Development of event-specific PCR detection methods for genetically modified tomato Huafan No. 1.
BACKGROUND
The genetically modified (GM) tomato Huafan No. 1 is a commercial GM event created in China, and the development of specific methods for its identification and quantification is necessary under labeling regulations for genetically modified organisms (GMOs). The event-specific polymerase chain reaction (PCR) method is the most used method for identification of GMOs in routine analysis.


RESULTS
The 3' junction sequence of transgene integration in GM Huafan No. 1 tomato was revealed by thermal asymmetric interlaced PCR (TAIL-PCR) and sequencing analysis. Based on the revealed 3' integration junction sequence, both conventional and real-time PCR assays were developed and validated for GM Huafan No. 1 tomato identification. In the conventional PCR assay, the limit of detection (LOD) was 20 haploid genome copies. In the real-time PCR assay, the LOD and limit of quantification (LOQ) were estimated to be 5 and 10 tomato haploid genome copies, respectively. Furthermore, the developed PCR methods were well validated by in-house validation.


CONCLUSIONS
Our results suggest that the developed event-specific PCR methods can be routinely used for identification and quantification of GM Huafan No. 1 tomato.
National Center for Molecular Characterization of Genetically Modified Organisms, SJTU-Bor Luh Food Safety Center, School of life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
Chao
C
Yang
Dabing
D
Zhang
Litao
L
Yang
D001483:Base Sequence; D002681:China; D018744:DNA, Plant / Q000032:analysis / Q000737:chemistry; D004274:DNA, Recombinant / Q000032:analysis / Q000737:chemistry; D005638:Fruit / Q000235:genetics*; D018551:Solanum lycopersicum / Q000235:genetics*; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results
D018744:DNA, Plant; D004274:DNA, Recombinant