Lateral flow assay for simultaneous detection of cellular- and humoral immune responses

Clin Biochem. 2011 Oct;44(14-15):1241-6. doi: 10.1016/j.clinbiochem.2011.06.983. Epub 2011 Jul 6.

Abstract

Objective: The development of a cytokine detection assay suitable for detection of multiple biomarkers for improved diagnosis of mycobacterial diseases.

Design and methods: A lateral flow (LF) assay to detect IL-10 was developed utilizing the up-converting phosphor (UCP) reporter-technology. The assay was evaluated using blood samples of leprosy patients. Multiplex applications were explored targeting: 1) IL-10 and IFN-γ in assay buffer; 2) IL-10 and anti-phenolic glycolipid (PGL-I) antibodies in serum from leprosy patients.

Results: Detection of IL-10 below the targeted level of 100pg/mL in serum was shown. Comparison with ELISA showed a quantitative correlation with R(2) value of 0.92. Multiplexing of cytokines and simultaneous detection of cytokine and antibody was demonstrated.

Conclusions: The UCP-LF IL-10 assay is a user-friendly, rapid alternative for IL-10 ELISAs, suitable for multiplex detection of different cytokines and can be merged with antibody-detection assays to simultaneously detect cellular- and humoral immunity.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Bacterial / analysis
  • Biological Assay / methods*
  • Biomarkers / blood
  • Buffers
  • Cytokines / blood*
  • Cytokines / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Glycolipids / analysis
  • Humans
  • Immunity, Humoral
  • Interferon-gamma / analysis*
  • Interferon-gamma / metabolism
  • Interleukin-10 / blood*
  • Leprosy / blood
  • Leprosy / diagnosis*
  • Leprosy / immunology*
  • Mycobacterium Infections / diagnosis
  • Mycobacterium Infections / immunology
  • Reference Values
  • Sensitivity and Specificity

Substances

  • Antigens, Bacterial
  • Biomarkers
  • Buffers
  • Cytokines
  • Glycolipids
  • IL10 protein, human
  • phenolic glycolipid I, Mycobacterium leprae
  • Interleukin-10
  • Interferon-gamma