A consensus real-time RT-PCR for detection of all genotypic variants of yellow head virus of penaeid shrimp.

Wijegoonawardane, Priyanjalie K M; Cowley, Jeff A; Walker, Peter J

A real-time quantitative (q)RT-PCR employing consensus degenerate PCR primers was developed to detect all six genotypes known currently to comprise the yellow head virus (YHV) complex and found commonly in Penaeus monodon shrimp. The test primers possess only limited (eight-fold) degeneracy and target ORF1b gene sequences identified to be highly conserved amongst 57 strains of the six genotypes detected in P. monodon sourced from various regions of the Indo-Pacific. The qRT-PCR amplifies a 147bp sequence and analysis of dilutions of synthetic genotype 2 RNA showed it to be 99.8% efficient and capable of detecting as few as 2.5 RNA copies reliably. As the test detects all six YH-complex genotypes, is extremely sensitive, capable of quantifying infection loads, and amenable to high-throughput application, it should prove useful for managing infections in P. monodon broodstock and seedstock used for aquaculture.