Abstract JBZ Den Bosch alpha-globin EMMD 2017

During EMMD 2017 Noordwijk, Jeroen Bosch Hospital will be present with their new abstract about α-thalassemia from Viennalab.

Good performance of the α-Globin StripAssay® for diagnostics of alphathalassemia

Ilse A.M. Luijten-de Vrije1 , Henk Martens2 , Jacqueline Leuvenink2 , Cornelis J.J (Ronald) Huijsmans2 , Adriaan J.C. van den Brule1 1 Jeroen Bosch Hospital, Laboratory for Molecular Diagnostics 2 Jeroen Bosch Hospital, Laboratory of Clinical Chemistry and Haematology

Alpha-thalassemia is a blood disorder characterized by a decrease in α-globin chain production due to deletion or mutation of one or more of the four α-globin genes. In this study we evaluated a commercial reverse-hybridisation strip assay in comparison to our in-house conventional PCR. A cohort of 40 archival DNA samples, extracted from EDTA-blood using either the QIAamp DNA mini kit (QIAGEN) or MagNA Pure 96 (Roche), was selected.

All samples were previously tested for the presence of 5 α-Globin deletions (-α 3.7 , -α 4.2 , – – MED , — SEA ,-(α)20.5) using a set of conventional in-house PCRs. The cohort was subsequently re-evaluated using the α-Globin StripAssay® (ViennaLab, Goffin Molecular Technologies) detecting 21 α-globin mutations: 7 deletions, the αααanti-3.7 gene triplication, two α1 and eleven α2 point mutations. Data interpretation was performed both manually and by using the StripAssay® evaluator software v2 (ViennaLab). Of the 40 samples, 16 were α-Globin wildtype in both assays. In another 16 samples, the 2 assays identified the same deletions. Five samples showed additional mutations using the StripAssay® , that were not covered in our in-house assay (– FIL , αααanti-3.7 gene triplication and α2 IVS1-5nt (n=3)). Finally, 3 samples were found invalid in the StripAssay® , most likely due to no adequate NA input . In one of these samples, results from the in-house assay indicated -α 3.7/-α 4.2, whilst the remaining 2 were wildtype. Re-analysis is presently being performed.

The StripAssay® showed good concordance with our in-house PCR assay and detected several additional α-Globin mutations because of its more comprehensive mutational coverage. StripAssay® is user friendly and less labour intensive than our in-house assay, especially in combination with the evaluator software.

 

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