A Novel LC-MS/MS-Based Method for the Diagnosis of ADA2 Deficiency from Dried Plasma Spot

Molecules. 2021 Sep 21;26(18):5707. doi: 10.3390/molecules26185707.

Abstract

Adenosine Deaminase 2 Deficiency (DADA2) (OMIM: 607575) is a monogenic, autoinflammatory disease caused by the loss of functional homozygous or heterozygous mutations in the ADA 2 gene (previously CECR1, Cat Eye Syndrome Chromosome Region 1). A timely diagnosis is crucial to start Anti-TNF therapies that are efficacious in controlling the disease. The confirmation of DADA2 is based on DNA sequencing and enzymatic assay. It is, thus, very important to have robust and reliable assays that can be rapidly utilized in specialized laboratories that can centralize samples from other centers. In this paper, we show a novel enzymatic assay based on liquid chromatography-tandem mass spectrometry that allows the accurate determination of the ADA2 enzyme activity starting from very small amounts of plasma spotted on filter paper (dried plasma spot). The method allows significantly distinguishing healthy controls from affected patients and carriers and could be of help in implementing the diagnostic workflow of DADA2.

Keywords: Adenosine Deaminase; DADA2; LC-MS/MS; autoinflammatory disease; enzyme deficiency.

MeSH terms

  • Adenosine Deaminase / blood*
  • Adenosine Deaminase / genetics
  • Adenosine Deaminase / metabolism
  • Adolescent
  • Adult
  • Agammaglobulinemia / diagnosis*
  • Biomarkers / blood*
  • Child
  • Dried Blood Spot Testing
  • Female
  • Homozygote
  • Humans
  • Male
  • Middle Aged
  • Mutation
  • Phenotype
  • Severe Combined Immunodeficiency / diagnosis*
  • Tandem Mass Spectrometry
  • Tumor Necrosis Factor Inhibitors / metabolism

Substances

  • Biomarkers
  • Tumor Necrosis Factor Inhibitors
  • Adenosine Deaminase

Supplementary concepts

  • Severe combined immunodeficiency due to adenosine deaminase deficiency