Publication Abstract

Development of a real-time PCR assay for the identification of Gyrodactylus parasites infecting salmonids in northern Europe.

Development of a real-time PCR assay for the identification of Gyrodactylus parasites infecting salmonids in northern Europe.

C. M. Collins, R. Kerr*, R. Mcintosh and M. Snow.

Gyrodactylus salaris is a monogenean freshwater parasite which causes high mortality in wild Atlantic salmon and a number of countries employ monitoring programmes for its presence. A TaqMan®-MGB probe real-time multiplex assay targeting the internal transcribed spacer ribosomal DNA (ITS rDNA) was developed to simultaneously identify Gyrodactylus salaris/G. thymalli and two other commonly occurring Gyrodactylus species infecting salmonids in northern Europe: G. derjavinoides and G. truttae. In addition a Gyrodactylus genus level assay was developed to assess parasite DNA quality. The species specific real-time PCR method correctly identified target species from a wide geographical range and from a number of salmonid hosts. It did not amplify G. lucii nor G. teuchis. These species were successfully amplified using the Gyrodactylus genus real-time assay. The species specific real-time assay proved to be significantly faster than the currently employed molecular screening method of ITS rDNA PCR amplification followed by restriction fragment length polymorphism analyses (RFLP). However, as with ITS RFLP the real-time method did not distinguish between G. salaris and the non-pathogenic G. thymalli, it's principle advantage being a significant reduction in time to achieve an initial diagnostic screen before the employment of more in-depth analyses for those specimens giving a positive G. salaris/G. thymalli real-time result.

Reference:

C. M. Collins, R. Kerr*, R. Mcintosh and M. Snow. (2010) Development of a real-time PCR assay for the identification of Gyrodactylus parasites infecting salmonids in northern Europe. Diseases of Aquatic Organisms 90 (2) 135-142.

C. M. Collins, R. Kerr*, R. Mcintosh and M. Snow.

Diseases of Aquatic Organisms 90 (2) 135-142.