Analysis of the 5.8S rRNA and internal transcribed spacers regions of the variant Naegleria fowleri Thai strain

Abstract

The aim of this study was to analyze the internal transcribed spacers regions (ITS) of the of 5.8S ribosomal RNA (rRNA) gene of four isolants of Naegleria fowleri. Three of four Thai strains were isolated from patients and one from the environment. All four strains were confirmed to be N. fowleri by species-specific PCR using DNA extracted using a QIAamp DNA mini kit. The ITS lengths observed were ITS1, 85 bp; ITS2, 106 bp; and 5.8S, 176 bp. Five discriminating deca-nucleotide primers A1, A15, B10, B12, and B15 were used in this study. Specific prominent bands were observed after PCR with each primer: 600 bp with A1; 615 bp with A15; 1580 bp with B10; 930 and 510 bp with B12; and 310 bp with B15. All sequences were compared with the Japanese J16-1-42E sequence in the Genbank database. After alignment, our sequences contained only 0.5% variation from the J16-1-42 sequence. The ITS main products of the strain from the environment were similar to those of the three strains from Thai patients. The four Thai strains have essentially the same 5.8S rRNA genes as Cattenom Japanese J16(1) 42E strain. © 2007 Springer-Verlag.