ArticleJournal of clinical microbiology1998
Comparative study of different standardization concepts in quantitative competitive reverse transcription-PCR assays.
Article in Journal of clinical microbiology, 1998. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed.
- Determination of the most suitable reference gene for transcription analysis of synovial tissue from osteoarthritis patients.Scientific reports · 2026Article
- Assessment of reference genes for qRT-PCR normalization to elucidate host response to African swine fever infection.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2024Article
- Nanoparticle-Shielded dsRNA Delivery for Enhancing RNAi Efficiency in Cotton Spotted BollwormInternational journal of molecular sciences · 2023Article
- Identification of stable reference genes for quantitative gene expression analysis in the duodenum of meat-type ducks.Frontiers in veterinary science · 2023Article
- Normalization of Gene Expression by Quantitative RT-PCR in Human Cell Line: comparison of 12 Endogenous Reference Genes.Ethiopian journal of health sciences · 2018Article
- Photodynamic inactivation in the expression of the Candida albicans genes ALS3, HWP1, BCR1, TEC1, CPH1, and EFG1 in biofilms.Lasers in medical science · 2018Article
- Validation of Reference Genes for Expression Studies in Human Meningiomas under Different Experimental Settings.Molecular neurobiology · 2018Article
- Selection of housekeeping genes and demonstration of RNAi in cotton leafhopper, Amrasca biguttula biguttula (Ishida).PloS one · 2018Article
- Article
- Identification of suitable endogenous controls for gene and miRNA expression studies in irradiated prostate cancer cells.Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine · 2015Article
- Evaluation of endogenous control gene(s) for gene expression studies in human blood exposed to 60Co γ-rays ex vivo.Journal of radiation research · 2015Article
- Application of qRT-PCR and RNA-Seq analysis for the identification of housekeeping genes useful for normalization of gene expression values during Striga hermonthica development.Molecular biology reports · 2013Article
- Application of quantitative PCR for the detection of microorganisms in water.Analytical and bioanalytical chemistry · 2013Review
- Identification of stable endogenous control genes for transcriptional profiling of photon, proton and carbon-ion irradiated cells.Radiation oncology (London, England) · 2012Article
- Reference gene selection for normalization of PCR analysis in chicken embryo fibroblast infected with H5N1 AIV.Virologica Sinica · 2010Article
- Reference gene selection for head and neck squamous cell carcinoma gene expression studies.BMC molecular biology · 2009Article
- Selection of housekeeping genes for gene expression studies in human reticulocytes using real-time PCR.BMC molecular biology · 2006Article
- Selection of reference genes for gene expression studies in human neutrophils by real-time PCR.BMC molecular biology · 2005Article
- Applications of competitor RNA in diagnostic reverse transcription-PCR.Journal of clinical microbiology · 2003Article
- Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR.Nucleic acids research · 2002Article
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Four different standardization approaches based on a competitive reverse transcription (RT)-PCR assay were compared with a noncompetitive assay based on an external standard curve. Criteria for assessment were accuracy in quantitation, correctness of recovery, sensitivity, dynamic range, reproducibility, throughput, and convenience of sample handling. As a model system, we used the 5'-noncoding region of hepatitis C virus (HCV) for amplification in all quantitative RT-PCRs. A computer program that allowed parallel data processing was developed. Surprisingly, all methods were found suitable for accurate quantitation and comparable with respect to the criterion correctness of recovery. All results differed only by a factor of about 2. The reason for this finding might be that all of our mimics, as well as the wild-type genome of HCV, exhibited exactly the same amplification and hybridization efficacy. Moreover, minimal competition occurred in our experiments over a 5-log dynamic range. A further topic of our investigation was the comparison of two different competitive RNA fragments, mimics, with regard to their suitability as internal standards. One was a heterologous mimic, in which only the primer binding sites were identical to the wild type. The second one was a homologous mimic identical to the wild type except for a small region used for differential hybridization, which was replaced by a permutated sequence of the same length. Both the homologous and heterologous internal mimics were found appropriate for an accurate competitive RT-PCR assay, provided that amplification efficacy, as well as capture efficacy, is proven identical for both analyte and mimic.
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