HLA(NGS) - CB-Gene

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HLA (NGS) Testing Services

Human leukocyte antigens (HLA) encode genes for the major histocompatibility complex (MHC) in the human body and play a key role in immune responses. The high polymorphism of HLA genes makes them of great significance in organ transplantation, autoimmune disease research, and pharmacogenomics. Traditional HLA typing methods suffer from limitations such as low resolution and limited throughput. The emergence of next-generation sequencing (NGS) technology has revolutionized HLA typing.

Definition

NGS-HLA typing uses high-throughput sequencing technology to sequence the entire HLA gene, achieving high-resolution and high-accuracy typing. Compared to traditional methods (such as PCR-SSO, PCR-SSP, and Sanger sequencing), NGS can cover the entire HLA gene region, including exons, introns, and untranslated regions, providing more comprehensive genetic information.

Principle

NGS technology uses massively parallel sequencing to simultaneously sequence millions of DNA fragments. In HLA typing, the target HLA gene regions are first enriched using PCR or hybridization capture technology. Sequencing libraries are then constructed and sequenced on a high-throughput sequencing platform. The resulting massive data sets undergo bioinformatics analysis and are compared with a database of known HLA alleles to accurately determine the HLA type of the sample.

Workflow

Sample Collection and DNA Extraction

High-quality genomic DNA is extracted from blood or saliva.

Target Region Enrichment

Multiplex PCR or probe capture technology is used to specifically amplify the HLA gene regions.

Library Construction

Sequencing adapters are added to the amplified products to construct a sequencing-ready library.

High-Throughput Sequencing

Sequencing is performed on platforms such as Illumina and Ion Torrent, generating short or long reads.

Data Analysis

Sequence alignment, allele assignment, and result interpretation are performed using specialized software (such as TypeStream and HLA-VBSeq).

Report Generation

Detailed typing results, including allele nomenclature and quality control information, are provided.

Advantages

High Resolution

Able to distinguish subtle variants that are undetectable by traditional methods, providing four-digit or even eight-digit allele designations. .

High Throughput

Multiple samples can be tested in a single run, significantly improving efficiency and reducing costs.

Comprehensiveness

Covers the entire HLA gene region, including highly polymorphic regions that were previously difficult to analyze.

Accuracy

Parallel sequencing and redundant coverage reduce false positives and enhance the reliability of results.

Automation

Standardized processes from sample processing to data analysis mitigate human error.

Application Scenarios

Organ and Hematopoietic Stem Cell Transplantation: High-resolution HLA typing is the gold standard for transplant matching, reducing the risk of graft-versus-host disease (GVHD) and rejection.

Autoimmune Disease Research: HLA genes are closely associated with diseases such as rheumatoid arthritis and ankylosing spondylitis. NGS can help discover new susceptibility alleles.

Pharmacogenomics: Certain HLA alleles are associated with adverse drug reactions (such as Stevens-Johnson syndrome caused by carbamazepine). NGS typing can guide personalized medication.

Infectious Disease Research: HLA polymorphisms influence immune responses to viruses such as HIV and hepatitis B. Studying HLA can aid in vaccine design and the exploration of disease mechanisms.

Forensic Medicine and Human Genetics: HLA typing can be used for paternity testing and the study of genetic diversity in populations.

The introduction of NGS technology has ushered in a new era for HLA typing, providing powerful tools for clinical transplantation, disease research, and personalized medicine. As technology continues to mature and costs decrease, NGS-HLA typing will become an important part of routine diagnosis and promote the development of precision medicine.

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