FIG. 1.

Molecular identification of Cyclospora and Eimeria species. (A) Conventional nested-PCR amplification. Partially purified oocysts (100 to 1,000) were spotted onto FTA filters that were used as a PCR template. For the primate-derived Cyclospora-like organisms, 2 μl of purified DNA was used as a template. A primary amplicon (636 bp [not shown]) was generated from each of these templates with the F1E-R2B primer pair, and 1 μl of this product was used in a nested amplification with the F3E-R4B primer pair to generate the 294-bp amplicon shown. (B) RFLP analysis of nested-PCR amplicons. Nested-PCR amplicons were digested with MnlI and analyzed by gel electrophoresis with 5% NuSieve 3:1 agarose containing ethidium bromide (0.2 μg/ml).