DNA amplified from individual Plasmodium vivax oocysts, produced by feeding mosquitoes directly on naturally infected humans in Thailand, was used to study cross-mating of 2 polymorphs of the circumsporozoite (CS) gene, VK 210 and VK 247. Alleles were detected in matched blood parasites, sporozoites, and individual oocysts with oligoprobes specific to characteristic repeat units. Oocysts developing from 3 cases in which mixed alleles were present in the blood parasites had genotype frequencies, including hybrids, consistent with the Hardy-Weinberg equilibrium. There was apparently no barrier to hybridization of the 2 alleles nor a bias, as has been found in some laboratory experiments, favoring hybrid formation. These are the first measurements of cross-mating frequencies directly from natural Plasmodium infections and the first observations of genetic hybridization in P. vivax.