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Journal of Translational Medicine
Published in: Journal of Translational Medicine 1/2003

Open Access 01-06-2003 | Methodology

Pyrosequencing™ : A one-step method for high resolution HLA typing

Authors: Daniel Ramon, Megan Braden, Sharon Adams, Francesco M Marincola, Lu Wang

Published in: Journal of Translational Medicine | Issue 1/2003

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Abstract

While the use of high-resolution molecular typing in routine matching of h uman l eukocyte a ntigens (HLA) is expected to improve unrelated donor selection and transplant outcome, the genetic complexity of HLA still makes the current methodology limited and laborious. Pyrosequencing™ is a gel-free, sequencing-by-synthesis method. In a Pyrosequencing reaction, nucleotide incorporation proceeds sequentially along each DNA template at a given n ucleotide d ispensation o rder (NDO) that is programmed into a pyrosequencer. Here we describe the design of a NDO that generates a pyrogram unique for any given allele or combination of alleles. We present examples of unique pyrograms generated from each of two heterozygous HLA templates, which would otherwise remain cis/trans ambiguous using standard s equencing b ased t yping (SBT) method. In addition, we display representative data that demonstrate long read and linear signal generation. These features are prerequisite of high-resolution typing and automated data analysis. In conclusion Pyrosequencing is a one-step method for high resolution DNA typing.
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Metadata
Title
Pyrosequencing™ : A one-step method for high resolution HLA typing
Authors
Daniel Ramon
Megan Braden
Sharon Adams
Francesco M Marincola
Lu Wang
Publication date
01-06-2003
Publisher
BioMed Central
Published in
Journal of Translational Medicine / Issue 1/2003
Electronic ISSN: 1479-5876
DOI
https://doi.org/10.1186/1479-5876-1-9

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