Wu J, Liao W, Chai J
Department of Dermatology, Shanghai Changzheng Hospital, P. R. China.
Mycoses. 1997 Dec;40(9-10):385-9. doi: 10.1111/j.1439-0507.1997.tb00256.x.
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.
我们报道了新型隐球菌基因组文库的构建以及从该菌中分离出的具有种属和变种特异性的重复DNA探针。基因组文库的插入片段大小在0.3 - 1.8 kb之间，长度呈正态分布。在基因组文库的592个克隆中，有400个（67.6%）克隆插入了重复DNA序列，192个（32.4%）克隆插入了新型隐球菌基因组的单拷贝DNA序列。从文库中鉴定出了三种特异性探针。克隆pCNIIA6是A血清型特异性的，pCNIIB5是种特异性的，pCNIIIG1是变种特异性的（新生变种）。基于杂交观察到的信号具有高特异性和高敏感性，我们认为这些探针为新型隐球菌的生物分型和早期诊断提供了一种方法。
