Document Detail


Single nucleotide polymorphism-based system improves the applicability of quantitative PCR for chimerism monitoring.
MedLine Citation:
PMID:  19056844     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Egle Gineikiene; Mindaugas Stoskus; Laimonas Griskevicius
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Publication Detail:
Type:  Journal Article     Date:  2008-12-04
Journal Detail:
Title:  The Journal of molecular diagnostics : JMD     Volume:  11     ISSN:  1525-1578     ISO Abbreviation:  J Mol Diagn     Publication Date:  2009 Jan 
Date Detail:
Created Date:  2008-12-30     Completed Date:  2009-05-05     Revised Date:  2010-09-23    
Medline Journal Info:
Nlm Unique ID:  100893612     Medline TA:  J Mol Diagn     Country:  United States    
Other Details:
Languages:  eng     Pagination:  66-74     Citation Subset:  IM    
Affiliation:
Hematology, Oncology, and Transfusion Medicine Center, Vilnius University Hospital Santariskiu Clinics, Vilnius, Lithuania. egle.gineikiene@santa.lt
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MeSH Terms
Descriptor/Qualifier:
Alleles
Bone Marrow Transplantation
Chimerism*
DNA Primers / genetics
Genetic Markers*
Genotype
Humans
Polymerase Chain Reaction / methods*
Polymorphism, Single Nucleotide*
Chemical
Reg. No./Substance:
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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