PB1381 - Value of next-generation sequencing in the diagnosis of platelet function disorders

Tuesday, June 27, 2023

18:30 – 19:30 ET

Room: Exhibition

Co-Author(s)

RG

Réka Gindele

assistant lecturer
Division of Clinical Laboratory Science, Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
Debrecen, Hajdu-Bihar, Hungary
AK

Adrienne Kerényi

senior lecturer
Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
Debrecen, Hajdu-Bihar, Hungary
ZS

Zsuzsanna Szűcs

assistant research fellow
Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
Debrecen, Hajdu-Bihar, Hungary
LM

László Madar

assistant research fellow
Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
Debrecen, Hajdu-Bihar, Hungary
ZS

Zsuzsanna Szabó

technician
Division of Clinical Laboratory Science, Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
Debrecen, Hajdu-Bihar, Hungary
AI

Angelika Illés

trainee
Division of Clinical Laboratory Science, Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
Debrecen, Hajdu-Bihar, Hungary
IB

István Balogh

professor, head of division
Division of Clinical Genetics, Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
Debrecen, Hajdu-Bihar, Hungary

Presenting Author(s)

Zsuzsanna Bereczky, MD, PhD (she/her/hers)

head of division
Division of Clinical Laboratory Science, Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
Debrecen, Hajdu-Bihar, Hungary

Background: Diagnosis of platelet disorders is sometimes challenging and there are also several differential diagnostics issues. Next-generation sequencing (NGS) is a useful tool to overcome these problems. However, in this group of diseases, there are still few literature data available obtained by this methodology.

Aims: To analyse the value of NGS testing in platelet disorders in cases with well-characterised clinical and laboratory phenotype in a tertiary clinical center.

Methods: We selected 45 patients with probable hereditary platelet disorders or von Willebrand disease based on their clinical and laboratory results. DNA samples were isolated from peripheral blood samples, and then NGS analysis was performed. A custom designed panel was used including 14 genes (ADRA2A, FLI1, GFI1B, GNAI1, GNAQ, NBEA, NBEAL2, P2RY1, P2RY12, PLA2G4A, PLAU, PLCB2, RASGRP2 , STXBP2) for platelet disorders and another one was applied in cases with suspected von Willebrand disease. In whom no mutations were found by panel sequencing (n=15) clinical exome sequencing was performed. Using this platform we were able to analyse 4,727 genes. Sanger sequencing was performed with SeqStudio Genetic Analyzer to verify the detected mutations. In order to assess the pathogenicity of new genetic variations we used MutPred2, PolyPhen-2, SIFT, MutationTaster, VarSome and Franklin applications.

Results: By using the custom panel, we were able to detect genetic variations in a total of 30 patients (66%). In addition, the occurrence of synonymous mutations was very common. Out of the 15 patients who underwent a successful clinical exome analysis, we detected genetic variations in 10 cases (66.7%), which correlated with the patients' laboratory and clinical phenotype.

Conclusion(s): The NGS genetic analysis seems to be a valuable tool in the diagnosis of platelet function disorders and genotype-phenotype correlations can be established. Association of novel variants with clinical and laboratory characteristics provides useful pieces of information in the diagnostic process.