ArticleNucleic acids research2004
Sigmoidal curve-fitting redefines quantitative real-time PCR with the prospective of developing automated high-throughput applications.
Article in Nucleic acids research, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 81 papers.
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81 citing papers in PubMed.
- Implementation and Validation of a Limiting Component Quantification Method for qPCR.International journal of molecular sciences · 2026Article
- Quantification Revisited: What qPCR Efficiency Models Reveal About Data Analysis Integrity.International journal of molecular sciences · 2026Review
- A standard-curve-free ΔΔCt method for rapid GMO compliance screening relative to labeling thresholds.Frontiers in plant science · 2026Article
- Fabrication of Cost-Effective Microchip-Based Device Using Sandblasting Technique for Real-Time Multiplex PCR Detection.Micromachines · 2024Article
- Introducing the fBMC bioinformatics · 2024Article
- The TWW Growth Model and Its Application in the Analysis of Quantitative Polymerase Chain Reaction.Bioinformatics and biology insights · 2024Article
- Data treatment methods for real-time colorimetric loop-mediated isothermal amplification reactions.Scientific reports · 2023Article
- Next-generation molecular diagnostics: Leveraging digital technologies to enhance multiplexing in real-time PCR.Trends in analytical chemistry : TRAC · 2023Article
- PCR standard curve quantification in an extensive wastewater surveillance program: results from the Dutch SARS-CoV-2 wastewater surveillance.Frontiers in public health · 2023Article
- Adaptive Filtering Framework to Remove Nonspecific and Low-Efficiency Reactions in Multiplex Digital PCR Based on Sigmoidal Trends.Analytical chemistry · 2022Article
- A Point-of-Care Device for Fully Automated, Fast and Sensitive Protein Quantification via qPCR.Biosensors · 2022Article
- Evaluation validation of a qPCR curve analysis method and conventional approaches.BMC genomics · 2021Article
- Determination of Advantages and Limitations of qPCR Duplexing in a Single Fluorescent Channel.ACS omega · 2021Article
- Estimating Real-Time qPCR Amplification Efficiency from Single-Reaction Data.Life (Basel, Switzerland) · 2021Article
- Multiple imputation and direct estimation for qPCR data with non-detects.BMC bioinformatics · 2020Article
- Affine analysis for quantitative PCR measurements.Analytical and bioanalytical chemistry · 2020Article
- Critique of the pairwise method for estimating qPCR amplification efficiency: beware of correlated data!BMC bioinformatics · 2020Article
- Cryptosporidium parvum Elongation Factor 1α Participates in the Formation of Base Structure at the Infection Site During Invasion.The Journal of infectious diseases · 2020Article
- Framework for DNA Quantification and Outlier Detection Using Multidimensional Standard Curves.Analytical chemistry · 2019Article
- Simultaneous Single-Channel Multiplexing and Quantification of Carbapenem-Resistant Genes Using Multidimensional Standard Curves.Analytical chemistry · 2019Article
21 more citing papers are in PubMed but not listed here.
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Abstract
Quantitative real-time PCR has revolutionized many aspects of genetic research, biomedical diagnostics and pathogen detection. Nevertheless, the full potential of this technology has yet to be realized, primarily due to the limitations of the threshold-based methodologies that are currently used for quantitative analysis. Prone to errors caused by variations in reaction preparation and amplification conditions, these approaches necessitate construction of standard curves for each target sequence, significantly limiting the development of high-throughput applications that demand substantive levels of reliability and automation. In this study, an alternative approach based upon fitting of fluorescence data to a four-parametric sigmoid function is shown to dramatically increase both the utility and reliability of quantitative real-time PCR. By mathematically modeling individual amplification reactions, quantification can be achieved without the use of standard curves and without prior knowledge of amplification efficiency. Combined with provision of quantitative scale via optical calibration, sigmoidal curve-fitting could confer the capability for fully automated quantification of nucleic acids with unparalleled accuracy and reliability.
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