Improvement of Polymerase Chain Reaction-Based Bt11 Maize Detection Method by Reduction of Non-Specific Amplification

Abstract

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 di#erence 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.

Polymerase chain reaction: a powerful analytical tool in the field of food safety

“…GMO modifications have been introduced in most popular crops. Different PCR-based methods were developed to detect GMO material in cotton (Randhawa et al 2013), maize (Mano et al 2010), papaya (Yamaguchi et al 2006), pepper, tomato, eggplant, potato (Chaouachi et al 2008), rapeseed (canola) (Lee et al 2021), rice (Safaei et al 2019), soya bean (Park et al 2021), squash (Green et al 2004), sugar beet (Chaouachi et al 2013), watermelon (Bi et al 2019) and wheat (Terzi et al 2003). European Union Reference Laboratory for Genetically Modified Food and Feed (EURL GMFF) authorized few PCR assays for EU legal purposes (European Commission, GMO, 2023).…”