Droplet digital PCR versus multiplex real-time PCR method for the detection and quantification of DNA from the four transgenic soy traits MON87769, MON87708, MON87705 and FG72, and lectin

被引:31
作者
Koeppel, Rene [1 ]
Bucher, Thomas [1 ]
Frei, Anna [1 ]
Waiblinger, Hans-Ulrich [2 ]
机构
[1] Official Food Control Author Canton Zurich, Zurich, Switzerland
[2] State Inst Chem & Veterinarian Anal, Freiburg, Germany
关键词
Digital multiplex PCR; Quantitative real-time PCR; MON87769; MON87708; MON87705; FG72; GENETICALLY-MODIFIED MAIZE;
D O I
10.1007/s00217-015-2481-3
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
New genetically modified (gm) soy crops are going to be released for human consumption. Therefore, analytical skills have to be developed towards an efficient detection and determination of GMO content in feed and food. Existing approaches to screen gm plants do not detect new gm soy traits in many cases. Therefore, a multiplex quantitative real-time PCR system was developed and characterized for the four new transgenic soy traits MON87769, MON87708, MON87705 and FG72 to avoid time- and cost-consuming application of single event detection. It showed amplification efficiency, correlation and limit of quantification similar to the single PCR systems applied. The droplet digital PCR showed increased specificity. In parallel, we developed four duplex droplet digital PCR systems and compared the results from both methods. This showed that both approaches are fit for routine diagnostics. Real-time PCR may be more suited for screening as it is very cost-efficient. Digital PCR may be more suitable for quantitative analysis as it exhibited a measurement uncertainty of only 17 % or below for a single reaction.
引用
收藏
页码:521 / 527
页数:7
相关论文
共 17 条
[1]  
[Anonymous], 2012, EV SPEC METH QUANT S
[2]  
[Anonymous], 2013, EV SPEC METH QUANT S
[3]   The applicability of digital PCR for the assessment of detection limits in GMO analysis [J].
Burns, M. J. ;
Burrell, A. M. ;
Foy, C. A. .
EUROPEAN FOOD RESEARCH AND TECHNOLOGY, 2010, 231 (03) :353-362
[4]   Absolute quantification of genetically modified MON810 maize (Zea mays L.) by digital polymerase chain reaction [J].
Corbisier, Philippe ;
Bhat, Somanath ;
Partis, Lina ;
Xie, Vicki Rui Dan ;
Emslie, Kerry R. .
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2010, 396 (06) :2143-2150
[5]  
European Network of GMO Laboratories, 2011, VER REAL TIM PCR MET
[6]   Interlaboratory transfer of a PCR multiplex method for simultaneous detection of four genetically modified maize lines:: Bt11, MON810, T25, and GA21 [J].
Hernández, M ;
Rodríguez-Lázaro, D ;
Zhang, D ;
Esteve, T ;
Pla, M ;
Prat, S .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 2005, 53 (09) :3333-3337
[7]   Absolute quantification by droplet digital PCR versus analog real-time PCR [J].
Hindson, Christopher M. ;
Chevillet, John R. ;
Briggs, Hilary A. ;
Gallichotte, Emily N. ;
Ruf, Ingrid K. ;
Hindson, Benjamin J. ;
Vessella, Robert L. ;
Tewari, Muneesh .
NATURE METHODS, 2013, 10 (10) :1003-+
[8]   Reliable detection and identification of genetically modified maize, soybean, and canola by multiplex PCR analysis [J].
James, D ;
Schmidt, AM ;
Wall, E ;
Green, M ;
Masri, S .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 2003, 51 (20) :5829-5834
[9]   Multiplex real-time PCR for the detection and quantification of DNA from four transgenic soy Mon89788, A5547-127, Roundup Ready, A2704-12 and lectin [J].
Koeppel, Ren ;
van Velsen, Franziska ;
Felderer, Nora ;
Bucher, Thomas .
EUROPEAN FOOD RESEARCH AND TECHNOLOGY, 2012, 235 (01) :23-28
[10]   Rapid establishment of droplet digital PCR for quantitative GMO analysis [J].
Koeppel, Rene ;
Bucher, Thomas .
EUROPEAN FOOD RESEARCH AND TECHNOLOGY, 2015, 241 (03) :427-439