PCR techniques applied to Hematodinium spp. and Hematodinium-like dinoflagellates in decapod crustaceans

Abstract

The amount of DNA recovered from 1 X 106 cells of Hematodinium-like dinoflagellates from Chionoecetes bairdi was 75 ng. PCR (polymerase chain reaction) amplification products were created from the host and Hematodinium spp. Amplified products of 680 base pairs (bp) were produced from Hematodinium-like dinoflagellates from C, bairdi for DNA concentrations of 100, 50, 20, 10 and 1 ng pl-' This amplified fragment was produced only from host tlssue infected with the parasite. The lowest detectable limit of 1 ng of DNA from Hematodinium-like sp. from C. bairdi corresponded to 1.0 X 104 cells. Diagnostic bands of 680 bp were produced from Hematodinjum spp. and Hematodinium-like spp. isolated from the following hosts: Chionoecetes op~lio, Callinectes sapidus, Nephrops norvegicus and Portunus pelagicus. Host DNA (C. bairdi) produced a faint band of 1300 bp for concentrations of 100, 50 and 20 ng pl-'.