Set-up and in-house validation of two multiplex event-specific PCR methods to simplify screening for GMOs without common screening elements

Since commercialization, the ever growing number of Genetically Modified (GM) crops make routine screening by European enforcement laboratories more expensive, labour-intensive and challenging every year. GM events, with elements for which no detection methods are available, further extend this screening because they require for event-specific methods to be included in the screening. Here, we describe the set-up and in-house validation of two tetraplex qPCR methods for simultaneous detection of six GM events that are not covered by common element-specific screening methods. The two qualitative multiplex PCR methods comprise previously developed and interlaboratory validated singleplex detection assays for GHB614 cotton, DP305423 soybean, CV127 soybean, DAS40278 maize, VCO1981 maize and 98140 maize. A plant-specific actin PCR was included in both multiplex PCR methods as a positive control. The validation data show that the assessed performance parameters are in accordance with the minimum performance requirements suggested by European Network of GMO Laboratories. The two multiplex PCR methods are reliable, ready to use and will lead to a faster and more efficient diagnostic routine GMO screening.