Instruments & Reagents

Introduction

Human leukocyte antigen (HLA) genotyping is a critical procedure for donor selection in organ and hematopoietic stem cell transplantation. The resolution of HLA genotyping directly dictates the level of histocompatibility between donors and recipients, thereby influencing the risk of transplant rejection and overall success rates.

The AltruType^TM Long-Read Sequencing HLA Genotyping Kit (Single-Molecule Sequencing Method) leverages advanced long-read sequencing technology to achieve high-resolution genotyping of multiple HLA genes. It provides robust, professional support for clinical donor screening, as well as diagnostic and therapeutic applications in transplantation.

Intended Purpose

This kit is intended for the identification and characterization of HLA complex genes from human genomic DNA isolated from whole blood. It targets Class I (classical HLA-A, HLA-B, HLA-C) and Class II (HLA-DQA1, HLA-DQB1, HLA-DRB1, HLA-DRB3/4/5, HLA-DPA1, HLA-DPB1) genes.

The workflow utilizes polymerase chain reaction (PCR) to amplify a predefined set of target genes. The resulting amplicons are designed for downstream library preparation and sequencing on PacBio HiFi platforms. The generated sequencing data is subsequently analyzed using Haorui TransMatch^TM software to produce high-resolution HLA genotyping results.

The assay provides an HLA profile of the tested individual, intended to assist in:

(1) Assessing HLA gene compatibility between patients and potential donors for transplantation.

(2) Diagnosing autoimmune diseases.

(3) Predicting adverse drug reactions.

Note: Test results obtained with this kit are for clinical reference only and should not be used as the sole basis for diagnosis or clinical decision-making. Clinicians should make comprehensive judgments and treatment decisions by integrating these results with the patient’s medical history, physical examination, and other laboratory findings.

Principle of the Method

The AltruType^TM Long-Read Sequencing HLA Genotyping Kit (available in 24/48/96/192 reactions) is an integrated solution designed for the amplification and library preparation of HLA targets. The kit enables the simultaneous amplification of 11 HLA loci (HLA-A, -B, -C, -DQA1, -DQB1, -DPA1, -DPB1, -DRB1, and -DRB3/4/5) followed by streamlined library construction for sequencing.

The workflow proceeds as follows:

(1) Nucleic Acid Extraction: Genomic DNA is isolated from the sample.

(2) PCR Enrichment: Target regions are amplified via a single, long-range multiplex PCR covering all 11 loci.

(3) Library Preparation: PacBio’s proprietary SMRTbell adapters are ligated to the amplicons to construct circular DNA libraries.

(4) Sequencing: Libraries are loaded onto the Revio or Vega long-read sequencing platform using the HiFi sequencing mode.

(5) Signal Detection: Fluorescently labeled nucleotides are incorporated during polymerization, and real-time signals are captured to generate high-fidelity (HiFi) reads.

(6) Data Analysis: Reads are aligned to the human genome and HLA reference sequences (IPD-IMGT/HLA Database) using Haorui TransMatch software to export high-resolution HLA genotyping results.

Product Configuration

Shipping and Storage

1. Storage Conditions

Store the kit at -20 ±5°C. The shelf life is 3 months from the date of manufacture (Note: Magnetic beads should be stored in a 4°C refrigerator).

2. Freeze-Thaw Stability

The kit may undergo no more than 6 freeze-thaw cycles. Once opened, use immediately.

3.  Shipping Requirements

Transport under temperature-controlled conditions (-20 ±5°C).

Maximum allowable transit time: 3 days.

Upon receipt, verify that the internal temperature remains within -20 ±5°C before opening.

4. Refer to the lot-specific label for manufacturing date (MFG) and expiration date (EXP).

For general inquiries, orders, pricing, delivery, or product usage information, please reach out to  us via email: marketing@xahorizon.cn.