Detection of a genetic alteration and species identification of coho salmon (Oncorhynchus kisutch): a collaborative study

Abstract.

A PCR-based method for the identification of genetically modified coho salmon (Oncorhynchus kisutch) has been developed and evaluated by a collaborative study. The coho salmon contained an "all-salmon" gene-construct (OnMTGH1) consisting of the metallothionein-B-promoter fused to the full-length type-1 growth hormone gene, both from sockeye salmon (Oncorhynchus nerka).

Two PCRs were performed. The first PCR served as a control for amplifiable DNA present in the sample, and at the same time as a tool for species identification by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) test. For this PCR, universal primers located in the mitochondrial cytochrome b gene were used. The second PCR, working with primers located within the construct, detected the genetic alteration.

In a collaborative study with 15 laboratories, in each case the species and the genetic modification had been assigned correctly.