ArticleToxins2016
Comparison of Droplet Digital PCR and qPCR for the Quantification of Shiga Toxin-Producing Escherichia coli in Bovine Feces.
Article in Toxins, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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Who cites it
40 citing papers in PubMed.
- Challenges and Advances in the Detection of Leukemic Blasts in Cerebrospinal Fluid in Pediatric Acute Lymphoblastic Leukemia.Cancers · 2026Review
- Sensitivity of Loop-Mediated Isothermal Amplification in Comparison to Digital Droplet PCR for Identification ofFoods (Basel, Switzerland) · 2026Article
- Detection and colocalization of STEC genetic markers in wheat flour using whole cell digital droplet PCR.Frontiers in microbiology · 2026Article
- Development of a multiplex ddPCR assay for simultaneous absolute quantification of bacterial, fungal, and human DNA.PloS one · 2026Article
- Digital Droplet PCR (ddPCR) for Absolute Quantification of 16S rRNA Copy Number in Metagenomic Data.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Droplet digital PCR (ddPCR) for enhanced detection and quantification of bovine viral diarrhea virus (BVDV): A promising tool for disease control.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2025Article
- Detection of Norovirus from Berries in Serbia by Digital PCR and NGS.Foods (Basel, Switzerland) · 2025Article
- Development of loop-mediated isothermal amplification (LAMP) assays for the detection of diarrheagenicApplied and environmental microbiology · 2025Article
- Extirpation of Lake Sturgeon in an Ontario Lake Following Dam Construction and Watershed Diversion Confirmed by Indigenous Traditional Knowledge and Sedimentary eDNA.Ecology and evolution · 2025Article
- Optimizing C-Type Natriuretic Peptide and Receptor Expression Analysis with Droplet Digital™ PCR: Advancing Biomarker Discovery for Brugada Syndrome?Biomolecules · 2025Article
- Digital PCR in Virology: Current Applications and Future Perspectives.Molecular diagnosis & therapy · 2025Review
- Digital PCR outperforms quantitative real-time PCR for the detection and quantification of major periodontal pathobionts.Journal of oral microbiology · 2025Article
- Article
- Rapid Enzymatic Detection of Shiga-Toxin-ProducingACS infectious diseases · 2024Article
- Quantifying the Detection Sensitivity and Precision of qPCR and ddPCR Mechanisms for eDNA Samples.Ecology and evolution · 2024Article
- Generic Reporter Sets for Colorimetric Multiplex dPCR Demonstrated with 6-Plex SNP Quantification Panels.International journal of molecular sciences · 2024Article
- The Development of a Sensitive Droplet Digital Polymerase Chain Reaction Test for Quantitative Detection of Goose Astrovirus.Viruses · 2024Article
- Development of a droplet digital PCR method for detection of porcine circovirus 4.BMC veterinary research · 2023Article
- Assessing the breeding phenology of a threatened frog species using eDNA and automatic acoustic monitoring.PeerJ · 2023Article
- Development of a droplet digital PCR assay for detection of group A porcine rotavirus.Frontiers in veterinary science · 2023Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cattle are considered to be the main reservoir for Shiga toxin-producing Escherichia coli (STEC) and are often the direct or indirect source of STEC outbreaks in humans. Accurate measurement of the concentration of shed STEC in cattle feces could be a key answer to questions concerning transmission of STEC, contamination sources and efficiency of treatments at farm level. Infected animals can be identified and the contamination level quantified by real-time quantitative PCR (qPCR), which has its specific limitations. Droplet digital PCR (ddPCR) has been proposed as a method to overcome many of the drawbacks of qPCR. This end-point amplification PCR is capable of absolute quantification independent from any reference material and is less prone to PCR inhibition than qPCR. In this study, the qPCR-based protocol described by Verstraete et al. (2014) for Shiga toxin genes stx1 and stx2 and the intimin gene eae quantification was optimized for ddPCR analysis. The properties of ddPCR and qPCR using two different mastermixes (EMM: TaqMan(®) Environmental Master Mix 2.0; UMM: TaqMan(®) Universal PCR Master Mix) were evaluated, using standard curves and both artificial and natural contaminated cattle fecal samples. In addition, the susceptibility of these assays to PCR-inhibitors was investigated. Evaluation of the standard curves and both artificial and natural contaminated cattle fecal samples suggested a very good agreement between qPCR using EMM and ddPCR. Furthermore, similar sensitivities and no PCR inhibition were recorded for both assays. On the other hand, qPCR using UMM was clearly prone to PCR inhibition. In conclusion, the ddPCR technique shows potential for the accurate absolute quantification of STEC on the farms, without relying on standardized reference material.
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