To avoid misinterpretation of false positive results from laboratory contamination, a group of researchers designed and validated a set of synthetic positive controls for several aquatic invasive species.Follow our blog ! Subscribe to our RSS flux
In August 2015, Wilson et al. published a paper entitled “Recognizing false positives: synthetic oligonucleotide controls for environmental DNA surveillance” in the “Methods in ecology and Evolution” journal.
Summary: Environmental DNA (eDNA) is increasingly used for surveillance and detection of species of interest in aquatic and soil samples. A significant risk associated with eDNA methods is potential false positive results due to laboratory contamination. To minimize and quantify this risk, the authors of this paper designed and validated a set of synthetic oligonucleotides for use as species-specific positive PCR controls for several high-profile aquatic invasive species. The controls consist of species-specific sequences for the species of interest, with the addition of a synthetic insert containing recognition sites for several restriction enzymes. Following PCR, the presence of the synthetic insert can be detected using gel electrophoresis, restriction enzyme digests, or DNA sequencing. For quantitative PCR (qPCR), false positives in environmental samples can also be detected using a fluorescent probe designed to detect the synthetic insert. The generation of synthetic controls is a cost-effective, reproducible method that increases the power and reliability of eDNA testing by eliminating misinterpretation of false positive results from laboratory contamination.
Reference: Wilson, C.C., Wozney, K.M., Smith, C.M. (2015). Recognizing false positives: synthetic oligonucleotide controls for environmental DNA surveillance. Methods in Ecology and Evolution In press. doi.10.1111/2041-210X.12452.