Surface Plasmon Resonance Detection of Transgenic Cry1Ac Cotton (Gossypium spp.)

Abstract

The detection and identification of genetically modified (GM) plants are challenging issues that have arisen from the potential negative impacts of extensive cultivation of transgenic plants. The screening process is a long-term focus and needs specific detection strategies. Surface plasmon resonance (SPR) has been used to detect a variety of biomolecules including proteins and nucleic acids due to its ability to monitor specific intermolecular interactions. In the present study, two high-throughput, label-free, and specific methods based on SPR technology were developed to detect transgenic Cry1Ac cotton ( Gossypium spp.) by separately targeting protein and DNA. In the protein-based detection system, monoclonal anti-Cry1Ac antibodies were immobilized on the surface of a CM5 sensor chip. Conventional cotton samples were used to define the detection threshold. Transgenic cotton was easily identified within 5 min per sample. For the DNA-based model, a 25-mer biotinylated oligonucleotide probe was immobilized on an SA sensor chip. PCR products of Cry1Ac (230 bp) were used to investigate the reaction conditions. The sensitivity of the constructed sensor chip was identified at concentrations as low as 0.1 nM based on its complementary base pairing.

Advances in the Immunoassays for Detection of Bacillus thuringiensis Crystalline Toxins

“…These SERS-barcoded nanoparticles are a potent tool for multiplex immunosensing in complex environments having traces of analytes. Later, a DNA-based SPR sensor identified Cry1Ac toxin up to 0.1 nM depending upon complementary base pairing with a polymerase chain reaction (PCR)-amplified sequence, whereas anti-Cry1Ac mAb on a CM5 sensor chip provides convenient and rapid genetically modified (GM) cotton detection in 5 min …”

“…Later, a DNAbased SPR sensor identified Cry1Ac toxin up to 0.1 nM depending upon complementary base pairing with a polymer-ase chain reaction (PCR)-amplified sequence, whereas anti-Cry1Ac mAb on a CM5 sensor chip provides convenient and rapid genetically modified (GM) cotton detection in 5 min. 69 The SPR phenomenon refers to the insignificant electromagnetic field generated by electrons between the positive and negative permittivity material after excitation by a beam of incident light at a particular wavelength and angle. 70 The concept of the SPR sensor was investigated quite intensively in recent years.…”

Transgenic Plant Detection Using an AuNPs Based SPR Biosensor

“…The results revealed a detection limit of 1 nM of the complementary fragments using the SPR system with tbiotinylated oligonucleotide probes immobilized on the sensor chip. In another study, an SPR method was developed to detect transgenic Cry1Ac cotton [30]. An SA-coated sensor chip was used to immobilize a 25-mer biotinylated probe, and the detection conditions were determined using Cry1Ac (230 bp) PCR products.…”

Phage-Mediated Immuno-PCR for Ultrasensitive Detection of Cry1Ac Protein Based on Nanobody

“…Currently, the use of transgenic plants (including tobacco, cotton, rice, potato, and maize plants) producing Cry proteins has rapidly increased. For example, as of 2015, 81% of maize and 84% of cotton planted in the United States expressed one or more insecticidal traits for protection against a diversity of insect pests. , However, the widespread use of Cry proteins in transgenic plants for insect control has raised concerns about the problem of gene flow, insect resistance, and allergenicity caused by Cry proteins. − As such, a rapid, sensitive, and selective analytical method for introduced genes or their expressed proteins is important for environmental and food safety. , …”