty interval. Strategies are listed in the same order as Table 3. Mean values are shown with accompanying 95% UIs in parentheses. (PDF)</p