Screening for six genetically modified soybean lines by an event-specific multiplex PCR method: Collaborative trial validation of a novel approach for GMO detection

Grohmann, Lutz; Belter, Anke; Speck, Brigitte; Goerlich, Ottmar; Guertler, Patrick; Angers-Loustau, Alexandre; Patak, Alex

doi:10.1007/s00003-016-1056-y

Cited by 9 publications

(4 citation statements)

References 23 publications

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“…With the rapid development of GM crops, to protect consumer’s right of knowing information about GMO products, the detection methods with simplicity, rapidity, and specificity are still needed to be developed. Transgene crop detection usually adopts molecular detection methods such as cPCR, , qPCR, , multi-qPCR, and dPCR. , The premise of detection is to extract gDNA, while DNA extraction methods are usually purification with phenol/chloroform extraction and the extraction process needs harmful reagents and is time-consuming. Additionally, rapid nucleic acid extraction methods have been reported to simplify extraction process, such as those using paper, alumina membrane, silica, cellulose, and Chelex-100 .…”

Section: Discussionmentioning

confidence: 99%

Rapid Visual LAMP Method for Detection of Genetically Modified Organisms

Xing,

Liang,

Dong

et al. 2023

ACS Omega

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We developed a novel loop-mediated isothermal amplification (LAMP) method using DNA captured on polyacrylamide microparticles (PAMMPs) as templates (PAMMPs@DNA-LAMP) for rapid qualitative detection of genetically modified organisms (GMOs). Here, DNA was extracted by a fast and cost-effective method using PAMMPs. Four LAMP primers were designed for the PAMMPs@DNA-LAMP method to detect the cauliflower mosaic virus 35S (CaMV35S) promotor in GMOs. We thus developed this method for rapid extraction of DNA (5–10 min) and fast amplification of DNA within ∼30 min at a constant temperature of 63 °C. Moreover, the DNA captured by PAMMPs (PAMMPs@DNA) could be effectively detected by both conventional and quantitative PCR (qPCR) and LAMP. The PAMMPs@DNA-LAMP method was validated with high specificity, sensitivity, and performance for practical sample analysis. This assay detected 0.01% target sequences, which had a high specificity like qPCR and better than the conventional PCR (cPCR). Furthermore, PAMMPs@DNA-LAMP was successfully used to extract and detect DNA from food samples of the major crops (soybean, maize, rice, etc.). In summary, a novel PAMMPs@DNA-LAMP assay has been developed, which has higher sensitivity and spends less time than the cPCR detection using the conventional DNA extracted process. This method offers a novel approach for rapid detection of GMOs in the field.

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Section: Discussionmentioning

confidence: 99%

Rapid Visual LAMP Method for Detection of Genetically Modified Organisms

Xing,

Liang,

Dong

et al. 2023

ACS Omega

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“…The multiplex detection methods for universal transgenic genes (P-35S, P-RbcS4, T-nos, T-35S, T-pinII, T-E9, T-AHASL, T-ORF23) of 24 LM soybeans approved in many countries had been developed [23]. These methods can be used as primary screening tools to discriminate suspicious samples whether they contain LMO or not, though further analysis is needed to verify event from LMO containing samples and several multiplex PCR methods of LM soybeans were reported [4,5].…”

Section: Discussionmentioning

confidence: 99%

Development and application of a novel multiplex PCR method for four living modified soybeans

Choi

Seol

et al. 2018

Appl Biol Chem

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Since the early 1990s when the first commercialization of living modified organism (LMO), LMO has been developed to improve nutrient quality and productivity of crops. As the self-sufficiency rate of soybean has gradually decreased in South Korea, most of soybeans have been imported. The cultivation and trade of LM crops are regulated in many countries and authorizations for the use are mandatory in most. In South Korea, the cultivation of LM crop is not allowed and unintentional release of LMO into the natural environment is prohibited. In this study, we developed a novel multiplex PCR method for four LM soybean events (CV127, MON87705, FG72 and MON87701) which were approved recently in South Korea. Multiplex PCR primers were designed for PCR amplification of four LMO event-specific fragments, and we analyzed 41 environmental monitoring samples to confirm the efficiency of this method. These results indicated that the multiplex PCR detection method is sufficient for four LM soybeans found in the natural environment. Based on our finding, we suggest that the new technique may be useful as a lead tool for the development of a detection method for various LMO/GMOs.

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“…In addition, we determined LOD 95% by applying a statistical model for calculating the probability of detection (POD) across laboratories. Since its introduction by Uhlig et al [27], this model has already been used several times to determine the sensitivity of real-time PCR assays [32][33][34][35][36][37]. Qualitative results obtained for the seven dilution steps of the DNA isolate from roe deer meat were used to determine the laboratory standard deviation σ L, and the LOD 95% for the median laboratory, as described previously [27].…”

Section: Lod 95% Derived From the Mixed Model For The Pod Curvementioning

confidence: 99%

Real-Time PCR Assay for the Detection and Quantification of Roe Deer to Detect Food Adulteration—Interlaboratory Validation Involving Laboratories in Austria, Germany, and Switzerland

Druml

Uhlig²,

Simon³

et al. 2021

Foods

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Game meat products are particularly prone to be adulterated by replacing game meat with cheaper meat species. Recently, we have presented a real-time polymerase chain reaction (PCR) assay for the identification and quantification of roe deer in food. Quantification of the roe deer content in % (w/w) was achieved relatively by subjecting the DNA isolates to a reference real-time PCR assay in addition to the real-time PCR assay for roe deer. Aiming at harmonizing analytical methods for food authentication across EU Member States, the real-time PCR assay for roe deer has been tested in an interlaboratory ring trial including 14 laboratories from Austria, Germany, and Switzerland. Participating laboratories obtained aliquots of DNA isolates from a meat mixture containing 24.8% (w/w) roe deer in pork, roe deer meat, and 12 meat samples whose roe deer content was not disclosed. Performance characteristics included amplification efficiency, level of detection (LOD95%), repeatability, reproducibility, and accuracy of quantitative results. With a relative reproducibility standard deviation ranging from 13.35 to 25.08% (after outlier removal) and recoveries ranging from 84.4 to 114.3%, the real-time PCR assay was found to be applicable for the detection and quantification of roe deer in raw meat samples to detect food adulteration.

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Screening for six genetically modified soybean lines by an event-specific multiplex PCR method: Collaborative trial validation of a novel approach for GMO detection

Cited by 9 publications

References 23 publications

Rapid Visual LAMP Method for Detection of Genetically Modified Organisms

Rapid Visual LAMP Method for Detection of Genetically Modified Organisms

Development and application of a novel multiplex PCR method for four living modified soybeans

Real-Time PCR Assay for the Detection and Quantification of Roe Deer to Detect Food Adulteration—Interlaboratory Validation Involving Laboratories in Austria, Germany, and Switzerland

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