Skip to main content

Advertisement

SpringerLink
Log in

A Turn Off Fluorescence Probe Based on Carbon Dots for Highly Sensitive Detection of BRCA1 Gene in Real Samples and Cellular Imaging

  • Original Article
  • Published:
Journal of Fluorescence Aims and scope Submit manuscript

Abstract

In this research, DNA-modified carbon dots (CDs) were exploited to construct a fluorescence assay for breast cancer genes (BRCA1, a potential marker for cancer diagnosis) detection. For this purpose, water-soluble synthesized CDs were functionalized with 19 mer-modified oligonucleotides (capture probe). By adding the DNA target, the specific binding between the DNA probe and DNA target causes fluorescence quenching. The assay displayed a fine capability of sensing the BRCA1 gene with a linear range (R2 = 0.9918) of 36 attomolar (aM) to 532 femtomolar (fM) and a detection limit of 2 attomolar. This homogeneous process does not need additional separation and washing steps of un-hybridized DNA. To assess the selectivity, the prepared biosensor responses were evaluated in solutions containing single-base mismatched DNA sequences, three-base mismatched DNA sequences, or non-complementary DNA sequences, separately. To demonstrate the practical application of the designed biosensor, the extracted DNA from blood samples of breast cancer patients was utilized as real samples. When the CDs-DNA bioassay was exploited in the imaging of MCF-7 cancer cells, strong fluorescence emission was observed. After incubation times, both the cells’ size and shape remained unchanged. The results validated that the CDs are an extremely great bioimaging candidate in disease diagnosis, biomedicine investigation, and managing cancer diseases.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Scheme 1
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Data Availability

All data generated or analyzed during this study are included in this published article.

Code Availability

Not Applicable.

References

  1. Smith SJ, Nemr CR, Kelley SO (2017) Chemistry-driven approaches for ultrasensitive nucleic acid detection. J Am Chem Soc 139(3):1020–1028

    Article  CAS  Google Scholar 

  2. Vikrant K, Bhardwaj N, Bhardwaj SK, Kim KH, Deep A (2019) Nanomaterials as efficient platforms for sensing DNA. Biomaterials 214:119215

    Article  CAS  Google Scholar 

  3. Abdulá Rasheed P (2015) A highly sensitive DNA sensor for attomolar detection of the BRCA1 gene: signal amplification with gold nanoparticle clusters. Analyst 140(8):2713–2718

    Article  Google Scholar 

  4. Rasheed PA, Sandhyarani N (2015) Attomolar detection of BRCA1 gene based on gold nanoparticle assisted signal amplification. Biosens Bioelectron 65:333–340

    Article  Google Scholar 

  5. Borghei YS, Hosseini M, Ganjali MR, Ju HA (2019) unique FRET approach toward detection of single-base mismatch DNA in BRCA1 gene. Mater Sci Eng C 97:406–411

    Article  CAS  Google Scholar 

  6. Borghei YS, Hosseini M, Ganjali MR, Hosseinkhani S (2018) A novel BRCA1 gene deletion detection in human breast carcinoma MCF-7 cells through FRET between quantum dots and silver nanoclusters. J Pharm Biomed Anal 152:81–88

    Article  CAS  Google Scholar 

  7. Li CZ, Karadeniz H, Canavar E, Erdem A (2012) Electrochemical sensing of label free DNA hybridization related to breast cancer 1 gene at disposable sensor platforms modified with single walled carbon nanotubes. Electrochim Acta 82:137–142

    Article  CAS  Google Scholar 

  8. Rasheed PA, Sandhyarani N (2014) Graphene-DNA electrochemical sensor for the sensitive detection of BRCA1 gene. Sens Actuators B Chem 204:777–782

    Article  CAS  Google Scholar 

  9. Rasheed PA, Radhakrishnan T, Shihabudeen PK, Sandhyarani N (2016) Reduced graphene oxide-yttria nanocomposite modified electrode for enhancing the sensitivity of electrochemical genosensor. Biosens Bioelectron 83:361–367

    Article  CAS  Google Scholar 

  10. García-Mendiola T, Elosegui CG, Bravo I, Pariente F, Jacobo-Martin A, Navio C, Rodriguez I, Wannemacher R, Lorenzo E (2019) Fluorescent C-NanoDots for rapid detection of BRCA1, CFTR and MRP3 gene mutations. Microchim Acta 186(5):293

