Rapid detection of P–35S and T-nos in genetically modified organisms by recombinase polymerase amplification combined with a lateral flow strip

Liu, Hua; Wang, Jinbin; Li, Peng; Bai, Liang; Jia, Junjie; Pan, Aihu; Long, XuanQi; Cui, Wei-Dong; Tang, Xueming

doi:10.1016/j.foodcont.2019.106775

Cited by 31 publications

(10 citation statements)

References 34 publications

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“…Portable suitcase / 0.1% [32] DNAzyme-enhanced LFB 120 min 0.1% [33] RPA-based LFD / 50 and 100 copies [34] CPA-based anti-contamination test paper / / [35] More efficient high-throughput and convenient detection methods should be established in the future to promote and optimize large-scale screening in the field. Furthermore, the rapid pre-treatment of deep-processed foods containing GM ingredients in the market warrants investigation, while effective contamination prevention, visualization, and quantitative detection in the field are also significant and promising directions for future development.…”

Section: Discussionmentioning

confidence: 99%

“…For the identification of P-35S and T-nos in GMOs, Liu et al established an RPA-based and lateral flow test paper (LFD)-based platform. Following the labeling of forward and reverse primers with various fluorescent groups at the 5 end, samples were quickly assessed for the presence of P-35S and T-nos genes of GMOs using the RPA-LFD technique at room temperature and, in the 9 GMO samples tested, the detection thresholds of RPA-LFD were found to be 50 and 100 copies [34]. Huang et al created a cross-primed isothermal amplification assay (CPA) for the rapid detection of the 35S promoter of the plant pathogen cauliflower mosaic virus (CaMV 35S) in the field, which they combined with a biosensor made of anti-contamination test paper.…”

Section: On-site Detection Biosensor Based On Functional Nucleic Acidmentioning

confidence: 99%

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Advanced Technologies in On-Site Detection of Genetically Modified Products

Lin

et al. 2022

Agriculture

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Transgenic technology is significantly impacting life today. However, with the advancement of genetically modified technologies and the success of genetically modified product commercialization, new challenges have arisen for associated detecting technologies. The need for fast, precise, and portable systems for the on-site detection of genetically modified products has increased dramatically in recent years. This perspective examined the currently available technological support for portable immune biosensing, discussed a portable detection device for ultrafast PCR, and an on-site detection biosensor based on functional nucleic acid and superior detection devices in the field. Moreover, the on-site sequencing of genetically modified organisms was mentioned briefly. Lastly, the future outlook of genetically modified products detection was concluded and discussed in order to provide a comprehensive reference for future research and development in related fields.

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Section: Discussionmentioning

confidence: 99%

Section: On-site Detection Biosensor Based On Functional Nucleic Acidmentioning

confidence: 99%

Advanced Technologies in On-Site Detection of Genetically Modified Products

Lin

et al. 2022

Agriculture

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“…Meanwhile, RPA shows significant advantages, such as a short incubation time, lower incubation temperatures, and a high tolerance to sample impurities. It is possible to perform multiple RPA amplifications in one reaction system, and this has been successfully reported, for example, in a real-time fluorescent duplex RPA assay for the detection of Campylobacter coli and jejuni from eggs and chicken products [37], a duplex SRES-based lateral flow assay for the detection of Listeria monocytogenes and Salmonella Enteritidis [1], and a duplex lateral flow assay for the detection of P-35S and T-nos in genetically modified organisms [38]. However, there have been few reports on the LFD-RPA assay for the simultaneous detection of three or more foodborne pathogens [39].…”

Section: Discussionmentioning

confidence: 99%

Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood

Chen

et al. 2020

Foods

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Foodborne pathogens can cause foodborne illness. In reality, one food sample may carry more than one pathogen. A rapid, sensitive, and multiple target method for bacteria detection is crucial in food safety. For the simultaneous detection of Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella Enteritidis, multi-objective recombinase polymerase amplification (RPA) combined with a lateral flow dipstick (LFD) was developed in this study. The whole process, including amplification and reading, can be completed in 15 min at 37 °C. The detection limits were 2.6 × 101 CFU/mL for Staphylococcus aureus, 7.6 × 101 CFU/mL for Vibrio parahaemolyticus, and 1.29 × 101 CFU/mL for Salmonella Enteritidis. Moreover, colored signal intensities on test lines were measured by a test strip reader to achieve quantitative detection for Staphylococcus aureus (R2 = 0.9903), Vibrio parahaemolyticus (R2 = 0.9928), and Salmonella Enteritidis (R2 = 0.9945). In addition, the method demonstrated good recoveries (92.00%–107.95%) in the testing of spiked food samples. Therefore, the multiplex LFD-RPA assay is a feasible method for the rapid, sensitive, and quantitative detection of bacterial pathogens in seafood.

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“…Among them, RPA has been widely used for nucleic acid detection because of its high efficiency and easily achievable reaction temperature (37 -42 ℃) [15]. Recently, strategies such as RPA-AGE (RPA combined with agarose gel electrophoresis) [16], RT-RPA (real-time fluorescent RPA) [15,17], RPA-SYBR Green I (RPA combined with DNA fluorescent dye) [18,19], RPA-LFD (RPA combined with lateral flow immunochromatography) [20,21] and RPA-Cas12a-FS (RPA combined with Cas12a cutting) [22] have been established and implemented in the field of rapid detection. Among these methods, the test strip method is the most suitable for on-site testing for its full independence of testing equipment.…”

Section: Introductionmentioning

confidence: 99%

A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4- EPSPS and Cry1Ab/ Ac genes in genetically modified crops

Wang¹,

Wang²,

Hu³

et al. 2024

Food Science and Human Wellness

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Traditional transgenic detection methods require high test conditions and struggle to be both sensitive and efficient. In this study, a one-tube dual RPA reaction system for CP4-EPSPS and Cry1Ab/Ac was proposed and combined with a lateral flow immunochromatographic assay, named "Dual-RPA-LFD", to visualize the dual detection of GM crops. In which, the herbicide tolerance gene CP4-EPSPS and the insect resistance gene Cry1Ab/Ac were selected as targets taking into account the current status of the most widespread application of insect resistance and herbicide tolerance traits and their stacked traits. Gradient diluted plasmids, transgenic standards, and actual samples were used as templates to conduct sensitivity, specificity, and practicality assays, respectively. The constructed method achieved the visual detection of plasmid at levels as low as 100 copies, demonstrating its high sensitivity. In addition, good applicability to transgenic samples was observed, with no cross-interference between two test lines and no influence from other genes. In conclusion, this strategy achieved the expected purpose of simultaneous detection of the two popular targets in GM crops within 20 minutes at 37 ℃ in a rapid, equipment-free field manner, providing a new alternative for rapid screening for transgenic assays in the field.

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Rapid detection of P–35S and T-nos in genetically modified organisms by recombinase polymerase amplification combined with a lateral flow strip

Cited by 31 publications

References 34 publications

Advanced Technologies in On-Site Detection of Genetically Modified Products

Advanced Technologies in On-Site Detection of Genetically Modified Products

Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood

A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4- EPSPS and Cry1Ab/ Ac genes in genetically modified crops

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