Development and in‐house validation of the event‐specific qualitative and quantitative PCR detection methods for genetically modified cotton MON15985

Jiang, Lingxi; Yang, Litao; Rao, Jun; Guo, Jinchao; Wang, Shu; Liu, Jia; Lee, Seong‐Hun; Zhang, Dabing

doi:10.1002/jsfa.3829

Cited by 17 publications

(12 citation statements)

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“…These results indicated that the event‐specific quantitative PCR assay for Huafan No. 1 was reliable for quantification of GM tomato samples, in comparison with another paper in our laboratory for GM cotton MON15985 …”

Section: Resultsmentioning

confidence: 79%

Development of event‐specific PCR detection methods for genetically modified tomato Huafan No. 1

2012

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Section: Resultsmentioning

confidence: 79%

Development of event‐specific PCR detection methods for genetically modified tomato Huafan No. 1

2012

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“…[14][15][16] An increasing number of countries and regions have passed strict laws and regulations requiring the labeling of GM food products and detailed safety studies of any transgenic crops before they are approved for commercial production. [17][18][19] For food safety evaluation of GM crops, a major principle and guidance named substantial equivalence was proposed by the Food and Agriculture Organization (FAO) and World Health Organization (WHO) in the early 1990s. [20][21][22] According to the substantial equivalence guideline, various methodologies, including cDNA microarray, miRNA fingerprinting, proteomics, metabolomics and toxicological profiling, need to be applied to investigate the differences between transgenic organisms and their counterpart parental or other wild type lines.…”

Section: Resultsmentioning

confidence: 99%

Digital gene expression analysis of mature seeds of transgenic maize overexpressingAspergillus nigerphyA2and its non-transgenic counterpart

Rao¹,

Yang

Wang³

et al. 2013

GM Crops & Food

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The next generation sequencing technologies have been recently used for transcriptome analysis in many organisms because of the decreased sequencing cost and increased sequence output. In this study, we used digital gene expression (DGE) technique to compare the transcriptomic changes in mature seeds between transgenic maize overexpressing Aspergillus niger phyA2 and its non-transgenic counterpart. Deep sequencing of DGE libraries of the transgenic and its non-transgenic counterpart seeds generated 3,783,500 and 3,790,500 reads of 21-nucleotide, respectively, with frequencies spanning over four orders of magnitude. In transgenic maize, 53.97% of the unambiguous signature tags were mapped to the maize B73 reference genome, and 46.47% of genes were detected by at least two reads; in non-transgenic maize, the corresponding numbers were 51.38% and 47.39%. Compared with non-transgenic counterpart, about 12% of detected genes were differentially expressed in the transcriptome of transgenic maize seeds. Among these differentially expressed genes, there were 23 transcription factors in 14 families and no allergen genes. Pathway enrichment analysis revealed that 21 pathways were significantly affected by the transgenic event, in which the pathway involved in protein processing in endoplasmic reticulum was the most significantly affected. Results from this study indicated that both intended and unintended transcriptomic changes occurred in the transgenic maize, thus emphasizing the importance of transcriptome profiling in risk assessment of transgenic events.

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“…Detection and quantification limits refer to the lowest quantity of the target that can be reliably detected and quantified with a probability of ≥ 95%. The absolute limit is the lowest number of initial template copies that can be detected and quantified . To determine the LOD and LOQ of the established event‐specific RT‐PCR assay, series of DNA dilutions containing an estimated average of 100 000, 10 000, 1000, 100 and 10 copies of the EE‐1 brinjal haploid genome per reaction were tested in triplicate in three parallel reactions (Table ).…”

Section: Resultsmentioning

confidence: 99%

Detection and identification of genetically modified EE‐1 brinjal (Solanum melongena) by single, multiplex and SYBR^® real‐time PCR

2012

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BACKGROUND: Brinjal is an important vegetable crop. Major crop loss of brinjal is due to insect attack. Insect‐resistant EE‐1 brinjal has been developed and is awaiting approval for commercial release. Consumer health concerns and implementation of international labelling legislation demand reliable analytical detection methods for genetically modified (GM) varieties. RESULTS: End‐point and real‐time polymerase chain reaction (PCR) methods were used to detect EE‐1 brinjal. In end‐point PCR, primer pairs specific to 35S CaMV promoter, NOS terminator and nptII gene common to other GM crops were used. Based on the revealed 3′ transgene integration sequence, primers specific for the event EE‐1 brinjal were designed. These primers were used for end‐point single, multiplex and SYBR‐based real‐time PCR. End‐point single PCR showed that the designed primers were highly specific to event EE‐1 with a sensitivity of 20 pg of genomic DNA, corresponding to 20 copies of haploid EE‐1 brinjal genomic DNA. The limits of detection and quantification for SYBR‐based real‐time PCR assay were 10 and 100 copies respectively. CONCLUSION: The prior development of detection methods for this important vegetable crop will facilitate compliance with any forthcoming labelling regulations. Copyright © 2012 Society of Chemical Industry

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Development and in‐house validation of the event‐specific qualitative and quantitative PCR detection methods for genetically modified cotton MON15985

Abstract: This study shows that the developed qualitative and quantitative PCR methods are applicable for the identification and quantification of GM cotton MON15985 and its derivates.

Cited by 17 publications

References 10 publications

Development of event‐specific PCR detection methods for genetically modified tomato Huafan No. 1

Development of event‐specific PCR detection methods for genetically modified tomato Huafan No. 1

Digital gene expression analysis of mature seeds of transgenic maize overexpressingAspergillus nigerphyA2and its non-transgenic counterpart

Detection and identification of genetically modified EE‐1 brinjal (Solanum melongena) by single, multiplex and SYBR^® real‐time PCR

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Development and in‐house validation of the event‐specific qualitative and quantitative PCR detection methods for genetically modified cotton MON15985

Abstract: This study shows that the developed qualitative and quantitative PCR methods are applicable for the identification and quantification of GM cotton MON15985 and its derivates.

Cited by 17 publications

References 10 publications

Development of event‐specific PCR detection methods for genetically modified tomato Huafan No. 1

Development of event‐specific PCR detection methods for genetically modified tomato Huafan No. 1

Digital gene expression analysis of mature seeds of transgenic maize overexpressingAspergillus nigerphyA2and its non-transgenic counterpart

Detection and identification of genetically modified EE‐1 brinjal (Solanum melongena) by single, multiplex and SYBR® real‐time PCR

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Product

Resources

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Detection and identification of genetically modified EE‐1 brinjal (Solanum melongena) by single, multiplex and SYBR^® real‐time PCR