| Single nucleotide polymorphism-based system improves the applicability of quantitative PCR for chimerism monitoring. | |
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MedLine Citation:
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PMID: 19056844 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Recently, several studies demonstrated the feasibility of a real-time quantitative PCR (qPCR) approach for chimerism monitoring. qPCR offers a fast, sensitive, and elegant quantification of genotypes. However, before it becomes an established method for routine chimerism monitoring, a qPCR marker set for every transplant pair should be available. This requirement poses a major challenge since the genetic markers for qPCR--short insertions/deletions (Indels) and single nucleotide polymorphisms (SNPs)--published to-date do not guarantee applicability for every transplant pair. The aim of our study was to design and validate a new SNP allele-specific system to supplement an already existing Indel primer panel and improve applicability of the qPCR approach for chimerism status monitoring. Here, we present an approach for an economical in-house design of SNP allele-specific qPCR primers/probe sets with a locus-individualized reference system that allows for the accurate quantification of the respective informative locus using a simple DeltaDeltaCt method. We designed primers/probe sets specific for seven biallelic SNP loci and validated them in a population of 30 transplant pairs. Repeatability varied depending on the amount of quantifiable genotype. The combination of our SNP-qPCR system and Indel primers increased recipient genotype identification from 86.6% to 96.6% when tested in a population of our transplant pairs. These results demonstrate the feasibility of our SNP-based qPCR approach to improve the applicability of a qPCR for chimerism monitoring. |
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Authors:
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Egle Gineikiene; Mindaugas Stoskus; Laimonas Griskevicius |
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Publication Detail:
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Type: Journal Article Date: 2008-12-04 |
Journal Detail:
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Title: The Journal of molecular diagnostics : JMD Volume: 11 ISSN: 1525-1578 ISO Abbreviation: J Mol Diagn Publication Date: 2009 Jan |
Date Detail:
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Created Date: 2008-12-30 Completed Date: 2009-05-05 Revised Date: 2010-09-23 |
Medline Journal Info:
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Nlm Unique ID: 100893612 Medline TA: J Mol Diagn Country: United States |
Other Details:
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Languages: eng Pagination: 66-74 Citation Subset: IM |
Affiliation:
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Hematology, Oncology, and Transfusion Medicine Center, Vilnius University Hospital Santariskiu Clinics, Vilnius, Lithuania. egle.gineikiene@santa.lt |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Alleles Bone Marrow Transplantation Chimerism* DNA Primers / genetics Genetic Markers* Genotype Humans Polymerase Chain Reaction / methods* Polymorphism, Single Nucleotide* |
| Chemical | |
Reg. No./Substance:
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0/DNA Primers; 0/Genetic Markers |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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