Exploring the influence of vitamin C concentrations on the dynamics of RT-PCR assay reactions

Sharafi et al., Scientific Reports, doi:10.1038/s41598-025-91154-1, Jul 2025

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https://c19early.org/sharafi.html

Vitamin C for COVID-19

6th treatment shown to reduce risk in September 2020, now with p = 0.00000002 from 75 studies, recognized in 22 countries.

Lower risk for mortality, ICU, hospitalization, and recovery.

No treatment is 100% effective. Protocols combine treatments.

6,000+ studies for 175 treatments. c19early.org

In vitro study showing that vitamin C may interfere with COVID-19 RT-PCR diagnostics by altering reaction kinetics. High vitamin C concentrations (50-100 mg/ml) significantly increased Ct values in samples with high viral load while decreasing values in samples with low viral load. The concentrations are supraphysiologic - 50-100 mg ml⁻¹ far exceeds serum levels achieved by oral or standard IV dosing, so the finding is most relevant to intranasal or direct-sample contamination scenarios. In clinical treatment studies to date, benefit is seen only for serious outcomes, with no significant effect seen for viral outcomes.

17 preclinical studies support the efficacy of vitamin C for COVID-19:

8 In Silico studies1-8

8 In Vitro studies1,7,9-14

1 In Vivo animal study12

Vitamin C has been identified by the European Food Safety Authority (EFSA) as having sufficient evidence for a causal relationship between intake and optimal immune system function15-17. Vitamin C plays a key role in the immune system, supporting the production and function of leukocytes, or white blood cells, which defend against infection and disease, including the production of lymphocytes, which make antibodies, and enhancing phagocytosis, the process by which immune system cells ingest and destroy viruses and infected cells. Vitamin C is an antioxidant, protecting cells from damage caused by free radicals. Vitamin C inhibits SARS-CoV-2 3CL^pro7,11, inhibits SARS-CoV-2 infection by reducing ACE2 levels in a dose-dependent manner12, and may limit COVID-19 induced cardiac damage by acting as an antioxidant and potentially reducing the reactive oxygen species (ROS) production induced by the spike protein that contributes to the activation of profibrotic pathways9. Vitamin C reduces inflammation, oxidative stress, and NETosis, supporting immune function and vascular protection18. Intracellular levels of vitamin C decline during COVID-19 hospitalization suggesting ongoing utilization and depletion of vitamin C19. Threonic acid, a metabolite of vitamin C, is lower in mild and severe cases, consistent with increased need for and metabolization of vitamin C with moderate infection, but more limited ability to produce threonic acid in severe infection due to depletion or existing lower levels of vitamin C20. Symptomatic COVID-19 is associated with a lower frequency of natural killer (NK) cells, and vitamin C has been shown to improve NK cell numbers and functioning21,22.

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1. Najimi et al., Phytochemical Inhibitors of SARS‐CoV‐2 Entry: Targeting the ACE2‐RBD Interaction with l‐Tartaric Acid, l‐Ascorbic Acid, and Curcuma longa Extract, ChemistrySelect, doi:10.1002/slct.202406035.wiley.com, doi.org.

2. Rajamanickam et al., Exploring the Potential of Siddha Formulation MilagaiKudineer-Derived Phytotherapeutics Against SARS-CoV-2: An In-Silico Investigation for Antiviral Intervention, Journal of Pharmacy and Pharmacology Research, doi:10.26502/fjppr.0105.fortunejournals.com, doi.org.

3. Agamah et al., Network-based multi-omics-disease-drug associations reveal drug repurposing candidates for COVID-19 disease phases, ScienceOpen, doi:10.58647/DRUGARXIV.PR000010.v1.scienceopen.com, doi.org.

4. Morales-Bayuelo et al., New findings on ligand series used as SARS-CoV-2 virus inhibitors within the frameworks of molecular docking, molecular quantum similarity and chemical reactivity indices, F1000Research, doi:10.12688/f1000research.123550.3.f1000research.com, doi.org.

