Identification of venvironmental isolates of acanthamoeba based on morphological and PCR techniques

Abstract

Acanthamoeba spp. are free-living amoebae that are widely distributed in the environment. Acanthamoeha spp. can also act as pathogens living in their host, and as free-living organisms in nature. In this study, water samples were collected at several locations around Universiti Malaysia Terengganu (UMT) to detect the presence of Acanthamoeba. These amoebae were identified by morphological and PCR techniques. Only Acanthamoeha were focused in this study. The microscopic observation of the cysts and tropozoites of Acanthamoeha was determined by following Page's description. To confirm the genus, primer pair JDP l and JDP2 was used to amplify nine region df3 of Acanthamoeba spp. which is highly specific to amplify the Acanthanioeba sp gene. The presence of 5 Bands after the PCR reaction in the gel indicates that the isolates are from genus Acanthamoeba. The bands were sent for sequencing analysis and the results showed that these isolates could be further confirmed up to the species level.