LOOP-MEDIATED ISOTHERMAL AMPLIFICATION BASED BIOSENSOR FOR DETECTION OF GENETICALLY MODIFIED (GMO) SOYBEANS

Sumeyra Vural1, Dogukan Kaygusuz2, Sherjeel Shabih2, Stuart J. Lucas3, A. Ozhan Aytekin1, Meltem Elitas2 1Faculty of Engineering and Natural Science, Yeditepe University, 34755, 2Faculty of Engineering and Natural Science, Sabanci University,l 34956, 3Sabanci Universtiy Nanotechnology Research and Application Center (SUNUM), 34956, Istanbul, Turkey

Aim: Our work presents a cheap, user-friendly, loop-mediated isothermal amplification (LAMP)- based biosensor to detect genetically modified organisms (GMO) in food products. Since more than 82% of the total soy cultivation area is currently planted with GM soybean, here we demonstrate a biosensor that extracts, amplifies and detects transgenes present in GM soybean. Our approach can certainly be modified for GMO detection in other foods. Thanks to its simple operation and low cost, not only mothers at home, but also farmers and workers in the fields and factories can easily use it.

Genomic DNA extraction and Materials and Methods: We fabricated our microfluidic purification is performed using a chip prototype using a 3D printer. Manufacturing and commercial DNA extraction kit for operation of our biosensor is cheap and simple. GMO detection.

Specific amplification is provided by LAMP primer sets including two outer (B3 and F3), loop (FLP and BLP), and inner (FIP and BIP) primer sequences. The LAMP reaction was realised using the WarmStart LAMP kit (New England Biolabs), in a total of 25 µl reaction mixture via incubating at 60 ˚C for 40 minutes.

The temperature control on the device was achieved using a heating coil (SS316), heat sensor (DS18B20 Sensor) and a microprocessor (Atmega).

The LAMP products are made visible to the naked eye by interaction of the amplified oligonucleotides with a DNA specific-dye ( HNB and PVA), which is a hyrogel network system. .

Results: LAMP products with GM: MON810, Bt11 and Bt176 Heating temperature reaches 25˚C to 60˚C The samples heated from 30˚C to 80˚C without GM:RRS and MON89788 in 2 minutes. Sensitivity of the system is 0.49˚C°. the blue color intensity changes.

References: Conclusion and Discussion: 1. James, C. (2014). Global Status of LAMP worked and amplified the GM gene. Commercialized Biotech/GM Crops: 2014 A microprocessor based biosensor generated the physical conditions for the LAMP. (Publication). GMO readout was achieved using a hydrogel network system with HNB-PVA. 2. Walker, G T et al. “Strand Displacement Our method provided cheap and user-friendly method compared to other GMO detection systems Amplification--an Isothermal, in Vitro DNA This system allows easy modifications for detection of other organizations. Amplification Technique.” Nucleic Acids Research 20.7 (1992): 1691–1696. Print. Acknowlegments: 3. Notomi, Tsugunori et al. “Loop-Mediated This project has been suported by Yeditepe Universtiy, Sabanci University and Sabanci University Isothermal Amplification of DNA.” Nucleic Nanotechnology Research and Application Center (SUNUM). Acids Research 28.12 (2000): e63. Print.