Event-specific quantitative polymerase chain reaction methods for detection of double-herbicide-resistant genetically modified corn MON 87419 based on the 3′-junction of the insertion site

2021 ◽  
Author(s):  
Likun Long ◽  
Wei Yan ◽  
Congcong Li ◽  
Liming Dong ◽  
Na Liu ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetically Modified ◽  
Quantitative Polymerase Chain Reaction ◽  
Insertion Site ◽  
Chain Reaction ◽  
Herbicide Resistant ◽  
Qualitative Polymerase Chain Reaction ◽  
Relative Standard ◽  
Junction Site ◽  
Polymerase Chain

ABSTRACT MON 87419 was one of the new transgenic corn events developed in US with the trait of herbicide resistance to both dicamba and glyphosate. To monitor unintended release of genetically modified organism in the future, as well as to meet GM-labeling requirements, it is requisite to develop a reliable method for the detection and quantification of MON 87419, an event-specific primer pair was designed to amplify the 3′-junction site between the endogenous genome sequence and the transferred DNA of GM event MON 87419, amplicons of desired size were produced by qualitative polymerase chain reaction (PCR) assay. For the validation of this quantitative method, the mixed samples containing 10%, 1%, and 0.1% MON 87419 ingredient were quantified. The precisions were expressed as relative standard deviations, deviated by 7.87%, 12.94%, and 19.98%, respectively. These results clearly demonstrate that the PCR methods we developed herein can be used for event-specific quantitative testing of the double-herbicide-resistant corn MON 87419.

scholarly journals Quantitative PCR in diagnosing infectious urogenital pathology

2019 ◽  
pp. 107-111 ◽  
Author(s):  
MR Rakhmatulina ◽  
IS Galkina
Keyword(s):  
Polymerase Chain Reaction ◽  
Inflammatory Diseases ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Sexually Transmitted ◽  
Advantages And Disadvantages ◽  
Qualitative Polymerase Chain Reaction ◽  
Microbiological Methods ◽  
Golden Standard ◽  
Polymerase Chain

This article describes the contemporary methods of diagnosing sexually transmitted infections, their advantages and disadvantages, indications for use. The authors describe application of quantitative polymerase chain reaction in diagnosing inflammatory diseases and dysbiotic conditions in men and women. This method, which is currently the “golden standard” in urogenital pathology diagnostics, has undeniable advantages over microbiological methods and qualitative polymerase chain reaction: the preanalytical stage requirements (preservation of quantitative ratios between microorganisms or nucleic acids of microorganisms) are not as strict, the risk of contamination from outside environment and subsequent corruption of the results is significantly smaller, the conditions for all microorganisms, including those impossible and hard to cultivate, are the same sensitivity and specificity-wise, it is possible to sample materials and evaluate microbiota (ratios of microorganisms and their groups) and also possible to collect samples non-invasively, the speed of testing is high.

Detection of nonauthorized genetically modified organisms using differential quantitative polymerase chain reaction: application to 35S in maize

2008 ◽  
Vol 376 (2) ◽  
pp. 189-199 ◽  
Author(s):  
Katarina Cankar ◽  
Valérie Chauvensy-Ancel ◽  
Marie-Noelle Fortabat ◽  
Kristina Gruden ◽  
André Kobilinsky ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Event-Specific Qualitative and Quantitative Polymerase Chain Reaction Methods for Detection of Genetically Modified Rapeseed Ms8Rf3 Based on the Right Border Junctions

2008 ◽  
Vol 91 (1) ◽  
pp. 143-151 ◽  
Author(s):  
Gang Wu ◽  
Yuhua Wu ◽  
Ling Xiao ◽  
Changming Lu
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetically Modified ◽  
Quantitative Polymerase Chain Reaction ◽  
Artificial Chromosome ◽  
Absolute Quantification ◽  
Detection Methods ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
The Absolute ◽  
The Right

Abstract Ms8Rf3 is a genetically modified rapeseed hybrid which is widely cultivated in Canada and exported to some other countries for production of foodstuffs or fodder. In this study, the genomic sequences flanking the right borders of the integrated transgenic sequences in the Ms8Rf3 genome were characterized and showed high similarities with the bacterial artificial chromosome clone of Chinese cabbage. Event-specific qualitative polymerase chain reaction (PCR) methods were established with the primers and probes targeting the junction regions to produce a 123 base pair (bp) product for the Ms8 event and 92 bp for the Rf3 event. The absolute detection limit of qualitative PCR was 2.5 initial template copies for the Ms8 event and 50 copies for the Rf3 event. Quantitiative detection methods were established, with the absolute quantification limit being approximately 25 initial template copies.

scholarly journals Interlaboratory Transfer of a Real-Time Polymerase Chain Reaction Assay for Quantitative Detection of Genetically Modified Maize Event TC-1507

2006 ◽  
Vol 89 (5) ◽  
pp. 1347-1352 ◽  
Author(s):  
José-Luís La Paz ◽  
Nora Garca-Muniz ◽  
Anna Nadal ◽  
Teresa Esteve ◽  
Pere PuigdomÉnech ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Limit Of Detection ◽  
Genetically Modified ◽  
Insertion Site ◽  
Quantitative Detection ◽  
The European Union ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Maize Chloroplast

Abstract A real-time polymerase chain reaction (QPCR) assay was developed for quantitative detection of a genetically modified (GM) maize event TC-1507 and modification to conventional PCR for qualitative purposes. Sequences 5-flanking TC-1507 full-length insert were characterized and showed multiple rearrangements involving insert and maize chloroplast fragments. The event specificity of the TC-1507 assays was based on the detection of transgene and plant rearranged sequences found to 5 flank the insertion site. They were fully specific and exhibited a limit of detection below 10 target copies, allowing consistent detection of 0.1% GM levels. The QPCR was highly linear and efficient and proved adequate for quantification of GM contents, aiming at the fulfillment of legal requirements established in the European Union (i.e., compulsory labeling of TC-1507 levels >0.9%). It satisfactorily determined TC-1507 contents on different matrixes and was successfully transferred to a different laboratory.

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