title: Detection of genetic modification in cured tobacco leaf: proficiency testing using polymerase chain reaction-based methods.
abstract: 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.
authors.affiliation: Philip Morris International, Research and Development, Quai Jeanrenaud 56, CH 2000 Neuchâtel, Switzerland. ferruccio.gadani@pmintl.com
authors.forename: Ferruccio
authors.initials: F
authors.lastname: Gadani
authors.forename: Martin
authors.initials: M
authors.lastname: Ward
authors.forename: Sue
authors.initials: S
authors.lastname: Black
authors.forename: Neil
authors.initials: N
authors.lastname: Harris
authors.forename: David
authors.initials: D
authors.lastname: McDowell
authors.forename: Nick
authors.initials: N
authors.lastname: Boley
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D000594:Amino Acid Oxidoreductases / Q000235:genetics; D017796:Caulimovirus / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D005188:False Negative Reactions; D018515:Plant Leaves / Q000378:metabolism*; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000235:genetics; D015203:Reproducibility of Results; D013997:Time Factors; D014026:Nicotiana / Q000235:genetics*
chemical_list: D017931:DNA Primers; D012313:RNA; D000594:Amino Acid Oxidoreductases; C021883:nopaline synthase