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Quantification of Manila Clam Ruditapes philippinarum (Adams & Reeve, 1850) Larvae Based on SYBR Green Real-Time Polymerase Chain Reaction

2011; National Shellfisheries Association; Volume: 30; Issue: 3 Linguagem: Inglês

10.2983/035.030.0319

1943-6319

Javier Quinteiro, Pérez-Diéguez Lois, Ana Sánchez, Ricardo I. Pérez‐Martín, Cármen G. Sotelo, Manuel Rey‐Méndez,

Marine Bivalve and Aquaculture Studies

Biological and ecological research is seriously handicapped because of difficulties experienced in the reliable detection and quantification of bivalve larvae. This is a critical issue in the case of the Manila clam Ruditapes philippinarum (Adams & Reeve, 1850), a largely invasive and commercially relevant species, with important wild, cultured, and naturalized populations throughout the world. A SYBR Green real-time PCR assay, containing TPHI16S1F and TPHI16S2R primers (specific to R. philippinarum female mtDNA), was designed and tested to provide a rapid and high-throughput PCR-based method for larval quantification. Accurate estimations of larval numbers in spiked plankton samples point to the usefulness of this system. It can be used through a wide range of evaluated variable experimental conditions, such as the presence of closely related bivalve species, stationary-dependent plankton abundance, sampling volumes, and larval size.