Detection Methods for Biotech Cotton MON 15985 and MON 88913 by PCR

Abstract

Plants derived through agricultural biotechnology, or genetically modified organisms (GMOs), may affect human health and ecological environment. A living GMO is also called a living modified organism (LMO). Biotech cotton is a GMO in food or feed and also an LMO in the environment. Recently, two varieties of biotech cotton, MON 15985 and MON 88913, were developed by Monsanto Co. The detection method is an essential element for the GMO labeling system or LMO management of biotech plants. In this paper, two primer pairs and probes were designed for specific amplification of 116 and 120 bp PCR products from MON 15985 and MON 88913, respectively, with no amplification from any other biotech cotton. Limits of detection of the qualitative method were all 0.05% for MON 15985 and MON 88913. The quantitative method was developed using a TaqMan real-time PCR. A synthetic plasmid, as a reference molecule, was constructed from a taxon-specific DNA sequence of cotton and two construct-specific DNA sequences of MON 15985 and MON 88913. The quantitative method was validated using six samples that contained levels of biotech cotton mixed with conventional cotton ranging from 0.1 to 10.0%. As a result, the biases from the true value and the relative deviations were all within the range of +/-20%. Limits of quantitation of the quantitative method were all 0.1%. Consequently, it is reported that the proposed detection methods were applicable for qualitative and quantitative analyses for biotech cotton MON 15985 and MON 88913.

The Development and Standardization of Testing Methods for Genetically Modified Organisms and their Derived ProductsF

“…The reference molecules, usually the plasmids containing one or more functional fragments suitable for GM crop event detection, have been developed and proved to be a good alternative for CRM in GMO detection (Yang et al 2007; Zhang et al 2008). So far, over 20 reference molecules have been reported and used in the detection of several GM crops, for instance MON159855 and MON88913 cotton, GTS 40–3‐2 soybean, GT73 canola, MON863, GA21, TC1507, T25, MON810, NK603, Bt176, E‐3272, and 59122 maize (Taverniers et al 2005; Burns et al 2006; Lee et al 2007; Yang et al 2007; Zhang et al 2008; Li et al 2009; Shen et al 2010) etc.…”

Development of detection methods for six approved LM crops in Korea

“…또한, 제초제저항성 LM 옥수수 DAS-40278-9 (aad-1 유전자 삽입)와 LM 캐놀라 MON88302 (cp4 epsps 유전자 삽 입) PCR 반응 결과, 유전자변형 옥수수와 캐놀라에 대해서 만 특이적으로 도입유전자가 검출되는 것을 확인하였다 ( Fig. 3E and F Kim et al 2008;Kim et al 2015;Kim et al 2009b;Lee et al 2007;Min et al 2004;Moon et al 2012;Wu et al 2009;Yoo et al 2013 …”

Regulation and detection methods for genetically modified foods in Korea

“…A limited range of commercial reference materials is available for production of standard curves, so some researchers have produced their own calibration standards using purified genomic DNA or target DNA sequences cloned into plasmids [11,12]. Some Korean researchers have also investigated real-time PCR using plasmids as reference materials [13][14][15][16][17][18].…”