Evaluation of a new Sebia isoelectrofocusing kit for α1-antitrypsin phenotyping with the Hydrasys® System
F Zerimech, G Hennache, F Bellon… - Clinical Chemical …, 2008 - degruyter.com
F Zerimech, G Hennache, F Bellon, G Barouh, J Jacques Lafitte, N Porchet, M Balduyck
Clinical Chemical Laboratory Medicine, 2008•degruyter.comBackground: Laboratory evaluation of α1-antitrypsin (A1AT) deficiency is generally
performed by determination of A1AT concentrations and identification of specific allelic
variants by phenotyping. For this purpose, we evaluated a new Hydragel 18 A1AT
Isofocusing® kit on the semi-automatic Hydrasys® System (Sebia) for the determination of
A1AT phenotypes by isoelectrofocusing on ready-to-use agarose gels with specific
immunological detection. Methods: Serum samples from 66 patients were analysed with this …
performed by determination of A1AT concentrations and identification of specific allelic
variants by phenotyping. For this purpose, we evaluated a new Hydragel 18 A1AT
Isofocusing® kit on the semi-automatic Hydrasys® System (Sebia) for the determination of
A1AT phenotypes by isoelectrofocusing on ready-to-use agarose gels with specific
immunological detection. Methods: Serum samples from 66 patients were analysed with this …
Abstract
Background: Laboratory evaluation of α1-antitrypsin (A1AT) deficiency is generally performed by determination of A1AT concentrations and identification of specific allelic variants by phenotyping. For this purpose, we evaluated a new Hydragel 18 A1AT Isofocusing® kit on the semi-automatic Hydrasys® System (Sebia) for the determination of A1AT phenotypes by isoelectrofocusing on ready-to-use agarose gels with specific immunological detection.
Methods: Serum samples from 66 patients were analysed with this new kit in comparison with the conventional and manually performed isoelectrofocusing method on polyacrylamide gels with Coomassie Blue staining.
Results: A1AT phenotypes showed comparable iso-electrofocusing patterns in both systems. The good within-gel reproducibility of this kit was demonstrated using two normal serum samples (M1 and M1M2 phenotypes) and six pathological serum samples with different phenotypes (MS, SS, SZ, MZ, ZZ). A sensitivity study was undertaken by performing serial dilutions on a serum with a ZZ phenotype containing 0.27 g/L A1AT. The detection limit was 0.050 g/L.
Conclusions: This new method is highly specific, rapid and simple to perform. It improves identification of not only the most common but also various rare A1AT phenotypes. It appears to be suitable for routine analysis and screening applications in a clinical laboratory setting.
Clin Chem Lab Med 2008;46:260–3.
De Gruyter