Differentiation of Naegleria fowleri and other naegleriae by polymerase chain reaction and hybridization methods

摘要: In order to detect and identify Naegleria fowleri strains an assay based on the Polymerase Chain Reaction (PCR) was evaluated. The amplified DNA fragments were detected by gel electrophoresis ethidium bromide staining, followed Southern blot hybridization with internal digoxigenin-labeled probe. A set of primers (B1B2) which flank a 678-bp region within virulence-associated gene, allowed for highly specific identification N. fowleri, since Naegleriae (N. lovaniensis, australiensis, gruberi, andersoni jadini) other Protozoa did not react. These amplification products from various organisms: Gram-positive bacteria, algae, y, yeasts human DNA. Whereas second (A1A2), different sequence, Acanthamoebae strains. After 40 cycles, limit detection single cell (cyst or trophozoite). Thus, PCR appears be rapid powerful tool fowleri.