Detection of genetically modified maize MON810 and NK603 by multiplex and real-time polymerase chain reaction methods.
In this study, the event-specific primers for insecticide-resistant maize, MON810, and herbicide-tolerance maize, NK603, have been designed. Simplex PCR and multiplex PCR detection method have been developed. The detection limit of the multiplex PCR is 0.5% for MON810 and NK603 in 50 ng of the template for one reaction. Quantitative methods based on real-time quantitative PCR were developed for MON810 and NK603. Plasmid pMulM2 as reference molecules for the detection of MON810 and NK603 was constructed. Quantification range was from 0.5 to 100% in 100 ng of the DNA template for one reaction. The precision of real-time Q-PCR detection methods, expressed as coefficient of variation for MON810 and NK603 varied from 1.97 to 8.01% and from 3.45 to 10.94%, respectively. The range agreed with European interlaboratories test results (25%). According to the results, the methods for quantitative detection of genetically modified maize were acceptable.
Huang
Hsin-Ying
HY
Institute of Microbiology and Biochemistry, National Taiwan University, Taipei, Taiwan 106.
Pan
Tzu-Ming
TM
D018744:DNA, Plant / Q000032:analysis*; D030821:Plants, Genetically Modified / Q000145:classification / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012015:Reference Standards; D012680:Sensitivity and Specificity; D003313:Zea mays / Q000145:classification / Q000235:genetics*
D018744:DNA, Plant