5. Alkafaas et al., A study on the effect of natural products against the transmission of B.1.1.529 Omicron, Virology Journal, doi:10.1186/s12985-023-02160-6.biomedcentral.com, doi.org.

6. Pandya et al., Unravelling Vitamin B12 as a potential inhibitor against SARS-CoV-2: A computational approach, Informatics in Medicine Unlocked, doi:10.1016/j.imu.2022.100951.sciencedirect.com, doi.org.

7. Malla et al., Vitamin C inhibits SARS coronavirus-2 main protease essential for viral replication, bioRxiv, doi:10.1101/2021.05.02.442358.biorxiv.org, doi.org.

8. Kumar et al., In silico virtual screening-based study of nutraceuticals predicts the therapeutic potentials of folic acid and its derivatives against COVID-19, VirusDisease, doi:10.1007/s13337-020-00643-6.springer.com, doi.org.

9. Van Tin et al., Spike Protein of SARS-CoV-2 Activates Cardiac Fibrogenesis through NLRP3 Inflammasomes and NF-κB Signaling, Cells, doi:10.3390/cells13161331.mdpi.com, doi.org.

10. Moatasim et al., Potent Antiviral Activity of Vitamin B12 against Severe Acute Respiratory Syndrome Coronavirus 2, Middle East Respiratory Syndrome Coronavirus, and Human Coronavirus 229E, Microorganisms, doi:10.3390/microorganisms11112777.mdpi.com, doi.org.

11. Đukić et al., Inhibition of SARS-CoV-2 Mpro with Vitamin C, L-Arginine and a Vitamin C/L-Arginine Combination, Frontiers in Bioscience-Landmark, doi:10.31083/j.fbl2801008.imrpress.com, doi.org.

12. Zuo et al., Vitamin C promotes ACE2 degradation and protects against SARS‐CoV‐2 infection, EMBO reports, doi:10.15252/embr.202256374.wiley.com, doi.org.

13. Hajdrik et al., In Vitro Determination of Inhibitory Effects of Humic Substances Complexing Zn and Se on SARS-CoV-2 Virus Replication, Foods, doi:10.3390/foods11050694.mdpi.com, doi.org.

14. Goc et al., Inhibitory effects of specific combination of natural compounds against SARS-CoV-2 and its Alpha, Beta, Gamma, Delta, Kappa, and Mu variants, European Journal of Microbiology and Immunology, doi:10.1556/1886.2021.00022.akjournals.com, doi.org.

15. Galmés et al., Suboptimal Consumption of Relevant Immune System Micronutrients Is Associated with a Worse Impact of COVID-19 in Spanish Populations, Nutrients, doi:10.3390/nu14112254.mdpi.com, doi.org.

16. Galmés (B) et al., Current State of Evidence: Influence of Nutritional and Nutrigenetic Factors on Immunity in the COVID-19 Pandemic Framework, Nutrients, doi:10.3390/nu12092738.mdpi.com, doi.org.

17. EFSA, Scientific Opinion on the substantiation of health claims related to vitamin C and protection of DNA, proteins and lipids from oxidative damage (ID 129, 138, 143, 148), antioxidant function of lutein (ID 146), maintenance of vision (ID 141, 142), collagen formation (ID 130, 131, 136, 137, 149), function of the nervous system (ID 133), function of the immune system (ID 134), function of the immune system during and after extreme physical exercise (ID 144), non-haem iron absorption (ID 132, 147), energy-yielding metabolism (ID 135), and relief in case of irritation in the upper respiratory tract (ID 1714, 1715) pursuant to Article 13(1) of Regulation (EC) No 1924/2006, EFSA Journal, doi:10.2903/j.efsa.2009.1226.europa.eu, doi.org.

18. Xie et al., The role of reactive oxygen species in severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) infection-induced cell death, Cellular & Molecular Biology Letters, doi:10.1186/s11658-024-00659-6.biomedcentral.com, doi.org.

