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Microchip analytic system for multiplex analysis by real-time polymerase chain reaction with reagents immobilized in microreactors

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Abstract

A microchip analytic system that uses a silicon chip with immobilized in microreactor test-system for multiplex analysis of DNA by real-time polymerase chain reaction (real time PCR) was developed and optimized. We suggested the method of immobilization of PCR-components of a test-system, chose the stabilizer, and conducted the optimization of the composition of reaction mixture to achieve permanent stability of a microchip. We conducted optimization of preparation of samples using magnetic sorbent and indicated that, with 2.6 × 104 copies/ml, 60 min are necessary to obtain positive identification including time for preparation of model samples. The abilities of the created system were demonstrated on the example of microchip analysis of samples with different content of DNA, low absolute limits of detection (20 DNA copies in microreactor), and high reproducibility of the analysis.

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References

  1. Weigl, B.H., Bardell, R.L., and Cabrera, C.R., Adv. Drug Delivery Rev., 2003, vol. 55, pp. 349–377.

    Article  CAS  Google Scholar 

  2. Dittrich, P.S., Tachikawa, K., and Manz, A., Anal. Chem., 2006, vol. 78, p. e3887.

    Article  Google Scholar 

  3. Zhang, C. and Xing, D., Nucleic Acids Res., 2007, vol. 35, pp. 4223–4237.

    Article  PubMed  CAS  Google Scholar 

  4. Morrison, T., Hurley, J., Garcia, J., Yoder, K., Katz, A., Roberts, D., Cho, J., Kanigan, T., Ilyin, S.E., Horowitz, D., Dixon, J.M., and Brenan, C.J.H., Nucleic Acids Res., 2006, vol. 34, no. 18, p. 123.

    Article  Google Scholar 

  5. RF Patent no. 2 259 401 C1, 2004.

  6. US Patent no. 6 153 412, 2000.

  7. Slyadnev, M.N., Lavrova, M.V., Erkin, M.A., Kazakov, V.A., and Ganeev, A.A., Zh. Anal. Khim., 2008, vol. 63, no. 2, pp. 210–217.

    Google Scholar 

  8. Slyadnev, M.N., Lavrova, M.V., Erkin, M.A., Navolotskii, D.V., Kris’ko, A.V., and Ganeev, A.A., Nauchn. Priborostr., 2007, vol. 17, no. 3, pp. 16–24.

    CAS  Google Scholar 

  9. Paul, C.H.L., Microfluidic Lab-on-a-Chip for Chemical and Biological Analysis and Discovery. Chromatographic Science Series, Boca Raton: CRC Press, 2006, vol. 94.

    Google Scholar 

  10. Ljung, L. and Glad, T., Modeling of Dynamic Systems, New Jersey: PTR Prentice Hall, 1994.

    Google Scholar 

  11. Rebrikov, D.V., Samatov, G.A., and Trofimov, D.Yu., PTsR “v real’nom vremeni” (Real-Time PCR), Moscow: BINOM. Laboratoriya znanii, 2009.

    Google Scholar 

  12. Tichopad, A., Dilger, M., Schwarz, G., and Pfaffl, M.W., Nucleic Acid Res., 2003, vol. 31, no. 20, p. 122.

    Article  Google Scholar 

  13. Seydack, M., Springer Ser. Fluoresc., 2008, vol. 6, pp. 401–428.

    Article  CAS  Google Scholar 

  14. Tichopad, A., Dzidic, A., and Pfaffl, M., Biotechnol. Lett., 2002, vol. 24, pp. 2053–2056.

    Article  CAS  Google Scholar 

  15. McPherson, M.J. and Mooler, S.G., PCR, Oxford: BIOS Scientific Publ. LTD, 2000, p. 276.

    Google Scholar 

  16. Sachse, K. and Frey, J., Methods Mol. Biol., Totowa, New Jersey: Humana Press Inc., 2002, vol. 216.

    Google Scholar 

  17. Pelt-Verkuil, E., Belkum, A., and Hays, J.P., Principles and Technical Aspects of PCR Amplification, Netherlands: Springer, 2008.

