摘要
标准物质是转基因检测结果可靠性、可比性以及准确性的重要参比,不可缺少。本文研究了植物转基因成分检测中基体标准物质的长期保存对PCR检出及低含量标准物质量值稳定性的影响。应用普通定性PCR、实时荧光PCR和微滴式数字PCR 3种方法,对不同含量转基因标准物质和转基因检测样品进行了复现性和量值稳定检测。结果表明,-70~-75℃保存条件下,9年保存期的转基因含量为0.1%的单一作物转化体标准物质中依然能得到清晰的转化体目标片段扩增,且Ct值在36左右。数字PCR定值结果表明,9年保存期的转基因基体标准物质的量值保持稳定。11年保存期的大豆、玉米、水稻的混合基体样品中各转基因元件均能稳定检出。研究结果表明转基因基体标准物质的长期低温保存不会影响其各转基因元件的检出和作为转基因核酸检测阳性对照的应用。
Reference material is indispensable for the reliability, comparability, and accuracy of transgenic detection results. In this paper, the effects of long-term preservation of matrix reference materials on PCR detection and the stability of low content reference materials were studied. The reproducibility and quantity stability of transgenic reference materials and transgenic samples with different contents were detected by three methods(common qualitative PCR, real-time fluorescent PCR, and droplet digital PCR). The result showed that the transgenic reference material with 0.1% GM content could still be clearly amplified after 9 years of storage, and the Ct value was about 36. The digital PCR revealed that the quantity of GM matrix reference materials remained stable for long storage time. There was no effect on the detection of transgenic elements in sample mixed with soybean,corn and rice powder which stored at-70 to-75℃ f or 11 years. In summary, the long-term storage of GM matrix reference materials at low temperature would not affect the detection of each transgenic component and its application as a positive control for transgenic nucleic acid detection.
作者
王渭霞
赖凤香
胡海燕
何佳春
魏琪
万品俊
傅强
WANG Wei-Xia;LAI Feng-Xiang;HU Hai-Yan;HE Jia-Chun;WEI Qi;WAN Pin-Jun;FU Qiang(China National Rice Research Institute,Hangzhou 310006,Zhejiang,China)
出处
《作物学报》
CAS
CSCD
北大核心
2022年第1期238-248,共11页
Acta Agronomica Sinica
基金
中国农业科学院科技创新工程“水稻病虫草害防控技术科研团队”(CAAS-ASTIP-2016-CNRRI)
中央级公益性科研院所基本科研业务费专项资金(2017RG005)资助。
关键词
转基因生物
基体标准物质
转基因植物检测
标准物质保存
genetically modified organism(GMO)
matrix reference material
detection of transgenic plants
reference material storage