    Article  Google Scholar 

  11. Wang L, Bi Y, Hou J, Li H, Xu Y, Wang B, Ding H, Ding L (2016) Facile, green and clean one-step synthesis of carbon dots from wool: application as a sensor for glyphosate detection based on the inner filter effect. Talanta 160:268–275

    Article  CAS  Google Scholar 

  12. Liang Q, Ma W, Shi Y, Li Z, Yang X (2013) Easy synthesis of highly fluorescent carbon quantum dots from gelatin and their luminescent properties and applications. Carbon 60:421–428

    Article  CAS  Google Scholar 

  13. Li LS, Jiao XY, Zhang Y, Cheng C, Huang K, Xu L (2018) Green synthesis of fluorescent carbon dots from Hongcaitai for selective detection of hypochlorite and mercuric ions and cell imaging. Sens Actuators B Chem 263:426–435

    Article  CAS  Google Scholar 

  14. Mandal D, Mishra S, Singh RK (2018) Green Synthesized Nanoparticles as Potential Nanosensors. Environmental, Chemical and Medical Sensors, Publisher: Springer Singapore, Print ISBN: 978-981-10-7750-0, Electronic ISBN: 978-981-10-7751-7

  15. Ganiga M, Cyriac J (2016) FRET based ammonia sensor using carbon dots. Sens Actuators B Chem 225:522–528

    Article  CAS  Google Scholar 

  16. Mohammadi S, Salimi A (2018) Fluorometric determination of microRNA-155 in cancer cells based on carbon dots and MnO2 nanosheets as a donor-acceptor pair. Microchim Acta 185(8):372

    Article  Google Scholar 

  17. Mohammadi S, Salimi A, Hamd-Ghadareh S, Fathi F, Soleimani F (2018) A FRET immunosensor for sensitive detection of CA 15 – 3 tumor marker in human serum sample and breast cancer cells using antibody functionalized luminescent carbon-dots and AuNPs-dendrimer aptamer as donor-acceptor pair. Anal biochem 557:18–26

    Article  CAS  Google Scholar 

  18. Lorigooini Z, Koravand M, Haddadi H, Rafieian-Kopaei M, Shirmardi HA, Hosseini Z (2019) A review of botany, phytochemical and pharmacological properties of Ferulago angulate. Toxin Rev 38(1):13–20

    Article  CAS  Google Scholar 

  19. Pirsaheb M, Mohammadi S, Salimi A, Payandeh M (2019) Functionalized fluorescent carbon nanostructures for targeted imaging of cancer cells: a review. Microchim Acta 186(4):231

    Article  Google Scholar 

  20. Xia Y, Wang L, Li J, Chen X, Lan J, Yan A, Yan L, Yang S, Yang H, Chen J (2018) A ratiometric fluorescent bioprobe based on carbon dots and acridone derivate for signal amplification detection exosomal microRNA. Anal Chem 90(15):8969–8976

    Article  CAS  Google Scholar 

  21. Chacon-Cortes D, Griffiths LR (2014) Methods for extracting genomic DNA from whole blood samples: current perspectives. J Biorepository Sci Appl Med 2014(2):1–9

    Google Scholar 

  22. Hosseinkhani Z, Sadeghalvad M, Norooznezhad F, Khodarahmi R, Fazilati M, Mahnam A, Fattahi A, Mansouri K (2018) The effect of CYP2C9* 2, CYP2C9* 3, and VKORC1-1639 G > A polymorphism in patients under warfarin therapy in city of Kermanshah. Res Pharm Sci 13(4):377

    Article  Google Scholar 

  23. Couto MCM, Sudre AP, Lima MF, Bomfim TCB (2013) Comparison of techniques for DNA extraction and agarose gel staining of DNA fragments using samples of Cryptosporidium. Vet Med 58(10):535–542

    Article  Google Scholar 

  24. Ni P, Xie J, Chen C, Jiang Y, Lu Y, Hu X (2019) Fluorometric determination of the activity of alkaline phosphatase and its inhibitors based on ascorbic acid-induced aggregation of carbon dots. Microchim Acta 186(3):202

    Article  Google Scholar 

  25. Barati A, Shamsipur M, Abdollahi H (2016) Carbon dots with strong excitation-dependent fluorescence changes towards pH. Application as nanosensors for a broad range of pH. Anal Chim Acta 931:25–33

    Article  CAS  Google Scholar 

  26. Wu P, Li W, Wu Q, Liu Y, Liu S (2017) Hydrothermal synthesis of nitrogen-doped carbon quantum dots from microcrystalline cellulose for the detection of Fe 3+ ions in an acidic environment. RSC adv 7(70):44144–44153

