Application of High-Throughput Sequencing in the Diagnosis of Inherited Thrombocytopenia

Abstract

Inherited thrombocytopenia is a group of hereditary diseases with a reduction in platelet count as the main clinical manifestation. Clinically, there is an urgent need for a convenient and rapid diagnosis method. We introduced a high-throughput, next-generation sequencing (NGS) platform into the routine diagnosis of patients with unexplained thrombocytopenia and analyzed the gene sequencing results to evaluate the value of NGS technology in the screening and diagnosis of inherited thrombocytopenia. From a cohort of 112 patients with thrombocytopenia, we screened 43 patients with hereditary features. For the blood samples of these 43 patients, a gene sequencing platform for hemorrhagic and thrombotic diseases comprising 89 genes was used to perform gene detection using NGS technology. When we combined the screening results with clinical features and other findings, 15 (34.9%) of 43patients were diagnosed with inherited thrombocytopenia. In addition, 19 pathogenic variants, including 8 previously unreported variants, were identified in these patients. Through the use of this detection platform, we expect to establish a more effective diagnostic approach to such disorders.

Strengths and limitations of high‐throughput sequencing for the diagnosis of inherited bleeding and platelet disorders

“…Different HTS studies for diagnostics of BPD have been published but for this review, we have only focussed on the studies that have used the guidelines for variant classification as formulated by the American College of Medical Genetics (ACMG) in 2015 [10]. Figure 2 provides an overview of the diagnostic rates obtained in these studies that however differ in terms of used HTS methods (TS or WES), studied pathologies (platelet disorders, coagulation disorders and bleeding of unknown etiology) and the number of genes and patients included [11][12][13][14][15][16][17][18][19][20][21]. In addition, these studies have used different inclusion criteria ranging from the selection of only patients with a known or suspected etiology (studies with typically high diagnostic rates) to patients with bleeding of unknown etiology and normal laboratory parameters (studies with very low diagnostic rates).…”

“…Other HTS studies have also reported variants in more than one gene but without further specifications if these had a clinical impact for the patient or family [11,[14][15][16][17][18]20]. Such examples of mostly digenic inheritance have recently also been documented in case reports.…”

“…10 Figure 2 provides an overview of the diagnostic rates obtained in these studies that however differ in terms of used HTS methods (TS or WES), studied pathologies (platelet disorders, coagulation disorders, and bleeding of unknown etiology), and the number of genes and patients included. [11][12][13][14][15][16][17][18][19][20][21] Most of these HTS studies provided no detailed information related to the detection of CNV. [11][12][13][14][15][16][17][18][19][20][21] The ThromboGenomics study reported a CNV in 40 of the 2396 tested patients; these were predicted to affect single exons (n = 11), multiple exons (n = 15), or whole genes (n = 14).…”

Diagnosis of Inherited Platelet Disorders on a Blood Smear

“…The spread of next-generation sequencing (NGS) greatly contributed to the unraveling of the genetic basis of several forms of IPD, and thus immediately became a significant diagnostic tool in the field. Using a small amount of blood with minor risk of pre-analytical artifacts, it can provide patients with a highly accurate diagnosis [4][5][6][7][66][67][68][69][70][71][72]. As it becomes cheaper to perform, it is not inconceivable that NGS could become the standard above other diagnostic techniques.…”

Inherited Thrombocytopenias: Combining High-Throughput Sequencing With Other Relevant Data

“…We read with interest the article by Wang et al recently published in Clinical and Applied Thrombosis/Hemostasis . 1 Here, the authors have evaluated the role of high-throughput sequencing in 43 apparently familial cases of thrombocytopenia using Drachman 2 review algorithm.…”