We report the construction of a genomic library and isolation of repetitive DNA probes from Cr. neoformans that are species- and variety-specific. The inserts of the genomic library were in a range of 0.3-1.8 kb, which were in normal distribution in length. Among 592 clones of the genomic library, there were 400(67.6%) clones inserted repetitive DNA sequence and 192(32.4%) inserted single copy DNA sequence of the Cr. neoformans genome. From the library, three specific probes were identified. Clone pCNIIA6 was serotype A-specific, pCNIIB5 species-specific, and pCNIIIG1 variety-specific (var. neoformans). On the basis of the high specificity and sensitivity of the signals observed by hybridization, we suggest that these probes provide a means for both biotyping and early diagnosis of Cr. neoformans.