High-Resolution Full-Length HLA Typing Method Using Third Generation (Pac-Bio SMRT) Sequencing Technology
The human HLA genes are among the most polymorphic genes in the human genome. Therefore, it is very difficult to find two unrelated individuals with identical HLA molecules. As a result, HLA Class I and Class II genes are routinely sequenced or serotyped for organ transplantation, autoimmune disease-association studies, drug hypersensitivity research, and other applications. However, these methods were able to give two or four digit data, which was not sufficient enough to understand the completeness of haplotypes of HLA genes. To overcome these limitations, we here described end-to-end workflow for sequencing of HLA class I and class II genes using third generation sequencing, SMRT technology. This method produces fully-phased, unambiguous, allele-level information on the PacBio System.
KeywordsSingle molecule Real time Sequencing HLA typing PacBio High resolution Full length
We are thankful to Dr. Sudhir Krishna, NCBS for his scientific guidance and funding this research. We thank Dr. Latha Jagannathan and Dr. Nutan Dighe, Bangalore Medical Services Trust, Bangalore for their constant support and providing the samples for research work. We thank Mohammad Zahid from Shiva Scientific/GenDx for providing PacBio HLA primers. Thanks to Dr. Anil Singh and Mr. Mohit from Institute of Himalayan Bioresource Technology, Palampur and also thanks to Paras Yadav, Imperial Life Science (P) LTD., Delhi for their help to access PacBio sequencer. We appreciate Centre for Cellular and Molecular Platforms and Department of Biotechnology, Government of India for their support for this project.
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