Molecular genetic test system for diagnosis of x-linked agammaglobulinemia

Abstract

Aim. Primary immunodeficiencies are disorders in which immune system is missing or does not function normally due to genetic disorders. X-linked agammaglobulinemia (XLA) is a rare genetic disorder in which don’t generate of mature B cells, which manifests as a complete or near-complete lack of proteins called gamma globulins, including antibodies, in their bloodstream. The BTK protein is the key regulator of β-cell maturation, so mutations in the BTK gene lead to hereditary agammaglobulinemia. Detection of mutations of the BTK gene is important for confirmation of the diagnosis in patients with XLA. Methods. DNA and RNA extraction, RT-PCR, direct Sanger sequencing of PCR fragments. Results. An analysis of blood samples from patients with XLA using a proposed test system was performed. It has been shown that the proposed test system allows for the initial screening of mutations in the BTK gene. Conclusions. The test system for the detection of mutations in patients with X-linked agammaglobulinemia, based on using of reverse transcriptase PCR and direct sequencing, was developed and tested on clinical samples. This system is proposed for the initial screening of BTK gene mutations inUkraine. Keywords: X-linked agammaglobulinemia, XLA, BTK, mutation, RT-PCR, primary immunodeficiency.