ArticleNucleic acids research2024
Advancing quantitative PCR with color cycle multiplex amplification.
Article in Nucleic acids research, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed.
- Dual-tetrahedral DNA-enabled strand displacement amplification biosensor for cell imaging and clinical detection of miR-34a-5p in NAFLD.Analytical and bioanalytical chemistry · 2026Article
- Analytical Validation and Preliminary Diagnostic Performance Evaluation of GenoPATHX™ Multiplex qPCR for Quantitative Detection of KeyPathogens (Basel, Switzerland) · 2026Article
- Programmable one-pot polymerase-mediated DNA synthesis via temperature control.Nature communications · 2026Article
- A novel duplex TaqMan qPCR assay for rapid differential diagnosis and co-infection screening of Orf virus and Brucella spp. in goats.Veterinary research communications · 2026Article
- Epidemiological surveillance of BVDV, Pasteurella multocida, and Brucella spp. infections in cattle from Guizhou, China using a newly established triplex TaqMan qPCR assay.Veterinary research communications · 2026Article
- Recognition, Signal Transduction, and Amplification of Nucleic Acids Under Isothermal Condition.Advanced genetics (Hoboken, N.J.) · 2026Article
- High-sensitivity, protein-independent detection of dsDNA sequences.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- Compartmentalization and Aggregation of Biomolecular Condensates in Crowded Hydrogels for Enhanced Nucleic Acid Diagnosis.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Development and epidemiological investigation of a TaqMan-based multiplex real-time quantitative PCR assay for simultaneous detection of five bovine viruses (BVDV, AKAV, BNoV, BEV, and BCoV).Frontiers in veterinary science · 2026Article
- Advances in methods for rapid detection of Staphylococcus aureus and methicillin-resistant S. aureus.Analytical sciences : the international journal of the Japan Society for Analytical Chemistry · 2025Review
- When Precision Matters: Bone Marrow Cytology Meets qPCR in a Pilot Study QuantifyingMicroorganisms · 2025Article
Corrections and comments
- Erratum issued
Authors and funding
11 authors.
Funding
Abstract
Quantitative PCR (qPCR) is the gold standard for detection and quantitation of known DNA targets, but the scarcity of spectrally distinct fluorophores and filter sets limits the number of detectable targets. Here, we introduce color cycle multiplex amplification (CCMA) to significantly increase the number of detectable DNA targets in a single qPCR reaction using standard instrumentation. In CCMA, presence of one DNA target species results in a pre-programmed pattern of fluorescence increases. This pattern is distinguished by cycle thresholds (Cts) through rationally designed delays in amplification. For example, we design an assay wherein Staphylococcus aureus sequentially induces FAM, then Cy5.5, then ROX fluorescence increases with more than 3 cycles between each signal. CCMA offers notably higher potential for multiplexing because it uses fluorescence permutation rather than combination. With 4 distinct fluorescence colors, CCMA theoretically allows the detection of up to 136 distinct DNA target sequences using fluorescence permutation. Experimentally, we demonstrated a single-tube qPCR assay screening 21 sepsis-related bacterial DNA targets in samples of blood, sputum, pleural effusion and bronchoalveolar lavage fluid, with 89% clinical sensitivity and 100% clinical specificity, showing its potential as a powerful tool for advanced quantitative screening in molecular diagnostics.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.