Abstract
Economical, religious, and health reasons demand an accurate control of food in order to protect consumers from falsely labeled products. Meat in particular is easily susceptible to fraudulent labeling, mainly through contamination with species of lower value. New methods and protocols for rapid, sensitive and reliable identification of extraneous species in food are therefore required. The miniaturization and optimization of analytical methodologies are powerful tools in this direction, especially when connected to Lab-on-a-chip (LOC) microdevices. LOCs possess many advantages, such as the reduction of the analysis cost, the possibility to save time and labor, the easiness of use and not last, the possibility to bring a complex technique out of the laboratory. Here we present a new concept for the food quality control, i.e. the use of LOC for the detection of exogenous DNA in meat via on-chip PCR in real-time. LOC surfaces were treated with different coatings in order to optimize the DNA extraction directly from meat homogenates (bovine, pork, horse). On the same LOC used for DNA purification, we set up the on-chip PCR with real-time detection. Over 1,000 beef genomes, up to 0.01 horse or pork genomes were successfully detected in binary mixtures of pre-purified DNA and similarly, up to 0.01 % parts of exogenous meat were detected in binary mixtures of meat homogenates. The successful on-chip detection of exogenous DNA is a promising step toward the production of an effective microdevice for rapid, sensitive, and reliable identification of meat adulteration.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Haunshi, S., Basumatary, R., Girish, P. S., Doley, S., Bardoloi, R. K., Kumar, A. (2009). Identification of chicken, duck, pigeon and pig meat by species-specific markers of mitochondrial origin. Meat Science, 83, 454–459.
Hellberg, R. S. R., & Morrissey, M. T. (2011). Advances in DNA-based techniques for the detection of seafood species substitution on the commercial market. Journal of Laboratory Automation, 16, 308–321.
Yin, R. H., Bai, W. L., Wang, J. M., Wu, C. D., Dou, Q. L., Yin, R. L., et al. (2009). Development of an assay for rapid identification of meat from yak and cattle using polymerase chain reaction technique. Meat Science, 83, 38–44.
Rojas, M., González, I., Pavón, M. A., Pegels, N., Hernández, P. E., García, T., et al. (2010). Polymerase chain reaction assay for verifying the labeling of meat and commercial meat products from game birds targeting specific sequences from the mitochondrial D-loop region. Poultry Science, 89, 1021–1032.
Murugaiah, C., Noor, Z. M., Mastakim, M., Bilung, L. M., Selamat, J., Radu, S. (2009). Meat species identification and Halal authentication analysis using mitochondrial DNA. Meat Science, 83, 57–61.
Unajak, S., Meesawat, P., Anyamaneeratch, K., Anuwareepong, D., Srikulnath, K., Choowongkomon, K. (2011). Identification of species (meat and blood samples) using nested-PCR analysis of mitochondrial DNA. African Journal of Biotechnology, 10(29), 5670–5676.
Kesmen, Z., Gulluce, A., Sahin, F., Yetim, H. (2009). Identification of meat species by TaqMan-based real-time PCR assay. Meat Science, 82, 444–449.
Rojas, M., González, I., Pavón, M. Á., Pegels, N., Hernández, P. E., García, T., et al. (2011). Development of a real-time PCR assay to control the illegal trade of meat from protected capercaillie species (Tetrao urogallus). Forensic Science International, 210, 133–138.
Dalton, D. L., & Kotze, A. (2011). DNA barcoding as a tool for species identification in three forensic wildlife cases in South Africa. Forensic Science International, 207, e51–e54.
Ali, M. E., Hashim, U., Mustafa, S., Man, Y. B. C., Yusop, M. H. M., Bari, M. F., et al. (2011). Nanoparticle sensor for label free detection of swine DNA in mixed biological samples. Nanotechnology, 22, 195503.
Zhang, C., & Xing, D. (2010). Single-molecule DNA amplification and analysis using microfluidics. Chemical Reviews, 110, 4910–4947.
Kovarik, M. L., Gach, P. C., Ornoff, D. M., Wang, Y., Balowski, J., Farrag, L., et al. (2012). Micro total analysis systems for cell biology and biochemical assays. Analytical Chemistry, 84(2), 516–540.
