Issue 31, 2024

Microfluidic platform for the enzymatic pretreatment of human serum for the detection of the tuberculosis biomarker mannose-capped lipoarabinomannan

Abstract

Tuberculosis (TB) represents a major public health threat, with millions of new cases reported worldwide each year. A major hurdle to curtailing the spread of this disease is the need for low-cost, point-of-care (PoC) diagnostics. Mannose-capped lipoarabinomannan, a significant component of the Mycobacterium tuberculosis bacillus, has been heavily studied as a biomarker for TB, but with little success due to its complexation with endogenous components of body fluids in a manner that sterically interferes with its detection by ELISA and other immunoassays. Recent work by our group and others has shown that complexation can be disrupted with protein-denaturing protocols. By way of followup, we recently described an enzymatic digestion (Proteinase K) sample pretreatment that enables quantitative recovery of ManLAM spiked into healthy human control serum. Herein, we report on the transfer of our benchtop sample pretreatment methodology to an automated microfluidic platform. We show that this platform can be configured to: (1) carry out the pretreatment process with very little user interaction and, (2) yield recoveries for ManLAm spiked into control serum which are statistically indistinguishable from those achieved by the benchtop process. Plans to integrate this device with a portable sample reader as a possible basis for a PoC TB diagnostic system and analyze patient samples are briefly discussed.

Graphical abstract: Microfluidic platform for the enzymatic pretreatment of human serum for the detection of the tuberculosis biomarker mannose-capped lipoarabinomannan

Supplementary files

Article information

Article type
Technical Note
Submitted
25 אפר 2024
Accepted
11 יול 2024
First published
17 יול 2024

Anal. Methods, 2024,16, 5475-5481

Microfluidic platform for the enzymatic pretreatment of human serum for the detection of the tuberculosis biomarker mannose-capped lipoarabinomannan

C. J. Lambert, E. Clarke, D. Patel, L. B. Laurentius, B. K. Gale, H. J. Sant and M. D. Porter, Anal. Methods, 2024, 16, 5475 DOI: 10.1039/D4AY00772G

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