Interlaboratory Study of DNA Extraction from Multiple Ground Samples, Multiplex Real-Time PCR, and Multiplex Qualitative PCR for Individual Kernel Detection System of Genetically Modified Maize

2011 ◽  
Vol 94 (5) ◽  
pp. 1540-1547 ◽  
Author(s):  
Hiroshi Akiyama ◽  
Kozue Sakata ◽  
Daiki Makiyma ◽  
Kosuke Nakamura ◽  
Reiko Teshima ◽  
...  
2011 ◽  
Vol 94 (5) ◽  
pp. 1540-1547
Author(s):  
Hiroshi Akiyama ◽  
Kozue Sakata ◽  
Daiki Makiyma ◽  
Kosuke Nakamura ◽  
Reiko Teshima ◽  
...  

Abstract In many countries, the labeling of grains, feed, and foodstuff is mandatory if the genetically modified (GM) organism content exceeds a certain level of approved GM varieties. We previously developed an individual kernel detection system consisting of grinding individual kernels, DNA extraction from the individually ground kernels, GM detection using multiplex real-time PCR, and GM event detection using multiplex qualitative PCR to analyze the precise commingling level and varieties of GM maize in real sample grains. We performed the interlaboratory study of the DNA extraction with multiple ground samples, multiplex real-time PCR detection, and multiplex qualitative PCR detection to evaluate its applicability, practicality, and ruggedness for the individual kernel detection system of GM maize. DNA extraction with multiple ground samples, multiplex real-time PCR, and multiplex qualitative PCR were evaluated by five laboratories in Japan, and all results from these laboratories were consistent with the expected results in terms of the commingling level and event analysis. Thus, the DNA extraction with multiple ground samples, multiplex real-time PCR, and multiplex qualitative PCR for the individual kernel detection system is applicable and practicable in a laboratory to regulate the commingling level of GM maize grain for GM samples, including stacked GM maize.


2007 ◽  
Vol 73 (24) ◽  
pp. 8012-8017 ◽  
Author(s):  
S. Wiedemann ◽  
P. Gürtler ◽  
C. Albrecht

ABSTRACT Rumen-cannulated cows (n = 4) were fed successively silage made from either conventional or genetically modified (GM) maize. Results revealed no effects of GM maize on the dynamics of six ruminal bacterial strains (investigated by real-time PCR) compared to the conventional maize silage.


2010 ◽  
Vol 93 (2) ◽  
pp. 734-749 ◽  
Author(s):  
Takashi Kodama ◽  
Yasunori Kurosawa ◽  
Kazumi Kitta ◽  
Shigehiro Naito

Abstract The Horwitz curve estimates interlaboratory precision as a function only of concentration, and is frequently used as a method performance criterion in food analysis with chemical methods. The quantitative biochemical methods based on real-time PCR require an analogous criterion to progressively promote method validation. We analyzed the tendency of precision using a simplex real-time PCR technique in 53 collaborative studies of seven genetically modified (GM) crops. Reproducibility standard deviation (SR) and repeatability standard deviation (Sr) of the genetically modified organism (GMO) amount () was more or less independent of GM crops (i.e., maize, soybean, cotton, oilseed rape, potato, sugar beet, and rice) and evaluation procedure steps. Some studies evaluated whole steps consisting of DNA extraction and PCR quantitation, whereas others focused only on the PCR quantitation step by using DNA extraction solutions. Therefore, SR and Sr for GMO amount () are functions only of concentration similar to the Horwitz curve. We proposed SR 0.1971C0.8685 and Sr 0.1478C0.8424, where C is the GMO amount (). We also proposed a method performance index in GMO quantitative methods that is analogous to the Horwitz Ratio.


2010 ◽  
Vol 3 (4) ◽  
pp. 304-312 ◽  
Author(s):  
Silvia Folloni ◽  
Gianni Bellocchi ◽  
Adelina Prospero ◽  
Maddalena Querci ◽  
William Moens ◽  
...  

2020 ◽  
Vol 9 (2) ◽  
pp. 1
Author(s):  
Eleni Spanea ◽  
Theofania Tsironi ◽  
Efstathia Tsakali ◽  
Anthimia Batrinou ◽  
Valentina Stefanou ◽  
...  

The objective of the study was to determine qualitatively by validated Real Time PCR method the occurrence of genetically modified maize and soybean in commercial food products from the Greek market. 70 independent samples were collected, including products from different categories (i.e. cereal based, biscuits and snacks) which declared either corn or soybean on the labelling. The result of the study indicated that 37.1% of maize and soy products (n=70) displayed in the Greek market have detectable levels of genetically modified maize or soy. These products were identified by specific primers and included common GMΟ detection primers for 35S and NOS terminator. Adequate repeatability and reproducibility was demonstrated for the applied Real Time PCR method, as evaluated by intra- and inter-laboratory tests.


2013 ◽  
Vol 7 (1) ◽  
pp. 224-233 ◽  
Author(s):  
Maher Chaouachi ◽  
Mohamed Salem Zellama ◽  
Nesrine Nabi ◽  
Ahmed Ben Hafsa ◽  
Khaled Saïd

Food Control ◽  
2010 ◽  
Vol 21 (11) ◽  
pp. 1536-1544 ◽  
Author(s):  
Jasbeer Kaur ◽  
Son Radu ◽  
Farinazleen Mohamad Ghazali ◽  
Cheah Yoke Kqueen

2009 ◽  
Vol 50 (3) ◽  
pp. 117-125 ◽  
Author(s):  
Taichi OGUCHI ◽  
Mari ONISHI ◽  
Yasutaka MINEGISHI ◽  
Yasunori KUROSAWA ◽  
Masaki KASAHARA ◽  
...  

2012 ◽  
Vol 37 (11) ◽  
pp. 2117-2121
Author(s):  
Lei YUAN ◽  
Hong-Wei SUN ◽  
Fan LI ◽  
Ning LI ◽  
Lei ZHAO ◽  
...  

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