Isoelectrofocusing and PCR Amplification-Reverse Hybridization Assay in Evaluation of Alpha-1-Antitrypsin Deficiency

Anđelo Beletić,Nada Majkić-Singh,Ivana Obradović,Aleksandra Dudvarski-Ilić,Mirka Ilić,Dragica Radojković,Valentina Đorđević,Duško Mirković

doi:10.2478/v10011-009-0023-x

Abstract

Isoelectrofocusing and PCR Amplification-Reverse Hybridization Assay in Evaluation of Alpha-1-Antitrypsin DeficiencyAlpha-1-antitrypsin deficiency is a potentially lethal genetic disorder, which has pulmonary and liver manifestations. The standardized biochemical and molecular diagnostic protocol for detection of clinically relevant alleles is needed. The paper summarizes current concepts about AATD, describes the potentials of isoelectric focusing and PCR amplification-reverse allele specific oligonucleotide hybridization assay in the detection of affected individuals and shortly presents our experiences in the evaluation of AATD. We conclude that the systematic clinical laboratory approach to AATD might be based on the combination of mentioned methods, coordinated by alpha-1-antritrypsin quantification. Additionally, its complete medical implementation is achieved through teamwork between clinical chemists, molecular biologists and clinicians.

Highlights

Alpha-1-antitrypsin deficiency (AATD) is a potentially lethal genetic disorder, characterized by serum concentrations of alpha-1-antritrypsin (AAT) lower than 0.5 mg/L, which has liver and pulmonary manifestations [1, 2]
The paper summarizes current concepts about AATD, describes the potentials of isoelectric focusing and PCR amplification-reverse allele specific oligonucleotide hybridization assay in the detection of affected individuals and shortly presents our experiences in the evaluation of AATD
This paper summarizes the current concepts about AATD, describes the potentials of isoelectric focusing and the PCR amplification-reverse hybridization assay in the detection of affected individuals and shortly presents our experiences in the evaluation of AATD

Summary

Introduction

Alpha-1-antitrypsin deficiency (AATD) is a potentially lethal genetic disorder, characterized by serum concentrations of alpha-1-antritrypsin (AAT) lower than 0.5 mg/L, which has liver and pulmonary manifestations [1, 2]. Further polymerization leads to the formation of insoluble inclusions, which trigger ER stress and represent hallmarks of AAT liver disease [1, 4, 7, 12] This feature classifies Z-associated AATD as the paradigm of conformational diseases [1]. Are mostly associated with the homozygous presence of Z or Mmalton alleles or their mutual presence in heterozygous genotypes It is not common for the same individual to have both the hepatic and pulmonary disease due to the AATD [1, 2, 4]. A precisely balanced definition of the adult population to be screened is a prerequisite for successful screening, in order to avoid limitations seen both in large population-based screening and in small targeted detection programs in high risk groups [4, 17]

Laboratory methods for the evaluation of AATD

Findings

Conclusion