Li, Z., Han, D. K., Brokken-Zijp, J. C., de With, G., Thne, P. C. (2006). Surface properties of poly(dimethylsiloxane)-based inorganic/organic hybrid materials. Polymer, 47, 1150–1158.
Crevillén, A. G., Hervás, M., López, M. A., González, M. C., Escarpa, A. (2007). Real sample analysis on microfluidic devices. Talanta, 74, 342–357.
Han, S. W., Joo, S. W., Ha, T. H., Kim, Y., Kim, K. (2000). Adsorption characteristics of anthraquinone-2-carboxylic acid on gold. The Journal of Physical Chemistry. B, 104, 11987–11995.
Schindelin, J., Arganda-Carreras, I., Frise, E., Kaynig, V., Longair, M., Pietzsch, T., et al. (2012). Fiji: an open-source platform for biological-image analysis. Nature Methods, 9, 676–682.
Sharma, S., Johnson, R. W., Desai, T. A. (2004). XPS and AFM analysis of antifouling PEG interfaces for microfabricated silicon biosensors. Biosensors and Bioelectronics, 20, 227–239.
Popat, K. C., Sharma, S., Desai, T. A. (2004). Quantitative XPS analysis of PEG-modified silicon surfaces. The Journal of Physical Chemistry. B, 108, 5185–5188.
Ballin, N. Z., Vogensen, F. K., Karlsson, A. H. (2012). PCR amplification of repetitive sequences as a possible approach in relative species quantification. Meat Science, 90, 438–443.
Potrich, C., Lunelli, L., Pasquardini, L., Sonn, D., Vozzi, D., Dallapiccola, R., et al. (2012). One-shot genetic analysis in monolithic silicon/Pyrex microdevices. Biomedical Microdevices. doi:10.1007/s10544-012-9676-1.
Govindarajan, A. V., Ramachandran, S., Vigil, G. D., Yager, P., Boehringer, K. F. (2012). A low cost point-of-care viscous sample preparation device for molecular diagnosis in the developing world; an example of microfluidic origami. Lab on a Chip, 12, 174–181.
Xu, G., Lee, D. Y. S., Xie, H., Chiew, D., Hsieh, T.-M., Ali, E. M., et al. (2011). A self-contained polymeric cartridge for automated biological sample preparation. Biomicrofluidics, 5, 034107.
Hoffmann, J., Mark, D., Lutz, S., Zengerle, R., von Stetten, F. (2010). Pre-storage of liquid reagents in glass ampoules for DNA extraction on a fully integrated lab-on-a-chip cartridge. Lab on a Chip, 10, 1480–1484.
Nakagawa, T., Tanaka, T., Niwa, D., Osaka, T., Takeyama, H., Matsunaga, T. (2005). Fabrication of amino silane-coated microchip for DNA extraction from whole blood. Journal of Biotechnology, 116, 105–111.
Pasquardini, L., Lunelli, L., Potrich, C., Marocchi, L., Fiorilli, S., Vozzi, D., et al. (2011). Organo-silane coated substrates for DNA purification. Applied Surface Science, 257, 10821–10827.
Emmenegger, C. R., Brynda, E., Riedel, T., Sedlakova, Z., Houska, M., Alles, A. B. (2009). Interaction of blood plasma with antifouling surfaces. Langmuir, 25(11), 6328–6333.
Michel, R., Pasche, S., Textor, M., Castner, D. G. (2005). Influence of PEG architecture on protein adsorption and conformation. Langmuir, 21, 12327–12332.
Pasquardini, L., Potrich, C., Quaglio, M., Lamberti, A., Guastella, S., Lunelli, L., et al. (2011). Solid phase DNA extraction on PDMS and direct amplification. Lab on a Chip, 11, 4029–4035.
Acknowledgments
We acknowledge Olivetti I-Jet for substrate and microchip provision. We are also grateful to Dr. Vanzetti for XPS support.
Author information
Authors and Affiliations
Corresponding author
Electronic Supplementary Material
Below is the link to the electronic supplementary material.
ESM 1
(DOCX 108 kb)
Rights and permissions
About this article
Cite this article
Potrich, C., Santini, G.C., Lunelli, L. et al. The Making of “on-Chip PCR in Real-Time” for Food Quality Control. BioNanoSci. 3, 123–131 (2013). https://doi.org/10.1007/s12668-013-0080-y
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12668-013-0080-y