ArticlePloS one2011
Accuracy in copy number calling by qPCR and PRT: a matter of DNA.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed.
- Effects of salivary amylase gene copy number on metabolic health.Frontiers in nutrition · 2026Review
- Review
- Frequent expansion of Plasmodium vivax Duffy Binding Protein in Ethiopia and its epidemiological significance.PLoS neglected tropical diseases · 2019Article
- A Review of Copy Number Variants in Inherited Neuropathies.Current genomics · 2018Review
- Quantifying mitochondrial DNA copy number using robust regression to interpret real time PCR results.BMC research notes · 2017Article
- Article
- Application of Droplet Digital PCR for Estimating Vector Copy Number States in Stem Cell Gene Therapy.Human gene therapy methods · 2016Article
- Article
- Identification of genomic aberrations in hemangioblastoma by droplet digital PCR and SNP microarray highlights novel candidate genes and pathways for pathogenesis.BMC genomics · 2016Article
- Association of Higher Defensin β-4 Genomic Copy Numbers with Behçet's Disease in Iraqi Patients.Sultan Qaboos University medical journal · 2015Article
- Quantitative high resolution melting: two methods to determine SNP allele frequencies from pooled samples.BMC genetics · 2015Article
- A pyrosequencing-based assay for the rapid detection of the 22q11.2 deletion in DNA from buccal and dried blood spot samples.The Journal of molecular diagnostics : JMD · 2014Article
- Technical considerations for genotyping multi-allelic copy number variation (CNV), in regions of segmental duplication.BMC genomics · 2014Article
- Comparison of pre-analytical FFPE sample preparation methods and their impact on massively parallel sequencing in routine diagnostics.PloS one · 2014Article
- Current analysis platforms and methods for detecting copy number variation.Physiological genomics · 2013Review
- Article
- Article
- De novo and inherited CNVs in MZ twin pairs selected for discordance and concordance on Attention Problems.European journal of human genetics : EJHG · 2012Article
Corrections and comments
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Authors and funding
8 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The possible implication of copy number variation (CNV) in the genetic susceptibility to human disease needs to be assessed using robust methods that can be applied at a population scale. In this report, we analyze the performance of the two major techniques, quantitative PCR (qPCR) and paralog ratio test (PRT), and investigate the influence of input DNA amount and template integrity on the reliability of both methods. Analysis of three genes (PRELID1, SYNPO and DEFB4) in a large sample set showed that both methods are prone to false copy number assignments if sufficient attention is not paid to DNA concentration and quality. Accurate normalization of samples is essential for reproducible qPCR because it avoids the effect of differential amplification efficiencies between target and control assays, whereas PRT is generally more sensitive to template degradation due to the fact that longer amplicons are usually needed to optimize sensitivity and specificity of paralog sequence PCR. The use of normalized, high quality genomic DNA yields comparable results with both methods.
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