    Book  Google Scholar 

  18. Olsvik, O., Popovic, T., Skjerve, E., Cudjoe, K.S., Homes, E., Ugelstad, J., and Uhlen, M., Clinical Microbiol. Rev., 1994, vol. 7, no. 1, pp. 43–54.

    CAS  Google Scholar 

  19. Berensmeier, S., Appl. Microbiol. Biotechnol., 2006, vol. 73, pp. 495–504.

    Article  PubMed  CAS  Google Scholar 

  20. Gray, J.P. and Herwig, R.P., Appl. Environ. Microbiol., 1996, no. 11, pp. 4049–4059.

  21. LeBreton, M., Morton, P., Larade, K., Harland, B., Clair, T., and Campbell, D., Hydrobiologia, 2000, vol. 438, no. 1, pp. 91–97.

    Article  CAS  Google Scholar 

  22. Kuske, C.R., Banton, K.L., Adorada, D.L., Stark, P.C., Hill, K.K., and Jackson, P.J., Appl. Environ. Microbiol., 1998, vol. 64, no. 7, pp. 2463–2472.

    PubMed  CAS  Google Scholar 

  23. Saikaly, P.E., Barlaz, M.A., Francis, L., and de Los Reyes, F.L., III, Appl. Environ. Microbiol., 2007, vol. 73, no. 20, pp. 6557–6565.

    Article  PubMed  CAS  Google Scholar 

  24. Trapmann, S., Catalani, P., Conneely, P., Contreras, M., Corbisier, P., Gancberg, D., and Gloria, S., Linsinger, T., Zeleny, R., and Schimmel, K., The Certification of Reference Materials of Dry-Mixed Maize Powder with different Mass Fractions of Bt-176 Maize, Luxembourg: Official Publ. Eur. Comm., 2004.

    Google Scholar 

  25. Consolandi, C., Severgnini, M., Frosini, A., Caramenti, G., De Fazio, M., Ferrara, F., Zocco, A., Fischetti, A., Palmieri, M., and De Bellis, G., Anal. Biochem., 2006, vol. 353, no. 2, pp. 191–197.

    Article  PubMed  CAS  Google Scholar 

  26. Lee, J.G., Cheong, K.H., Huh, N., Kim, S., Choi, J.W., and Ko, S., Lab. Chip, 2006, vol. 6, pp. 886–895.

    Article  PubMed  CAS  Google Scholar 

  27. Arumuganathan, K. and Earle, E.D., Plant Mol. Biol. Report., 1991, vol. 9, pp. 208–218.

    Article  CAS  Google Scholar 

  28. Slyadnev, M.H., Lavrova, M.V., Erkin, M.A., Navolotskii, D.V., Kris’ko, A.V., and Ganeev, A.A., Nauchn. Priborostr., 2007, vol. 17, no. 3, pp. 25–30.

    CAS  Google Scholar 

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Authors and Affiliations

  1. Lumex Group of Companies, St. Petersburg, 192029, Russia

    D. V. Navolotskii, A. V. Perchik, A. A. Ganeev & M. N. Slyadnev

  2. St. Petersburg State University Institute of Chemistry, St. Petersburg, 192029, Russia

    D. V. Navolotskii, I. A. Mark’yanov, A. A. Ganeev & M. N. Slyadnev

Authors
  1. D. V. Navolotskii
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  2. A. V. Perchik
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  3. I. A. Mark’yanov
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  4. A. A. Ganeev
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  5. M. N. Slyadnev
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Correspondence to A. V. Perchik.

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Original Russian Text © D.V. Navolotskii, A.V. Perchik, I.A. Mark’yanov, A.A. Ganeev, M.N. Slyadnev, 2011, published in Prikladnaya Biokhimiya i Mikrobiologiya, 2011, Vol. 47, No. 2, pp. 241–248.

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Navolotskii, D.V., Perchik, A.V., Mark’yanov, I.A. et al. Microchip analytic system for multiplex analysis by real-time polymerase chain reaction with reagents immobilized in microreactors. Appl Biochem Microbiol 47, 221–227 (2011). https://doi.org/10.1134/S0003683811020141

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  • DOI: https://doi.org/10.1134/S0003683811020141

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