Multiplex real-time PCR Method for the detection and quantification of DNA from the four transgenic soy traits DP-356043-5, DP-305423-1, MON 87701, and BPS-CV127-9 and lectin

被引:6
作者
Koeppel, Rene [1 ]
Bucher, Thomas [1 ]
Meuwly, Alain [2 ]
Moor, Dominik [2 ]
机构
[1] Official Food Control Author Canton Zurich, Zurich, Switzerland
[2] Fed Off Publ Hlth, Bern, Switzerland
关键词
Multiplex; Quantitative real-time PCR; DP-356043-5; DP-305423-1; MON; 87701; BPS-CV127-9;
D O I
10.1007/s00217-014-2229-5
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
According to the release of authorization, several new transgenic soy crops are expected to be present in the harvest of soy beans. In contrast to earlier times, methods for the detection of these transgenic crops are available already in advance. However, these methods are mainly still inefficient single real-time PCR methods. To increase the efficiency of product control, a multiplex quantitative real-time PCR system was developed and characterized for the four new transgenic soy traits DP-356043-5, DP-305423-1, MON 87701 and BPS-CV127-9. It showed amplification efficiency, correlation and sensitivity similar to the single PCR systems applied therein. To evaluate the robustness, an appropriate testing scheme was developed and applied for the first time on this multiplex real-time PCR system. It showed the robust amplification of all analytes also in case where conditions were varied. This system allows relative multiplex quantification and/or delta-delta Ct method quantification and proofed the applicability in routine.
引用
收藏
页码:347 / 355
页数:9
相关论文
共 17 条
[1]  
[Anonymous], DEF MIN PERF REQ AN
[2]  
[Anonymous], 215692005 ISO
[3]  
[Anonymous], 2005, 215702005 ISO
[4]  
[Anonymous], EV SPEC METH QUANT S
[5]  
[Anonymous], EV SPEC METH QUANT S
[6]  
Arumuganathan K., 1991, Plant Mol Biol Rep, V9, P208, DOI [10.1007/BF02672069, DOI 10.1007/BF02672069]
[7]   The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments [J].
Bustin, Stephen A. ;
Benes, Vladimir ;
Garson, Jeremy A. ;
Hellemans, Jan ;
Huggett, Jim ;
Kubista, Mikael ;
Mueller, Reinhold ;
Nolan, Tania ;
Pfaffl, Michael W. ;
Shipley, Gregory L. ;
Vandesompele, Jo ;
Wittwer, Carl T. .
CLINICAL CHEMISTRY, 2009, 55 (04) :611-622
[8]  
European Network of GMO Laboratories, 2011, VER REAL TIM PCR MET
[9]  
European Network of GMO Laboratories (ENGL), 2008, DEF MIN PERF REQ AN
[10]  
International Organization for Standardization, 2005, ISO IEC 17025GENERAL