Comparative analysis of NGS and sanger sequencing methods for HLA typing at a Russian university clinic

Abstract

The database of the World Health Organization (WHO) Nomenclature Committee for Factors of the HLA System (IPD-IMGT/HLA Database) contained information on the nucleotide sequences of 20272 different HLA alleles in September 2018, of which 14800 were HLA class I and 5288 were found for the HLA class II alleles. Over the last 20 years, the automated Sanger technique is a prevalent approach to genome sequencing in humans, animals, bacteria, and viruses. However, a need for more rapid routine genome screening stimulated novel technologies of multiplex DNA sequencing. These modern methods are depicted as the second-generation approaches (Next-Generation Sequencing, NGS). The aim of our research was a comparison of two methods and their efficiency evaluation. To achieve our purpose, we selected a group of 35 DNA samples, mainly from potential hematopoietic cells donors, and conducted a comparative analysis by Sanger and NGS method. NGS method allowed detecting rare or novel variants of alleles. This approach is confirmed to be more sensitive and more cost-effective, especially in large HLA-typing laboratories.