中文摘要：聚合酶链反应（PCR）技术已成为检测和识别转基因生物的主要方法。多重PCR技术是PCR技术的进化技术，其允许在同一反应中同时扩增多个目标。本文通过基因和品系特异性分析方法开发、优化、验证了多重PCR技术在转基因油菜GT73、Ms8、Rf3和T45品种的定性检测和识别中的作用。研究发现，多重PCR技术具有较高的特异性、灵敏度、选择性和准确性。每个测试水稻品种基因特异性反应的检测限均为0.01%。作者在2012-2015年分析和检测了428中含有转基因油菜的动物饲料。
外文摘要：Polymerase chain reaction (PCR) has become the main method for detection and identification of genetically modified organisms (CMOs). Multiplex PCR is a variation of this technique allowing simultaneous amplification of several targets in the same reactions. In this study, it was developed, optimized and performed in-house validation of the multiplex PCR method for detection and identification of genetically modified rape lines (GT73, Ms8, Rf3 and T45) in two types of assays: gene- and event-specific. The optimized reactions exhibited high specificity, sensitivity, selectivity and accuracy. The limit of detection (LOD) for the gene-specific reaction was 0.01% for each of the tested lines of rape. For the event-specific reaction, LOD was at the same level for most of the GM rape lines except for T45 (LOD = 0.025%). The optimized and validated assays were employed from 2012 to 2015 in our laboratory to analyse 428 samples of animal feedstuffs containing oilseed rape. GM rape was not detected in the analysed feedstuffs until 2013 (only some traces of GM soyabean were found). Since 2014, the GM rape presence has been confirmed in event-specific reactions in more than one third of analysed samples.
外文关键词：genetically modified rape; multiplex PCR; detection; identification; feedstuffs
作者：Mazur, M.; Sieradzki, Z.; Krol, B.; 等
作者单位：Natl Vet Res Inst Pulawy
期刊名称：JOURNAL OF ANIMAL AND FEED SCIENCES
期刊影响因子：1.024
出版年份：2017
出版刊次：6
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