Collaborative Validation of an Event-Specific Quantitative Real-Time PCR Method for Genetically Modified Rice Event TT51‑1 Detection

In
this study, a collaborative trial of validating a real-time PCR method
for the TT51-1 rice event was organized, including six participating
laboratories. In this validation, serially diluted solutions from
homogeneous genomic DNA of the TT51-1 event were used to construct
standard curves of the TT51-1 event and phospholipase D (PLD) assays. The PCR efficiency was 95%, and the R2 coefficient was 0.99 for the TT51-1 system. The mean quantitative
values for blind samples containing 0.1%, 0.5% 1%, 5%, and 10% (w/w)
TT51-1 corresponded to 0.1%, 0.51%, 1.06%, 4.83%, and 9.62%, respectively,
with a bias (%) ranging from −3.77% to 5.87%. The repeatability
and reproducibility were all below 25% across the entire dynamic range.
Furthermore, the measurement uncertainties of the quantitative results
were estimated to be 0.10%, 0.20%, 0.40%, 1.76%, and 3.52% (w/w) for
the tested samples. Both the LOD and LOQ were calculated to be 0.22%.
This collaborative trial demonstrated that the TT51-1 method produces
reliable, comparable, and reproducible results for a given sample
set and can be adopted as a detection standard for testing laboratories.