ArticleAnalytical and bioanalytical chemistry2018
Inhibition mechanisms of hemoglobin, immunoglobulin G, and whole blood in digital and real-time PCR.
Article in Analytical and bioanalytical chemistry, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 108 papers, 1 of them a synthesis that pooled it.
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Who cites it
108 citing papers in PubMed, 1 synthesis or guideline pooled it, 219 citations in OpenAlex.
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- Urine Cell-Free DNA in the Diagnosis of Intraductal Papillary Mucinous Neoplasms: A Prospective Observational Study.Annals of surgical oncology · 2026Observational
- Amplification of RNA for identification of Zika and HCV in whole blood.Science advances · 2026Article
- Rapid, low-cost, and portable molecular detection of HIV-1 subtype B and HCV 1a directly from blood.Microbiology spectrum · 2026Article
- Detecting swine influenza A virus during fattening and at slaughter: implications for monitoring strategies in fattening herds.Porcine health management · 2026Article
- A Tailored Workflow for Circular RNA Enrichment and Analysis from Human Whole Blood.Biomolecules · 2026Article
- Molecular Characterization and Prevalence ofPathogens (Basel, Switzerland) · 2026Article
- Detrimental effect of metal contaminants on forensic DNA analysis and strategies for mitigation.International journal of legal medicine · 2026Review
- Development of a Highly Specific RPA/CRISPR-Cas13a Assay for Detection ofCurrent issues in molecular biology · 2026Article
- Rapid and reliable diagnosis of mucormycosis using colorimetric loop-mediated isothermal amplification.Journal of clinical microbiology · 2026Article
- Advances of Digital Detection for Foodborne Pathogens.Foods (Basel, Switzerland) · 2026Review
- Mock Samples That Mimic Human Cervicovaginal Samples to Accelerate the Development and Evaluation of Assays for High-Risk HPV for Cervical Cancer Screening.Journal of medical virology · 2026Article
- Comparative Evaluation of Rapid Nucleic Acids Extraction Methods for Biosensor-Based Point-of-Care Solutions.Biosensors · 2026Article
- Systematic review and meta-analysis on the diagnostic accuracy of various detection methods for porcine reproductive and respiratory syndrome virus.Porcine health management · 2026Article
- Comparative Evaluation of Automated Nucleic Acid Extraction Systems for DNA and RNA Viral Target.Pathogens (Basel, Switzerland) · 2026Article
- teamNGS Balances Sensitivity for Viruses with Comprehensive Microbial Detection in Clinical Specimens.Microorganisms · 2025Article
- Comparison of boiling versus magnetic bead techniques in nucleic acid extraction for human papillomavirus detection: evidence based 17,179 cases.Virology journal · 2025Article
- Towards practical point-of-care quick, ubiquitous, integrated, cost-efficient molecular diagnostic Kit (QUICK) PCR for future pandemic response.Microsystems & nanoengineering · 2025Review
- Development and diagnostic performance of a sensitive multiplex pneumonia pathogen identification assay for emergency department patients with pneumonia.Microbiology spectrum · 2025Article
48 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
8 authors at 5 institutions in 3 countries.
Funding
Abstract
Blood samples are widely used for PCR-based DNA analysis in fields such as diagnosis of infectious diseases, cancer diagnostics, and forensic genetics. In this study, the mechanisms behind blood-induced PCR inhibition were evaluated by use of whole blood as well as known PCR-inhibitory molecules in both digital PCR and real-time PCR. Also, electrophoretic mobility shift assay was applied to investigate interactions between inhibitory proteins and DNA, and isothermal titration calorimetry was used to directly measure effects on DNA polymerase activity. Whole blood caused a decrease in the number of positive digital PCR reactions, lowered amplification efficiency, and caused severe quenching of the fluorescence of the passive reference dye 6-carboxy-X-rhodamine as well as the double-stranded DNA binding dye EvaGreen. Immunoglobulin G was found to bind to single-stranded genomic DNA, leading to increased quantification cycle values. Hemoglobin affected the DNA polymerase activity and thus lowered the amplification efficiency. Hemoglobin and hematin were shown to be the molecules in blood responsible for the fluorescence quenching. In conclusion, hemoglobin and immunoglobulin G are the two major PCR inhibitors in blood, where the first affects amplification through a direct effect on the DNA polymerase activity and quenches the fluorescence of free dye molecules, and the latter binds to single-stranded genomic DNA, hindering DNA polymerization in the first few PCR cycles. Graphical abstract PCR inhibition mechanisms of hemoglobin and immunoglobulin G (IgG). Cq quantification cycle, dsDNA double-stranded DNA, ssDNA single-stranded DNA.
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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.