Ultra-Sensitive Detection of Genetically Modified Ingredients in Rice-Derived Products Using Real-Time PCR with Locked Nucleic Acid TaqMan Probe

doi:10.7506/spkx1002-6630-201510042

FOOD SCIENCE

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Ultra-Sensitive Detection of Genetically Modified Ingredients in Rice-Derived Products Using Real-Time PCR with Locked Nucleic Acid TaqMan Probe

PAN Guang1, ZHANG Guiming1, CHEN Zhinan2, CHENG Yinghui1, XIANG Caiyu1, BAO Xianyu2, LING Xingyuan1,*

1. Animal and Plant Inspection and Quarantine Technology Center, Shenzhen Entry-Exit Inspection and Quarantine Bureau,
Shenzhen 518045, China; 2. Shenzhen Academy of Inspection and Quarantine, Shenzhen 518010, China

Online:2015-05-25 Published:2015-05-08
Contact: LING Xingyuan

Abstract

Abstract:

This study aimed to establish a specific and ultra-sensitive detection method for trace genetically modified (GMO)
ingredients in rice-derived products. In this study, the CaMV35S promoter, NOS terminator and Cry1A gene included in the
industrial standard for screening the GM components of rice were selected as the targets. By substituting a few nucleotides
of the TaqMan probe with locked nucleic acid (LNA) nucleotide and comparing the performances of these LNA-TaqMan
probes in real-time PCR (RT-qPCR), a novel real-time PCR method based on LNA-TaqMan probe for the above genes and
gene elements was established with high specificity. Compared to the conventional RT-PCR with TaqMan probe, this RTPCR
with LAN-Taqman probe was much more sensitive, had lower limit of detection (LOD) (0.001% by mass) and 1-3 less
Ct value cycles except for Cry1A. This reported new PCR method can be applied for the detection of trace GM components
in rice-derived products that could not be detected with the conventional RT-PCR probe.

Key words: rice-derived products, locked nucleic acid (LNA), LNA-TaqMan probe, real-time PCR

CLC Number:

S511

PAN Guang1, ZHANG Guiming1, CHEN Zhinan2, CHENG Yinghui1, XIANG Caiyu1, BAO Xianyu2, LING Xingyuan1,*. Ultra-Sensitive Detection of Genetically Modified Ingredients in Rice-Derived Products Using Real-Time PCR with Locked Nucleic Acid TaqMan Probe[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201510042.

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Ultra-Sensitive Detection of Genetically Modified Ingredients in Rice-Derived Products Using Real-Time PCR with Locked Nucleic Acid TaqMan Probe

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