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Next-generation Sequencing

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....

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Related Experiment Video

Updated: May 24, 2025

VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma

VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma

Published on: December 28, 2015

Combined next generation sequencing for HLA typing and chimerism testing.

Jennifer Tyler¹, Heather Casey¹, Carrie Mowery¹

¹Department of Pathology, Penn State Hershey Medical Center, Hershey, PA, United States.

Human Immunology

|March 5, 2025

Summary

This summary is machine-generated.

Combining HLA typing and chimerism testing using next-generation sequencing (NGS) provides comparable results to separate assays. This integrated approach offers potential benefits for clinical diagnostics and research applications.

Keywords:

Flow cell NGS Read length Sample number

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VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma

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Personalized Peptide Arrays for Detection of HLA Alloantibodies in Organ Transplantation

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Detection of Residual Donor Erythroid Progenitor Cells after Hematopoietic Stem Cell Transplantation for Patients with Hemoglobinopathies

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Published on: September 6, 2017

Area of Science:

Molecular Biology
Genetics
Clinical Diagnostics

Background:

Next-generation sequencing (NGS) enables multiplexing of molecular assays.
HLA typing and chimerism testing are crucial for transplantation and diagnostics.

Purpose of the Study:

To evaluate the feasibility and performance of combining HLA typing and chimerism testing in a single NGS workflow.
To assess the benefits and considerations of integrated NGS assays.

Main Methods:

Two NGS-based assays (HLA typing and chimerism testing) were combined and sequenced using a MiSeq V2 reagent kit.
Library pools were combined at specific volume ratios, and sample sheets were optimized for dual indexing and 150 bp read length.
FASTQ files were generated and analyzed on the BaseSpace Sequence Hub.

Main Results:

Both combined and separate NGS approaches yielded sufficient read numbers and high sequence quality.
The integrated assay demonstrated a chimerism testing sensitivity of 0.1%.
Comparable test results were achieved for clinical samples in both HLA typing (N=44) and chimerism genotyping (N=24).

Conclusions:

Combining HLA typing and chimerism testing via NGS is a viable strategy.
The integrated approach maintains assay performance and offers potential efficiencies for clinical applications.
Further evaluation of benefits and considerations for combined NGS tests is warranted.