Detection of genetic modification in cured tobacco leaf: Proficiency testing using polymerase chain reaction-based methods

被引：0
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作者：
Gadani, Ferruccio [1 ]
Ward, Martin [2 ]
Black, Sue [2 ]
Harris, Neil [3 ]
McDowell, David [3 ]
Bolev, Nick [3 ]
机构：
[1] Philip Morris International, Research and Development, Quai Jeanrenaud 56, CH 2000 Neuchâtel, Switzerland
[2] Advanced Technologies (Cambridge) Ltd., 210 Cambridge Science Park, Cambridge CB4 0WA, United Kingdom
[3] LGC Ltd., Queens Rd, Teddington, Middlesex TW11 0LY, United Kingdom
来源：
Journal of AOAC International | 2006年 / 89卷 / 03期
关键词：
The Cooperation Centre for Scientific Research Relative to Tobacco (CORESTA; Paris; France) Task Force Genetically Modified Tobacco - Detection Methods investigated the performance of qualitative and quantitative methods based on the polymerase chain reaction (PCR) for the detection and quantitation of genetically modified (GM) tobacco. In the 4 successful rounds of proficiency testing; the cauliflower mosaic virus 35S RNA promoter (CaMV 35S) and the Agrobacterium tumefaciens nopaline synthase terminator (NOS) were selected as target sequences. Blind-coded reference materials containing from 0.1 to 5.0% and from 0.15 to 4% GM tobacco were used in 2 rounds of qualitative and quantitative PCR; respectively. Eighteen laboratories from 10 countries participated in this study. Considering all methods and 2 rounds; the different laboratories were able to detect GM tobacco at the 0.1% level in 46 out of 58 tests in qualitative assays. The results of the proficiency test indicate that both end point screening and real-time quantitative methods are suitable for the detection of genetically modified organisms in tobacco leaf samples having a GM content of 0.1% or higher. The CORESTA proficiency study represents a first step towards the interlaboratory evaluation of accuracy and precision of PCR-based GM tobacco detection; which may lead to the harmonization of analytical procedures and to the enhancement of comparability of testing results produced by different laboratories;
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页码：693 / 707
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