Skip to main content
SpringerLink
Log in

Single-Base Extension and ELISA-Based Approach for Single-Nucleotide Polymorphisms Genotyping

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

Single-nucleotide polymorphisms (SNPs) emerge as a fundamental tool in personalized medicine due to their association with drug responses or disease predisposition. Single-base extension (SBE) is a common method for characterizing known SNPs, but involves complicated procedures or requires costly analytical instruments. Here, we describe a novel SNP genotyping based on SBE and enzyme-linked immunosorbent assay (ELISA). During the SBE, the 5′ end fluorescein isothiocyanate-labeled allele-specific primer will extend with biotinylated dideoxynucleotides which are complementary to the SNP sites. The extension product will then be captured by streptavidin-coated nanoparticle and develop blue color in the ELISA assay. We validated this method by detecting SNPs for TP53 gene codon 273 from 68 individuals and the data were 100% in concordant with DNA sequencing. Thus, SBE and ELISA-based SNPs assay is a simple and accurate method for SNP genotyping.

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

Fig. 1

Similar content being viewed by others

References

  1. Sachidanandam, R., Weissman, D., Schmidt, S. C., Kakol, J. M., Stein, L. D., Marth, G., et al. (2001). Nature, 409, 928–933.

    Article  CAS  Google Scholar 

  2. Brahmachari, S. K. (2001). Applied Biochemistry and Biotechnology, 96, 9–12.

    Article  CAS  Google Scholar 

  3. Kwok, P. Y., Carlson, C., Yager, T. D., Ankener, W., & Nickerson, D. A. (1994). Genomics, 23, 38–144.

    Article  Google Scholar 

  4. Wang, D. G., Fan, J. B., Siao, C. J., Berno, A., Young, P., Sapolsky, R., et al. (1998). Science, 280, 1077–1082.

    Article  CAS  Google Scholar 

  5. Wenz, H. M., Robertson, J. M., Menchen, S., Oaks, F., Demorest, D. M., Scheibler, D., et al. (1998). Genome Research, 8, 69–80.

    CAS  Google Scholar 

  6. Boon, E. M., Ceres, D. M., Drummond, T. G., Hill, M. G., & Barton, J. K. (2000). Nature Biotechnology, 18, 1096–1100.

    Article  CAS  Google Scholar 

  7. Haff, L. A., & Smirnov, I. P. (1997). Nucleic Acids Research, 25, 3749–3750.

    Article  CAS  Google Scholar 

  8. Landegren, U., Kaiser, R., Sanders, J., & Hood, L. (1988). Science, 241, 1077–1080.

    Article  CAS  Google Scholar 

  9. Mhlanga, M. M., & Malmberg, L. (2001). Methods, 25, 463–471.

    Article  CAS  Google Scholar 

  10. Syvänen, A. C., Aalto-Steälä, K., Harju, L., Kontula, K., & Söderlund, H. (1990). Genomics, 8, 684–692.

    Article  Google Scholar 

  11. Raitio, M., Lindroos, K., Laukkanen, M., Pastinen, T., Sistonen, P., Sajantila, A., et al. (2001). Genome Research, 11, 471–482.

    Article  CAS  Google Scholar 

  12. Hirschhorn, J. N., Sklar, P., Lindblad-Toh, K., Lim, Y. M., Ruiz-Gutierrez, M., Bolk, S., et al. (2000). Proceedings of the National Academy of Sciences of the United States of America, 97, 12164–12169.

    Article  CAS  Google Scholar 

  13. Tong, A. K., & Ju, J. (2002). Nucleic Acids Research, 30, e19.

    Article  Google Scholar 

  14. Takatsu, K., Yokomaku, T., Kurata, S., & Kanagawa, T. (2004). Nucleic Acids Research, 32, e156.

    Article  Google Scholar 

Download references

Acknowledgments

We thank Dr. Yingqun Wang for suggestion and editorial assistance. This work was supported by funding from Science and Technology Bureau of Heilongjiang Province (grant no. LC06C25).

Competing Interests

The authors declare no competing interests.

Author information

Authors and Affiliations

  1. Department of Anatomy, Mudanjiang Medical College, Mudanjiang, China

    Guibo Liu & Wei Zhao

  2. Department of Pathology, Mudanjiang Medical College, Mudanjiang, China

    Yongxia Cheng & Zaishun Jin

  3. Hongbo Shan, ADICON Clinical Lab, Hangzhou, China

    Hongbo Shan

  4. Gaolian XU, Ustar Biotechnology.Ltd, Hangzhou, China

    Gaolian Xu

Authors
  1. Guibo Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yongxia Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wei Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zaishun Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hongbo Shan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Gaolian Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zaishun Jin.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liu, G., Cheng, Y., Zhao, W. et al. Single-Base Extension and ELISA-Based Approach for Single-Nucleotide Polymorphisms Genotyping. Appl Biochem Biotechnol 163, 573–576 (2011). https://doi.org/10.1007/s12010-010-9063-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-010-9063-4

Keywords

Search

Navigation