Specific detection of N. fowleri by real-time PCR. The DNA was extracted from formalin-fixed brain samples which were stored for 17 months. (A) Real-time PCR amplification curves of DNA from cow and ewe samples demonstrating the presence of N. fowleri. (B) The corresponding melting curve profiles which were equivalent to that of N. fowleri. (∆) cow sample; (o) ewe sample; (x) negative control.