Improvement of polymerase chain reaction-based Bt11 maize detection method by reduction of non-specific amplification.
The Bt11 maize-specific qualitative detection method based on polymerase chain reaction (PCR) in the JAS analytical test handbook has been widely used for administrative monitoring of GM crops and quality control of commercially distributed grains. In the present investigation, some apparently false-positive detections were observed in assays using the Bt11 maize-specific method, and these erroneous results were proved to have been caused by non-specific DNA amplification. We improved the detection method to reduce non-specific amplification by decreasing the concentration of magnesium ions in the PCR mixture. The subsequent evaluation of analytical performance demonstrated no marked difference between the currently used and the improved methods, except for the reduced non-specific amplification. We conclude that the currently used standard method should be replaced with the improved method for the reliable detection of Bt11 maize.
Mano
Junichi
J
National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
Yanaka
Yuka
Y
Akiyama
Hiroshi
H
Teshima
Reiko
R
Furui
Satoshi
S
Kitta
Kazumi
K
D002523:Edible Grain / Q000737:chemistry*; D005189:False Positive Reactions; D005504:Food Analysis / Q000379:methods*; D030841:Food, Genetically Modified; D016133:Polymerase Chain Reaction / Q000379:methods*