Plasma Matrix Metalloproteinases Signature as Biomarkers for Pediatric Tuberculosis Diagnosis: A Prospective Case-Control Study

Abstract

Diagnosing tuberculosis (TB) in children presents significant challenges, necessitating the identification of reliable biomarkers for accurate diagnosis. In this study, we investigated plasma matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) as potential diagnostic markers. A prospective case-control study involved 167 children classified into confirmed TB, unconfirmed TB, and unlikely TB control groups. Plasma levels of MMPs (MMP 1, 2, 3, 7, 8, 9, 12, and 13) and TIMPs (TIMP 1, 2, 3, and 4) were measured using multiplex assays. Elevated baseline levels of MMP-1, MMP-2, MMP-7, MMP-9, TIMP-1, TIMP-2, TIMP-3, and TIMP-4 were observed in active TB cases compared to unlikely TB controls. Receiver operating characteristics (ROC) analysis identified MMP-1, MMP-2, MMP-9, and TIMP-1 as potential biomarkers with over 80% sensitivity and specificity. A three-MMP signature (MMP-1, MMP-2, and MMP-9) demonstrated 100% sensitivity and specificity. The findings suggest that a baseline MMP signature could serve as an accurate biomarker for diagnosing pediatric TB, enabling early intervention and effective management.

Keywords: MMPs; biomarkers; tuberculosis.

Figure 1

Elevated circulating levels of MMPs in children with active TB disease. The plasma levels of MMP 1, 2, 3, 7, 8, 9, 12, and 13 were measured in confirmed TB (n = 44), unconfirmed TB (n = 47), and unlikely TB (n = 76) individuals at baseline. The data are represented as scatter plots with each circle representing a single individual. p values were calculated using the Kruskal–Wallis test with Dunn’s post hoc for multiple comparisons.

Figure 2

Elevated circulating levels of TIMPs in children with active TB disease. The plasma levels of TIMP 1, 2, 3, and 4 were measured in confirmed TB (n = 44), unconfirmed TB (n = 47), and unlikely TB (n = 76) individuals at baseline. The data are represented as scatter plots with each circle representing a single individual. p values were calculated using the Kruskal–Wallis test with Dunn’s post hoc for multiple comparisons.

Figure 3

Plasma MMPs are associated with disease severity. The plasma levels of MMP 1, 3, 7, 8, and 13 were measured among the active TB children with minimal TB (n = 59) and severe TB (n = 32) individuals at baseline. The data are represented by scatter plots with each circle representing a single individual. p values were calculated using the Mann–Whitney U test.

Figure 4

ROC analysis to estimate the discriminatory power of MMP in children with active TB disease and unlikely TB. ROC analysis to estimate the sensitivity, specificity and AUC was performed using MMP 1, 2, 3, 7, 8, 9, 12, and 13 and TIMP 1, 2, 3, and 4 to estimate the capacity of these factors to distinguish individuals with (A) confirmed TB vs. unlikely TB and (B) unconfirmed TB vs. unlikely TB. ROC = receiver operator characteristics.

Figure 4

ROC analysis to estimate the discriminatory power of MMP in children with active TB disease and unlikely TB. ROC analysis to estimate the sensitivity, specificity and AUC was performed using MMP 1, 2, 3, 7, 8, 9, 12, and 13 and TIMP 1, 2, 3, and 4 to estimate the capacity of these factors to distinguish individuals with (A) confirmed TB vs. unlikely TB and (B) unconfirmed TB vs. unlikely TB. ROC = receiver operator characteristics.

Figure 5

Identification of biomarkers showing the strongest association using a combination of MMP biomarkers in active TB disease. CombiROC model analysis shows the chemokines with the highest accuracy in discriminating confirmed TB and unconfirmed TB disease from unlikely TB. ROC curves for comparing multiple markers and their combinations between confirmed TB and unconfirmed TB versus unlikely TB. (A) Confirmed TB vs. unlikely TB; (B) unconfirmed TB vs. unlikely TB are shown.