19. Boerenkamp et al., Low Levels of Serum and Intracellular Vitamin C in Hospitalized COVID-19 Patients, Nutrients, doi:10.3390/nu15163653.mdpi.com, doi.org.

20. Albóniga et al., Differential abundance of lipids and metabolites related to SARS-CoV-2 infection and susceptibility, Scientific Reports, doi:10.1038/s41598-023-40999-5.nature.com, doi.org.

21. Graydon et al., High baseline frequencies of natural killer cells are associated with asymptomatic SARS-CoV-2 infection, Current Research in Immunology, doi:10.1016/j.crimmu.2023.100064.sciencedirect.com, doi.org.

22. Vojdani et al., In vivo effect of ascorbic acid on enhancement of human natural killer cell activity, Nutrition Research, doi:10.1016/S0271-5317(05)80799-7.sciencedirect.com, doi.org.

Sharafi et al., 29 Jul 2025, peer-reviewed, 6 authors. Contact: psharafi@etu.edu.tr.

This Paper Vitamin C All

Exploring the influence of vitamin C concentrations on the dynamics of RT-PCR assay reactions

Parisa Sharafi, Mesut Akyol, Ebrar Gultekin, Rohat Sakar, N Yasemin Ardicoglu Akisin, J Sedef Gocmen

Scientific Reports, doi:10.1038/s41598-025-91154-1

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that caused the COVID-19 pandemic to break out touched off a global health catastrophe characterized by various degrees of disease severity among those who were afflicted. Many strategies, such as vitamin C administration, have been investigated to reduce COVID-19 symptoms. Although the exact processes by which vitamin C affects COVID-19 remain unclear, noticeable changes in PCR test results were noted in our laboratory settings. This study uses PCR analysis to investigate the effects of varying vitamin C dosages and durations on COVID-19 test results. PCR cycle threshold (Ct) values were used to categorize nasopharyngeal tissues from 98 patients (Ct < 30, Ct ≥ 30, negative). Vitamin C was applied at different concentrations (0, 1, 10, 50, and 100 mg/ml), and PCR analyses were carried out at 1, 10, 24, and 48 h marks after the vitamin was applied. Particularly in samples with lower Ct values, the data showed significant changes in the reaction graphs and metrics with increasing vitamin C concentration. Higher concentrations of vitamin C were correlated with diminished metrics, occasionally leading to negative results for samples with Ct ≥ 30 values. Notably, samples that showed no discernible viral loads had different pictorial representations. These results raise questions regarding the reliability of PCR results in the presence of vitamin C intake and have implications for COVID-19 diagnosis. In light of the current pandemic, more studies are necessary to confirm and expand these findings and provide a critical understanding of clinical procedures and the interpretation of test results.

administered 0 and 1 mg/ml vitamin C were higher than those for samples administered 50 and 100 mg/ml vitamin C. Additionally, the median Ct values for 50 and 100 mg/ml vitamin C doses were very low, which was attributed to graphical image disruption in the PCR test. In the 10th hour for samples exhibiting a negative viral load, it was noted that at least one dose yielded distinct effects compared to others (χ 2 = 13.063; p = 0.011). Upon investigating the cause of this difference, a significant difference was observed between the 0 and 10 mg/ml doses (Z = 3.213; p = 0.013), whereas the other vitamin C doses did not exhibit statistical significance (p > 0.05). The median Ct value of samples administered 0 mg/ml vitamin C was higher than that of samples administered 10 mg/ml vitamin C. The PCR test reaction was affected by the deterioration of the graphic images, which was the reason for this discrepancy. In the 24th hour, for samples with a high viral load (Ct < 30), it was observed that at least one dose had a distinct effect compared to the others (χ 2 = 64.160; p < 0.001). Upon investigating the reason for this difference, statistically significant differences emerged between low doses (0, 1, and 10 mg/ml) and high doses (50 and 100 mg/ml) of Vitamin C (P < 0.05), with no significant difference between low and high doses (P > 0.05). The median Ct values of samples treated with low doses of Vitamin C were notably lower than those of samples treated with high..

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