Two percent ethidium bromide-stained agarose gel illustrating nested-PCR amplification of an 18S rRNA segment within C. parvum and genotype determination with VspI RFLP analysis. Approximately 200 ng of nested-PCR amplicon from singly or mixed genotypes was subjected to VspI digestion for 8 h at 37°C. Lane 1, genotype I amplicon; lane 2, genotype II amplicon; lane 3, digested genotype I amplicon illustrating 503- and 90-bp fragments; lane 4, digested genotype II; lane 5, digestion of both genotypes within a single reaction tube; lane M, 50-bp molecular weight marker (Gibco BRL, Grand Island, N.Y.).