    Article  CAS  Google Scholar 

  27. Shen J, Shang S, Chen X, Wang D, Cai Y (2017) Facile synthesis of fluorescence carbon dots from sweet potato for Fe3 + sensing and cell imaging. Mater Sci Eng C 76:856–864

    Article  CAS  Google Scholar 

  28. Qaddare SH, Salimi A (2017) Amplified fluorescent sensing of DNA using luminescent carbon dots and AuNPs/GO as a sensing platform: a novel coupling of FRET and DNA hybridization for homogeneous HIV-1 gene detection at femtomolar level. Biosens Bioelectron 89:773–780

    Article  CAS  Google Scholar 

  29. Zu F, Yan F, Bai Z, Xu J, Wang Y, Huang Y, Zhou X (2017) The quenching of the fluorescence of carbon dots: A review on mechanisms and applications. Microchim Acta 184:1899–1914

    Article  CAS  Google Scholar 

  30. Senol AM, Bozkurt E (2020) Facile green and one-pot synthesis of seville orange derived carbon dots as a fluorescent sensor for Fe3+ ions. Microchem J 159:105357

    Article  CAS  Google Scholar 

  31. Park JY, Park SM (2009) DNA Hybridization Sensors Based on Electrochemical Impedance Spectroscopy as a Detection Tool. Sensors 9:9513–9532

    Article  CAS  Google Scholar 

  32. Pirsaheb M, Mohammadi S, Salimi A (2019) Current advances of carbon dots based biosensors for tumor marker detection, cancer cells analysis and bioimaging. TRAC-Trend Anal Chem 115:83–99

    Article  CAS  Google Scholar 

  33. Tuerhong M, Yang X, Xue-Bo Y (2017) Review on Carbon Dots and Their Applications. Chin J Anal Chem 45(1):139–150

    Article  Google Scholar 

  34. Yang B, Zhang S, Fang X, Kong J (2019) Double signal amplification strategy for ultrasensitive electrochemical biosensor based on nuclease and quantum dot-DNA nanocomposites in the detection of breast cancer 1 gene mutation. Biosens Bioelectron 142:111544

    Article  CAS  Google Scholar 

  35. Zhang H, Wang L, Jiang W (2011) Label free DNA detection based on gold nanoparticles quenching fluorescence of Rhodamine B. Talanta 85(1):725–729

    Article  Google Scholar 

Download references

Acknowledgements

The authors gratefully acknowledge the Research Council of Kermanshah University of Medical Sciences (Grant Number: 97041) for the financial support.

Funding

The authors gratefully acknowledge the Research Council of Kermanshah University of Medical Sciences (Grant Number: 97041) for the financial support.

Author information

Authors and Affiliations

  1. Research Center for Environmental Determinants of Health (RCEDH), Kermanshah University of Medical Sciences, 67198-51351, Kermanshah, Iran

    Meghdad Pirsaheb & Somayeh Mohammadi

  2. Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran

    Reza Khodarahmi, Zohreh Hoseinkhani & Kamran Mansouri

  3. Regenerative Medicine and BMT, Kermanshah University of Medical Sciences, 67148-69914, Kermanshah, Iran

    Mehrdad Payandeh

Authors
  1. Meghdad Pirsaheb
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Somayeh Mohammadi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Reza Khodarahmi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zohreh Hoseinkhani
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kamran Mansouri
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mehrdad Payandeh
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

MP helped through supervision, project administration, funding acquisition, SM performed investigation, data curation, conceptualization, writing - original draft and editing, RKh contributed to supervision, Editing. ZH performed cell culture experiments and a cellular cytotoxicity test. KM helped by supervision. MP Supplied the blood samples from breast cancer patients.

Corresponding author

Correspondence to Somayeh Mohammadi.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest in the publication of this article.

Ethics approval/declarations

Not applicable.

Consent to Participate

Not applicable.

Consent for Publication

Not applicable.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Pirsaheb, M., Mohammadi, S., Khodarahmi, R. et al. A Turn Off Fluorescence Probe Based on Carbon Dots for Highly Sensitive Detection of BRCA1 Gene in Real Samples and Cellular Imaging. J Fluoresc 32, 1733–1741 (2022). https://doi.org/10.1007/s10895-022-02954-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10895-022-02954-x

Keywords

Search

Navigation