ArticleScientific reports2017
Evaluation of TaqMan qPCR System Integrating Two Identically Labelled Hydrolysis Probes in Single Assay.
Article in Scientific reports, 2017. 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.
- Development of an Aptamer-Based qPCR Method for the Selective and Rapid Picomolar-Level Detection of Perfluorooctanesulfonic Acid in Water.Environmental science & technology · 2025Article
- Molecular advances in transfusion medicine: a narrative review.Frontiers in medicine · 2025Review
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- Potential and application of abortive transcripts as a novel molecular marker of cancers.World journal of experimental medicine · 2024Review
- Development of a highly sensitive TaqMan method based on multi-probe strategy: its application in ASFV detection.Biology methods & protocols · 2024Article
- Uncovering Molecular Quencher Effects on FRET Phenomena in Microsphere-Immobilized Probe Systems.Analytical chemistry · 2023Article
- A Multiplex PCR Melting-Curve-Analysis-Based Detection Method for the Discrimination of FiveJournal of fungi (Basel, Switzerland) · 2023Article
- A Novel, Precise and High-Throughput Technology for Viroid Detection in Cannabis (MFDetectViruses · 2023Article
- Review
- Divergent Synthesis of Ultrabright and Dendritic Xanthenes for Enhanced Click-Chemistry-Based Bioimaging.Chemistry (Weinheim an der Bergstrasse, Germany) · 2023Article
- Real-Time PCR Protocol for Detection and Quantification of Three Pathogenic Members of the Vibrionaceae Family.Microorganisms · 2022Article
- Sequential development of several RT-qPCR tests using LNA nucleotides and dual probe technology to differentiate SARS-CoV-2 from influenza A and B.Microbial biotechnology · 2022Article
- Systematic evaluation of TaqMan real-time polymerase chain reaction assays targeting theVeterinary world · 2022Article
- Development and Validation of Two Diagnostic Real-Time PCR (TaqMan) Assays for the Detection ofMicroorganisms · 2021Article
- Surveillance of SARS-CoV-2 lineage B.1.1.7 in Slovakia using a novel, multiplexed RT-qPCR assay.Scientific reports · 2021Article
- Developing, Modifying, and Validating a TaqMan Real-Time PCR Technique for Accurate Identification ofFrontiers in cellular and infection microbiology · 2021Article
- In Situ Profiling of the Three Dominant Phyla Within the Human Gut Using TaqMan PCR for Pre-Hospital Diagnosis of Gut Dysbiosis.International journal of molecular sciences · 2020Article
- Evaluation of EZplex MTBC/NTM Real-Time PCR kit: diagnostic accuracy and efficacy in vaccination.Clinical and experimental vaccine research · 2018Article
Corrections and comments
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Authors and funding
8 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Ongoing evolution of viral pathogens is a significant issue in diagnostic virology employing TaqMan qPCR/RT-qPCR. Specific concerns are related to false negativity due to probe binding failure. One option for compensating for such deficiency is to integrate a second identically labelled probe in the assay. However, how this alteration influences the reaction parameters has not been comprehensively demonstrated. In the present study, we evaluate a TaqMan protocol using two identically labelled hydrolysis probes (simple, LNA (locked-nucleic-acid)) and MGB (minor-groove-binder) modified probes and combinations thereof in a single assay. Our results based on a synthetic amplicon suggest that the second probe does not compromise the TaqMan qPCR/RT-qPCR parameters, which repeatedly and reproducibly remained comparable to those of the corresponding single-probe assays, irrespective of the relative probe orientation, whether opposite or tandem, and probe modifications or combinations thereof. On the other hand, the second probe additively contributed to the overall fluorescence signal. The utility of the dual-probe approach was demonstrated on practical examples by using field specimens. We hope that the present study might serve as a theoretical basis for the development or improvement of TaqMan qPCR/RT-qPCR assays for the detection of highly variable nucleic acid templates.
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