Sensitive and Rapid Detection of Genetic Modified Soybean (Roundup Ready) by Loop-Mediated Isothermal Amplification

Liu, Mei; Luo, Yan; Tao, Ran; He, Ru; Jiang, Keyong; Wang, Baojie; Wang, Lei

doi:10.1271/bbb.80723

Cited by 33 publications

(18 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In sensitivity evaluations, LAMP was possible to detect 9 copies, but that derive for PCR was 90 copies. Thus, the LAMP method established in this study was 10 times more sensitive than the conventional PCR, which is in agreement with most previous reports (Liu et al 2009, Yang et al 2009, Fukuma et al 2011. Of the 50 field samples in this study 7 samples were identified as B. bronchiseptica positive by LAMP while conventional PCR presented negative results, making LAMP the obvious choice for diagnosing B. bronchiseptica in clinical practice.…”

Section: Discussionsupporting

confidence: 91%

See 1 more Smart Citation

Rapid and sensitive detection of Bordetella bronchiseptica by loop-mediated isothermal amplification (LAMP)

et al. 2013

View full text Add to dashboard Cite

Bordetella bronchiseptica causes acute and chronic respiratory infections in diverse animal species and occasionally in humans. In this study, we described the establishment of a simple, sensitive and cost-efficient loop-mediated isothermal amplification (LAMP) assay for the detection of B. bronchiseptica. A set of primers towards a 235 bp region within the flagellum gene of B. bronchiseptica was designed with online software.. The specificity of the LAMP assay was examined by using 6 porcine pathogens and 100 nasal swabs collected from healthy pigs and suspect infected pigs. The results indicated that positive reactions were confirmed for all B. bronchiseptica and no cross-reactivity was observed from other non-B. bronchiseptica. In sensitivity evaluations, the technique successfully detected a serial dilutions of extracted B. bronchiseptica DNA with a detection limit of 9 copies, which was 10 times more sensitive than that of PCR. Compared with conventional PCR, the higher sensitivity of LAMP method and no need for the complex instrumentation make this LAMP assay a promising alternative for the diagnosis of B. bronchiseptica in rural areas and developing countries where there lacks of complex laboratory services.

show abstract

Section: Discussionsupporting

confidence: 91%

“…LAMP has successfully been used to diagnose pathogenic infections in humans and animals (Liu et al 2009, Yang et al 2009, Fukuma et al 2011. Compared with other assaysÿthe LAMP method has several advantages.…”

Section: Discussionmentioning

confidence: 99%

Rapid and sensitive detection of Bordetella bronchiseptica by loop-mediated isothermal amplification (LAMP)

et al. 2013

View full text Add to dashboard Cite

show abstract

“…The MON89788 LAMP primers were designed based on event-specific sequence of 3′end of exogenous integration to recognize a target sequence of 259 bp (Genebank accession code. GV597339) [18]. LAMP primers for the soybean endogenous reference gene were targeted at the Lectin sequence (Genebank accession code.…”

Section: Primer Design and Specificity Of Lamp Assaymentioning

confidence: 99%

Rapid Detection of Genetically Modified Ingredients in Soybean Products by Real-Time Loop-Mediated Isothermal Amplification

Di¹,

Shi²,

Shen³

et al. 2014

JFNR

View full text Add to dashboard Cite

A real-time turbidimeter loop-mediated isothermal amplification (LAMP) assay was developed for the rapid, sensitive, and convenient detection of genetically modified (GM) ingredients in soybean products obtained from local supermarkets in China. Specific primers were designed based on the published sequence of lectin, nos terminator and endogenous genes of two GM soybean strains (GTS40-3-2 and MON89788). The total genomic DNA of soybean-made products was extracted by cetyltrimethylammonium bromide (CTAB) method. Then, the genomic DNA was used as template for LAMP assay. The results showed that this real-time turbidimeter LAMP assay was specific and sensitive enough to amplify DNA from soybean products. The limit of detection (LOD) of lectin and MON89788 was 0.01% (weight of GM soybean/weight of non-GM soybean), roughly equivalent to four copies of soybean genomic DNA in DNA mixture within one hour; and the LOD of nos and GTS40-3-2 was 0.1%, roughly equivalent to forty copies of soybean genomic DNA within 1 hour. The sensitivity reaches to 0.01%, which meets the requirements of the European Union (EU). In practical application, the total detectable ratio in soybeanmade products was the same as that of PCR, suggesting that LAMP method was accurate and sensitive.

show abstract

“…Thus, this new method has been widely used in clinical diagnosis [ 18 , 19 , 20 , 21 ], food safety inspection [ 22 , 23 ] and species identification [ 24 , 25 ]. Due to its competitive advantage, the LAMP method was used increasingly for GMO detection [ 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 ]. However, this method has never been used for cry2Ab and cry3A detection.…”

Section: Introductionmentioning

confidence: 99%

Development and Application of Loop-Mediated Isothermal Amplification Assays for Rapid Visual Detection of cry2Ab and cry3A Genes in Genetically-Modified Crops

Wang

Long

et al. 2014

IJMS

View full text Add to dashboard Cite

The cry2Ab and cry3A genes are two of the most important insect-resistant exogenous genes and had been widely used in genetically-modified crops. To develop more effective alternatives for the quick identification of genetically-modified organisms (GMOs) containing these genes, a rapid and visual loop-mediated isothermal amplification (LAMP) method to detect the cry2Ab and cry3A genes is described in this study. The LAMP assay can be finished within 60 min at an isothermal condition of 63 °C. The derived LAMP products can be obtained by a real-time turbidimeter via monitoring the white turbidity or directly observed by the naked eye through adding SYBR Green I dye. The specificity of the LAMP assay was determined by analyzing thirteen insect-resistant genetically-modified (GM) crop events with different Bt genes. Furthermore, the sensitivity of the LAMP assay was evaluated by diluting the template genomic DNA. Results showed that the limit of detection of the established LAMP assays was approximately five copies of haploid genomic DNA, about five-fold greater than that of conventional PCR assays. All of the results indicated that this established rapid and visual LAMP assay was quick, accurate and cost effective, with high specificity and sensitivity. In addition, this method does not need specific expensive instruments or facilities, which can provide a simpler and quicker approach to detecting the cry2Ab and cry3A genes in GM crops, especially for on-site, large-scale test purposes in the field.

show abstract

Sensitive and Rapid Detection of Genetic Modified Soybean (Roundup Ready) by Loop-Mediated Isothermal Amplification

Cited by 33 publications

References 10 publications

Rapid and sensitive detection of Bordetella bronchiseptica by loop-mediated isothermal amplification (LAMP)

Rapid and sensitive detection of Bordetella bronchiseptica by loop-mediated isothermal amplification (LAMP)

Rapid Detection of Genetically Modified Ingredients in Soybean Products by Real-Time Loop-Mediated Isothermal Amplification

Development and Application of Loop-Mediated Isothermal Amplification Assays for Rapid Visual Detection of cry2Ab and cry3A Genes in Genetically-Modified Crops

Contact Info

Product

Resources

About