Development of a denaturing high-performance liquid chromatography method for detection of protist parasites of metazoans.

ABSTRACT: Increasingly, diseases of marine organisms are recognized as significant biotic factors affecting ecosystem health. However, the responsible disease agents are often unknown and the discovery and description of novel parasites most often rely on morphological descriptions made by highly trained specialists. Here, we describe a new approach for parasite discovery, utilizing denaturing high-performance liquid chromatography (DHPLC) reverse-phase ion-pairing technology. Systematic investigations of major DHPLC variables, including temperature, gradient conditions, and target amplicon characteristics were conducted to develop a mechanistic understanding of DNA fragment separation by DHPLC. As a model system, 18S rRNA genes from the blue crab (Callinectes sapidus) and a parasitic dinoflagellate

SUBMITTER: Troedsson C 

PROVIDER: S-EPMC2493188 | biostudies-literature | 2008 Jul

REPOSITORIES: biostudies-literature