Validation of an immunoassay to measure plasminogen-activator inhibitor-1 concentrations in human saliva

Biochem Med (Zagreb). 2014;24(2):258-65. doi: 10.11613/BM.2014.028. Epub 2014 Jun 15.

Abstract

Introduction: We have previously shown that the concentrations of D-dimer are significantly elevated in saliva compared with plasma. Saliva offers several advantages compared with blood analysis. We hypothesised that human saliva contains plasminogen activator inhibitor-1 (PAI-1) and that the concentrations are not affected by the time of saliva collection. The aim was to adopt and validate an immunoassay to quantify PAI-1 concentrations in saliva and to determine whether saliva collection time has an influence in the measurement.

Materials and methods: Two saliva samples (morning and afternoon) from the same day were collected from healthy subjects (N = 40) who have had no underlying heart conditions. A customized AlphaLISA® immunoassay (PerkinElmer®, MA, USA) was adopted and used to quantify PAI-1 concentrations. We validated the analytical performance of the customized immunoassay by calculating recovery of known amount of analyte spiked in saliva.

Results: The recovery (95.03%), intra- (8.59%) and inter-assay (7.52%) variations were within the acceptable ranges. The median salivary PAI-1 concentrations were 394 pg/mL (interquartile ranges (IQR) 243.4-833.1 pg/mL) in the morning and 376 (129.1-615.4) pg/mL in the afternoon and the plasma concentration was 59,000 (24,000-110,000) pg/mL. Salivary PAI-1 did not correlate with plasma (P = 0.812).

Conclusion: The adopted immunoassay produced acceptable assay sensitivity and specificity. The data demonstrated that saliva contains PAI-1 and that its concentration is not affected by the time of saliva collection. There is no correlation between salivary and plasma PAI-1 concentrations. Further studies are required to demonstrate the utility of salivary PAI-1 in CVD risk factor studies.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Adult
  • Female
  • Healthy Volunteers
  • Humans
  • Immunoassay / standards*
  • Male
  • Plasminogen Activator Inhibitor 1 / analysis*
  • Reagent Kits, Diagnostic
  • Reproducibility of Results
  • Saliva / chemistry*
  • Sensitivity and Specificity
  • Time Factors

Substances

  • Plasminogen Activator Inhibitor 1
  • Reagent Kits, Diagnostic
  • SERPINE1 protein, human