People living with human immunodeficiency virus (HIV) infections (PLHIV) have a 15- to 21-fold increased risk of developing tuberculosis (TB), and TB is the leading cause of morbidity and mortality among PLHIV. Novel approaches that allow early diagnosis and treatment monitoring of both HIV and TB are essential to improve patient outcomes. We developed and validated an immuno-affinity liquid chromatography–tandem mass spectrometry (ILM) assay that simultaneously quantifies single peptides derived from HIV-1 p24 and Mycobacterium tuberculosis (Mtb) 10-kDa culture filtrate protein (CFP10) in trypsin-digested serum derived from cryopreserved serum archives of cohorts of adults and children with/without HIV and TB. ILM p24 and CFP10 results demonstrated good intra-laboratory precision and accuracy, with recovery values of 96.7% to 104.6% and 88.2% to 111.0%, total within-laboratory precision (CV) values of 5.68% to 13.25% and 10.36% to 14.92%, and good linearity (r² > 0.99) from 1.0 to 256.0 pmol/L and 0.016 to 16.0 pmol/L, respectively. In cohorts of adults (n = 265) and children (n = 512) with HIV and/or TB, ILM detected p24 and CFP10 demonstrated 85.7% to 88.9% and 88.9% to 100.0% diagnostic sensitivity for HIV-1 and TB, with 100% specificity for both, and detected HIV-1 infection earlier than 3 commercial p24 antigen/antibody immunoassays. Finally, p24 and CFP10 values measured in longitudinal serum samples from children with HIV-1 and TB distinguished individuals who responded to TB treatment from those who failed to respond or were untreated, and who developed TB immune reconstitution inflammatory syndrome. Simultaneous ILM evaluation of p24 and CFP10 results may allow for early TB and HIV detection and provide valuable information on treatment response to facilitate integration of TB and HIV diagnosis and management.

Characteristics of the p24/CFP10 ILM assay.

Tony Hu

Appearances