Rapid Identification of Rumen Protozoa by Restriction Analysis of Amplified 18S rRNA Gene

Abstract

A rapid method has been developed for molecular identification of rumen ciliates without the need for cultivation. Total DNA was isolated from single protozoal cells by the Chelex method and nearly complete protozoal 18S rRNA genes were amplified and subjected to restriction fragment length polymorphism analysis. On the basis of restriction patterns generated a molecular key was elaborated allowing identification of protozoa solely by a molecular technique without prior knowledge of morphology. No differences were observed between identical species originating from different animals or geographic locations, or between morphological variants of the same species. The ARDREA analysis described here provides a rapid and convenient way for identification and diversity studies of rumen protozoa.