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Diagnosis of paediatric tuberculosis by optically detecting two virulence factors on extracellular vesicles in blood

Wenshu Zheng1,2, Sylvia M LaCourse3,4, Bofan Song5

  • 1Center for Cellular and Molecular Diagnostics, Tulane University School of Medicine, New Orleans, LA, USA.

Nature Biomedical Engineering
|August 19, 2022
PubMed

Insights

A new nanoparticle-enhanced immunoassay detects tuberculosis (TB) virulence factors in blood extracellular vesicles. This sensitive assay shows promise for diagnosing pediatric TB, even in missed or undiagnosed cases, especially in children with HIV.

Area of Science:

  • Immunology and Infectious Diseases
  • Nanotechnology in Diagnostics
  • Point-of-Care Testing

Background:

  • Tuberculosis (TB) diagnosis, especially in children, is challenging, leading to significant mortality from missed or delayed diagnoses.
  • Current microbiological assays lack sensitivity and specificity, particularly in severely immunocompromised populations like children living with HIV.
  • Need for rapid, sensitive, and specific blood-based diagnostic tools for both pulmonary and extrapulmonary tuberculosis.

Purpose of the Study:

  • To develop and evaluate a novel nanoparticle-enhanced immunoassay for detecting Mycobacterium tuberculosis virulence factors.
  • To assess the assay's performance in diagnosing pediatric tuberculosis in hospitalized children living with HIV.
  • To explore the potential for a portable, smartphone-operable version for point-of-care use in resource-limited settings.

Main Methods:

  • Development of a nanoparticle-enhanced immunoassay detecting lipoarabinomannan (LAM) and its carrier protein on extracellular vesicles.
  • Assay read-out using dark-field microscopy.
  • Validation in a cohort of 147 hospitalized, severely immunosuppressed children living with HIV, comparing results with microbiological assays and undiagnosed cases.

Main Results:

  • The assay detected 74% of pediatric tuberculosis cases (58/78) in the study cohort.
  • It identified 73% (48/66) of cases missed by conventional microbiological assays.
  • The assay also diagnosed 80% (8/10) of cases that were undiagnosed during the study period and distinguished TB from latent TB infection in non-human primates.

Conclusions:

  • The nanoparticle-enhanced immunoassay demonstrates high sensitivity and specificity for detecting tuberculosis in a vulnerable pediatric population.
  • The assay's ability to identify missed and undiagnosed cases highlights its potential to improve TB diagnostics.
  • Adaptation for smartphone operation suggests feasibility for point-of-care application, particularly in resource-limited settings.

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