A high-throughput liquid bead array-based screening technology for Bt presence in GMO manipulation

Fu, Wei; Wang, Huiyu; Wang, Chenguang; Mei, Lin; Lin, Xiaoyang; Han, Xueqing; Zhu, Shuifang

doi:10.1016/j.bios.2015.10.043

Cited by 20 publications

(9 citation statements)

References 36 publications

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“…In the second category, the expression products of foreign genes were tested by immunological methods or mass spectrometry. 10 reaction and a screen-printed carbon electrode, Ocanã et al 11 have developed a bioanalytical device for detecting 5enolpyruvylshikimate-3-phosphate synthase from Agrobacterium species strain CP4 (CP4-EPSPS) with a limit of detection (LOD) of 0.9 wt %. Although these existing analytical methods applied on GM crops have the merits of high sensitivity and reliability, they are still limited in practice for monitoring GM crops in remote areas.…”

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confidence: 99%

“…have established a duplex droplet digital PCR assay to analyze the endogenous soybean genes and GM-soybean MON 40-3-2 simultaneously, and they achieved the quantitative detection of GM ingredients in complex samples. In the second category, the expression products of foreign genes were tested by immunological methods or mass spectrometry . For example, by combining an immunological reaction and a screen-printed carbon electrode, Ocaña et al have developed a bioanalytical device for detecting 5-enolpyruvylshikimate-3-phosphate synthase from Agrobacterium species strain CP4 (CP4-EPSPS) with a limit of detection (LOD) of 0.9 wt %.…”

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confidence: 99%

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CRISPR-Cas12a-Assisted Multicolor Biosensor for Semiquantitative Point-of-Use Testing of the Nopaline Synthase Terminator in Genetically Modified Crops by Unaided Eyes

et al. 2020

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Existing methods of detecting foreign genes and their expression products from genetically modified organisms (GMOs) suffer from the requirement of professional equipment and skillful operators. The same problem stays for the CRISPR-Cas12a system, although it has been emerging as a powerful tool for nucleic acid detection due to its remarkable sensitivity and specificity. In this report, a portable platform for the visible detection of GMOs based on CRISPR-Cas12a was established, which relies on a color change of gold nanorods (GNRs) caused by the invertase-glucose oxidase cascade reaction and the Fenton reaction for signal readout. A nopaline synthase (NOS) terminator was employed as a model target commonly existing in foreign genes of GMOs. With the help of recombinase-aided amplification, this platform achieved comparable sensitivity of DNA targets (1 aM) with that of a fluorescence reporting assay. As low as 0.1 wt % genetically modified (GM) content in Bt-11 maize was visually observed by unaided eyes, and the semiquantitation of GM ingredients can be obtained within the range of 0.1 to 40 wt % through the absorption measurement of GNRs. Furthermore, five real samples were tested by our method, and the results indicated that the GM ingredient percentages of GMO samples were 2.24 and 24.08 wt %, respectively, while the other three samples were GMO-free. With the advantages of a simple procedure, no need for large or professional instruments, high sensitivity, and selectivity, this platform is expected to provide reasonable technical support for the safe supervision of GMOs.

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confidence: 99%

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confidence: 99%

CRISPR-Cas12a-Assisted Multicolor Biosensor for Semiquantitative Point-of-Use Testing of the Nopaline Synthase Terminator in Genetically Modified Crops by Unaided Eyes

et al. 2020

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“…This suspension array was capable of simultaneously detecting 10 Cry proteins from commercially available GMOs. A novel competitive ELISA using biotinylated antibodies and bead-coupled antigens was established for Cry1Aa-Ab-Ac-Ah, Cry1B, Cry1C, Cry1F, Cry2A, Cry3, and Cry9C in a liquid bead array setup, which has a surprisingly vast range that can be used to test >90% GMOs . Fu et al reported a strategy that could detect several toxins all at once based on the use of commercial antibodies .…”

Section: Fluorescence Immunosensormentioning

confidence: 99%

Advances in the Immunoassays for Detection of Bacillus thuringiensis Crystalline Toxins

Faheem

Qin

Nan

et al. 2021

J. Agric. Food Chem.

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Insect-resistant genetically modified organisms have been globally commercialized for the last 2 decades. Among them, transgenic crops based on Bacillus thuringiensis crystalline (Cry) toxins are extensively used for commercial agricultural applications. However, less emphasis is laid on quantifying Cry toxins because there might be unforeseen health and environmental concerns. Immunoassays, being the preferred method for detection of Cry toxins, are reviewed in this study. Owing to limitations of traditional colorimetric enzyme-linked immunosorbent assay, the trend of detection strategies shifts to modified immunoassays based on nanomaterials, which provide ultrasensitive detection capacity. This review assessed and compared the properties of the recent advances in immunoassays, including colorimetric, fluorescence, chemiluminescence, surface-enhanced Raman scattering, surface plasmon resonance, and electrochemical approaches. Thus, the ultimate aim of this study is to identify research gaps and infer future prospects of current approaches for the development of novel immunosensors to monitor Cry toxins in food and the environment.

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“…One alternative strategy is to use certain affinity ligands comprising aptamers , or antibodies − that specifically interact with GM event biomarkers for GMO assay. Among them, the protein-based immunoassays, such as ELISA , and immunochromatographic test strip, display concrete advantages in simple manipulation, no need for sophisticated instruments, and low cost. Especially, the immunochromatographic test strip has the superiority of fitting for field tests.…”

Section: Introductionmentioning

confidence: 99%

Sensitive and Facile Electrochemiluminescent Immunoassay for Detecting Genetically Modified Rapeseed Based on Novel Carbon Nanoparticles

Gao

Wen

et al. 2018

J. Agric. Food Chem.

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A highly sensitive electrochemiluminescent (ECL) immunoassay targeting PAT/ bar protein was facilely developed for genetically modified (GM) rapeseed detection using carbon nanoparticles (CNPs) originally prepared from printer toner. In this work, CNPs linked with antibody for PAT/ bar protein were used to modify a working electrode. After an immunoreaction between the PAT/ bar protein and its antibody, the immunocomplex formed on the electrode receptor region resulted in an inhibition of electron transfer between the electrode surface and the ECL substance, thus led to a decrease of ECL response. Under the optimal conditions, the ECL responses linearly decreased as the increase of the PAT/ bar protein concentration and the GM rapeseed RF3 content in the ranges of 0.10-10 ng/mL and 0.050-1.0%, with the limits of detection of 0.050 ng/mL and 0.020% (S/N = 3). These results open a facile, sensitive, and rapid approach for the safety control of agricultural GM rape.

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A high-throughput liquid bead array-based screening technology for Bt presence in GMO manipulation

Cited by 20 publications

References 36 publications

CRISPR-Cas12a-Assisted Multicolor Biosensor for Semiquantitative Point-of-Use Testing of the Nopaline Synthase Terminator in Genetically Modified Crops by Unaided Eyes

CRISPR-Cas12a-Assisted Multicolor Biosensor for Semiquantitative Point-of-Use Testing of the Nopaline Synthase Terminator in Genetically Modified Crops by Unaided Eyes

Advances in the Immunoassays for Detection of Bacillus thuringiensis Crystalline Toxins

Sensitive and Facile Electrochemiluminescent Immunoassay for Detecting Genetically Modified Rapeseed Based on Novel Carbon Nanoparticles

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