BACKGROUND: Laboratory evaluation of alpha 1-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.
BACKGROUND: Laboratory evaluation of alpha 1-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.
Authors: David A Bergin; Emer P Reeves; Paula Meleady; Michael Henry; Oliver J McElvaney; Tomás P Carroll; Claire Condron; Sanjay H Chotirmall; Martin Clynes; Shane J O'Neill; Noel G McElvaney Journal: J Clin Invest Date: 2010-11-08 Impact factor: 14.808
Authors: Kevin Molloy; Craig P Hersh; Valerie B Morris; Tomás P Carroll; Catherine A O'Connor; Jessica A Lasky-Su; Catherine M Greene; Shane J O'Neill; Edwin K Silverman; Noel G McElvaney Journal: Am J Respir Crit Care Med Date: 2014-02-15 Impact factor: 21.405
Authors: Alessandro N Franciosi; Brian D Hobbs; Oliver J McElvaney; Kevin Molloy; Craig Hersh; Louise Clarke; Cedric Gunaratnam; Edwin K Silverman; Tomás P Carroll; Noel G McElvaney Journal: Am J Respir Crit Care Med Date: 2020-07-01 Impact factor: 21.405
Authors: Tomás P Carroll; Catherine A O'Connor; Olwen Floyd; Joseph McPartlin; Dermot P Kelleher; Geraldine O'Brien; Borislav D Dimitrov; Valerie B Morris; Clifford C Taggart; Noel G McElvaney Journal: Respir Res Date: 2011-07-13
Authors: Irene Belmonte; Miriam Barrecheguren; Rosa M López-Martínez; Cristina Esquinas; Esther Rodríguez; Marc Miravitlles; Francisco Rodríguez-Frías Journal: Int J Chron Obstruct Pulmon Dis Date: 2016-10-11