Zerimech Farid, Hennache Genevieve, Bellon Franck, Barouh Guy, Jacques Lafitte Jean, Porchet Nicole, Balduyck Malika
Laboratoire de Biochimie Biologie Moléculaire, Centre Hospitalier Régional et Universitaire de Lille, Lille, France.
Clin Chem Lab Med. 2008;46(2):260-3. doi: 10.1515/CCLM.2008.036.
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.
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.
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.
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.
α1 -抗胰蛋白酶（A1AT）缺乏症的实验室评估通常通过测定A1AT浓度以及通过表型分析鉴定特定等位基因变体来进行。为此，我们在半自动Hydrasys系统（Sebia）上评估了一种新型Hydragel 18 A1AT等聚焦试剂盒，该试剂盒通过在即用型琼脂糖凝胶上进行等电聚焦并采用特定免疫检测法来测定A1AT表型。
使用该新型试剂盒对66例患者的血清样本进行分析，并与在聚丙烯酰胺凝胶上进行考马斯亮蓝染色的传统手动等电聚焦方法进行比较。
两种系统中A1AT表型均显示出可比的等电聚焦模式。使用两份正常血清样本（M1和M1M2表型）以及六份具有不同表型的病理血清样本（MS、SS、SZ、MZ、ZZ）证明了该试剂盒良好的凝胶内重复性。通过对一份含有0.27 g/L A1AT的ZZ表型血清进行系列稀释开展了一项灵敏度研究。检测限为0.050 g/L。
这种新方法具有高度特异性，操作快速且简单。它不仅能更好地鉴定最常见的A1AT表型，还能鉴定各种罕见的A1AT表型。它似乎适用于临床实验室环境中的常规分析和筛查应用。
