학술논문

Laboratory-performance Study of Quantitative PCR Methods to Analyze an Approved Genetically Modified Maize (Mon810 Line) / 遺伝子組換えトウモロコシ(Mon810系統)の定量PCR法を対象とした外部精度管理試験
Document Type
Journal Article
Source
食品衛生学雑誌 / Food Hygiene and Safety Science (Shokuhin Eiseigaku Zasshi). 2006, 47(1):15
Subject
DNA extraction method
DNA抽出法
PCR
detection method
genetically modified maize
laboratory-performance study
ポリメラーゼ連鎖反応
外部精度管理
検査方法
遺伝子組換えトウモロコシ
Language
Japanese
ISSN
0015-6426
1882-1006
Abstract
A laboratory-performance study was carried out to investigate factors affecting the reliability of the quantitative PCR method to analyze an approved genetically modified (GM) maize (Mon810 line).Test maize powdered samples were prepared as blind samples containing a high (assigned value; 5.45%) or low (assigned value; 0.35%) concentration of the Mon810 line. After confirmation of their homogeneity, they were provided to 27 laboratories participating in the collaborative study. The data were collected from all laboratories and statistically analyzed. Two laboratories, which used a Roche LightCycler (LC), reported significantly high test values. A further examination showed that the LC method is greatly affected by the equipment itself or PCR reagents, resulting in poor repeatability. On the other hand, some laboratories, which used ABI quantitative PCR equipment, reported erroneous test values. In these laboratories, the errors appeared to have been due to inadequate quality and/or yield of DNA. To identify factors affecting the test values, analysis of the measured values for the taxon-specific gene will be useful. Furthermore, the modified silica-gel membrane DNA extraction method made it possible to extract the required amounts of DNA more easily and in a shorter time than before.