Quantitative analysis of herpesvirus sequences from normal tissue and fibropapillomas of marine turtles with real-time PCR

Quantitative real-time PCR has been used to measure fibropapilloma-associat ed turtle herpesvirus (FPTHV) pol DNA loads in fibropapillomas, fibromas, a nd uninvolved tissues of green, loggerhead, and olive ridley turtles from H awaii, Florida, Costa Rica, Australia, Mexico, and the West Indies. The vir al DNA loads from tumors obtained from terminal animals were relatively hom ogenous (range 2-20 copies/cell), whereas DNA copy numbers from biopsied tu mors and skin of otherwise healthy turtles displayed a wide variation (rang e 0.001-170 copies/cell) and may reflect the stage of tumor development. FP THV DNA loads in tumors were 2.5-4.5 logs higher than in uninvolved skin fr om the same animal regardless of geographic location, further implying a ro le for FPTHV in the etiology of fibropapillomatosis. Although FPTHV pol seq uences amplified from tumors are highly related to each other, single signa ture amino acid substitutions distinguish the Australia/Hawaii, Mexico/Cost a Rica, and Florida/Caribbean groups. (C) 2001 Academic Press.