Development and Validation of A 48-Target Analytical Method for High-throughput Monitoring of Genetically Modified Organisms

作者：Li, Xiaofei; Wu, Yuhua; Li, Jun; Li, Yunjing; Long, Likun; Li, Feiwu^*; Wu, Gang

来源：Scientific Reports, 2015, 5(1): 7616.

DOI：10.1038/srep07616

• REAL-TIME PCR
• POLYMERASE-CHAIN-REACTION
• MULTIPLEX-PCR
• SCREENING-ASSAY
• DIGITAL PCR
• QUANTIFICATION
• MAIZE
• FOOD
• OLIGONUCLEOTIDE
• EVENTS

The rapid increase in the number of genetically modified (GM) varieties has led to a demand for high-throughput methods to detect genetically modified organisms (GMOs). We describe a new dynamic array-based high throughput method to simultaneously detect 48 targets in 48 samples on a Fludigm system. The test targets included species-specific genes, common screening elements, most of the Chinese-approved GM events, and several unapproved events. The 48 TaqMan assays successfully amplified products from both single-event samples and complex samples with a GMO DNA amount of 0.05 ng, and displayed high specificity. To improve the sensitivity of detection, a preamplification step for 48 pooled targets was added to enrich the amount of template before performing dynamic chip assays. This dynamic chip-based method allowed the synchronous high-throughput detection of multiple targets in multiple samples. Thus, it represents an efficient, qualitative method for GMO multi-detection.

• 出版日期2015-1-5
• 单位吉林省农业科学院; 中国农业科学院