Rapid and reliable detection and identification of GM events using multiplex PCR coupled with oligonucleotide microarray.
To devise a rapid and reliable method for the detection and identification of genetically modified (GM) events, we developed a multiplex polymerase chain reaction (PCR) coupled with a DNA microarray system simultaneously aiming at many targets in a single reaction. The system included probes for screening gene, species reference gene, specific gene, construct-specific gene, event-specific gene, and internal and negative control genes. 18S rRNA was combined with species reference genes as internal controls to assess the efficiency of all reactions and to eliminate false negatives. Two sets of the multiplex PCR system were used to amplify four and five targets, respectively. Eight different structure genes could be detected and identified simultaneously for Roundup Ready soybean in a single microarray. The microarray specificity was validated by its ability to discriminate two GM maizes Bt176 and Bt11. The advantages of this method are its high specificity and greatly reduced false-positives and -negatives. The multiplex PCR coupled with microarray technology presented here is a rapid and reliable tool for the simultaneous detection of GM organism ingredients.
Xu
Xiaodan
X
College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
Li
Yingcong
Y
Zhao
Heng
H
Wen
Si-yuan
SY
Wang
Sheng-qi
SQ
Huang
Jian
J
Huang
Kun-lun
KL
Luo
Yun-bo
YB
D029741:Brassica rapa / Q000235:genetics; D018744:DNA, Plant / Q000032:analysis; D018551:Solanum lycopersicum / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012639:Seeds / Q000737:chemistry; D012680:Sensitivity and Specificity; D013025:Glycine max / Q000235:genetics; D003313:Zea mays / Q000235:genetics
D018744:DNA, Plant