HLA typing

We provide a number of professional Laboratory Services at Anthony Nolan, including a gold-standard Human Leukocyte Antigen (HLA) Typing service in line with European standards.  HLA typing is used to match patients and donors for transplantation. It is also helpful for researchers working in areas of preclinical research, such as immunotherapy.

While understanding of the importance of HLA is growing, the actual process of HLA typing is complex and needs to be performed by highly competent and skilled specialists. Many hospitals and researchers in academia and industry therefore opt to outsource HLA typing to expert laboratories, such as Anthony Nolan.  

If you have a specific HLA typing request, complete our Contact Us form and we’ll aim to get back to you within one working day. Otherwise, read on to find out more about our service, processes and the options available to you.

What is HLA?

HLA molecules are made up of multiple proteins, and our genes carry the instructions to make each of them. Altogether, there are tens of thousands of different versions of these genes, known as alleles, resulting in the potential for many different combinations of HLA.  

HLA typing involves ‘reading’ the DNA sequence of HLA alleles.

Why is HLA typing helpful?

HLA typing is used to match potential donors with patients in need of a transplant with the aim of providing the best possible match - and therefore the best possible outcome.

HLA typing can also be used to help diagnose some conditions, and predict how someone may react to certain types of medicines, for example:

• Disease association: There are certain HLA alleles which are linked to immune conditions like coeliac disease and ankylosing spondylitis, so HLA typing can be used to help with their diagnosis.
• Pharmacogenetics: A person’s HLA alleles can influence how they respond to medications. For example, people who are HIV-positive are sometimes treated with a specific drug. But it’s long been known that this drug causes severe side-effects in those who carry an HLA allele called HLA-B*57:01. Testing for this particular allele therefore helps with clinical decision making.

Scientific understanding of the influence of HLA in medicine is growing all the time, and Anthony Nolan’s work is central to much of the pioneering research that’s happening in the sector.

Why Anthony Nolan?

Across the world, more than 35,000 different HLA alleles have already been identified, and new ones are being discovered all the time. The HLA research community is built on international collaboration between labs worldwide, so whenever a potential new HLA allele is discovered, the DNA sequence is submitted to one central database (the IPD-IMGT/HLA database). 

This critical database, containing sequences of all the HLA alleles that have ever been found, is maintained by Anthony Nolan’s Prof Steven Marsh and his Bioinformatics team.  So, if you need an HLA typing service, it makes sense to come to the industry leader: Contact Us now, for further information.

Our HLA typing services

HLA typing can be used for many applications, including but not limited to:

• Supporting patient/donor transplantation
• Anti-HLA antibody screening and identification
• Disease diagnosis, for conditions such as Coeliac, Ankylosing Spondylitis and Narcolepsy
• Pharmacogenetics and other hypersensitivity reactions
• Genetic mutation testing, e.g., HFE (Haemochromatosis risk) and CCR5 (HIV resistance) mutations

At Anthony Nolan, we perform genotyping of the following HLA genes: HLA-A, HLA-B, HLA-C, HLA-DRB1, HLA-DRB3, HLA-DRB4, HLA-DRB5, HLA-DQA1, HLA-DQB1, HLA-DPA1, HLA-DPB1.

We can also report HLA typing data in a variety of different formats to suit your needs, and are happy to help interpret your results if required. 

Not sure which level of HLA typing is best suited to your project, or need clarification around the complexities of HLA nomenclature? Contact Us and we’ll aim to get back to you within one working day. We can offer a completely bespoke service to help ensure you achieve the best outcomes, within the desired timeframe, and at a competitive price.

Our processes

Many of our clients choose to outsource their HLA typing requirements to Anthony Nolan, safe in the knowledge that we’re leaders in our field. If you’d like to know more about how HLA typing works, read on…

Overview: We analyse the DNA sequence of each HLA gene, using a state-of-the-art method known as Next Generation Sequencing (NGS), to accurately define specific alleles and determine a person’s HLA type.  The whole process takes 3-4 days from receiving the DNA samples to generating the results, and here’s why…

• Our step-by-step process:

First, DNA samples – the patient or donor samples, plus control samples – are extracted and prepared by our Technical Services Team to confirm they are suitable for downstream processes. We then use Next generation sequencing (NGS) to determine the DNA sequence of the HLA genes from a sample such as blood, saliva or swab.

The process is illustrated above and takes place over 3-4 days:

• Day 1: 

Amplification and Visualisation 

We start by amplifying 11 HLA genes. These are HLA-A, -B, -C, -DRB1, -DRB3, -DRB4, -DRB5, -DQA1, -DQB1, -DPA1 and -DPB1. The samples are amplified using a process called PCR (polymerase chain reaction), which creates millions of copies of the desired HLA genes for each sample. We use fluorescent dyes to visualise successful DNA amplification.

• Day 2: 

Library preparation, from DNA pooling to sequencer loading

During PCR, the genes of interest within each sample are amplified separately. The next stage is to pool - or combine - the genes. The proportions of the pooled DNA are determined by quantifying each amplified gene. This is the beginning of a stage that is known as ‘library preparation’. It results in all our samples being uniquely tagged in one tube, ready to be loaded onto an Illumina flow cell (a hollow glass slide used to place tagged libraries into the sequencer).

• Day 3 / 4: 

Sequencing  

The flow cell is critical in sequencing. It is the conduit on which DNA strands are replicated to form lots of DNA templates for sequencing by synthesis. This process utilizes fluorescent bases made of dNTPs (artificial nucleotides). As each individual DNA base (A, C, T or G – the four different chemicals that make up all our DNA) is added to the new DNA strand, it is detected by the MiSeq sequencing instrument. The DNA sequence for each sample is then recorded in files known as FASTQ.

Data interpretation

The FASTQ files are processed and sorted by a program called NGSengine. A scientist can then analyse and review the data, accurately identifying 11 genes of high to allelic resolution HLA typing for our samples. This level of accuracy is helpful in selecting the best-matched donor for transplants, and in preclinical research, particularly that examining immune responses to disease.

Working in partnership

Anthony Nolan holds the world’s only central database, charting the 35,000+ different HLA alleles that have been identified to date. This attests to our leading position within the Histocompatibility and Immunogenetics community, and makes us the ideal partner for researchers and developers requiring HLA typing services.

Whatever your requirements, we can provide a flexible service tailored to your exact brief, all delivered with the level of expertise and service you’d expect from HLA specialists. You also have the reassurance that the Anthony Nolan Histocompatibility Laboratories are accredited by United Kingdom Accreditation Service (UKAS) to ISO15189 and the European Federation for Immunogenetics (EFI).

Contact us to find out more.