Katsu Masakazu, Kidd Sarah, Ando Akikazu, Moretti-Branchini Maria Luiza, Mikami Yuzuru, Nishimura Kazuko, Meyer Wieland
Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University, Chuo-ku, Chiba-city, Chiba 260-8673, Japan.
FEMS Yeast Res. 2004 Jan;4(4-5):377-88. doi: 10.1016/S1567-1356(03)00176-4.
Sequences of the internal transcribed spacer (ITS) region including the 5.8S rRNA gene delineated seven genotypes within the three varieties of Cryptococcus neoformans via specific combinations of eight nucleotide differences located at positions 10, 11, 15, 19, 108 (ITS1), 221 (5.8S), 298 and 346 (ITS2). The ITS types correlated to polymerase chain reaction fingerprint/random amplification of polymorphic DNA (RAPD) molecular types: with ITS type 1 (ATACTAGC)=C. neoformans var. grubii, molecular types VNI+VNII and the serotype A allele of the AD hybrid, VNIIIA; ITS type 2 (ATATAGGC)=the serotype D allele of the AD hybrid, VNIIIB, and C. neoformans var. neoformans, VNIV; and ITS type 3 (GCGCTGGC) and ITS type 7 (ACGCTGGC)=VGI=RAPD type III, ITS type 4 (ACACTGAC)=VGII=RAPD type II, ITS type 5: (ACACTGGG)=VGIII=RAPD type I, ITS type 6 (ACACTGGC)=VGIV=RAPD type IV, all corresponding to C. neoformans var. gattii. Cloned sequences from serotype AD revealed that the hybrid serotype is diploid at the ITS1-5.8S-ITS2 locus carrying the ITS type 1 (ATACTAGC) and the ITS type 2 (ATATAGGC) alleles. ITS sequencing is a useful technique for genotyping the three C. neoformans varieties and for subtyping within C. neoformans var. gattii.
内部转录间隔区（ITS）序列，包括5.8S rRNA基因，通过位于第10、11、15、19、108位（ITS1）、221位（5.8S）、298位和346位（ITS2）的8个核苷酸差异的特定组合，在新型隐球菌的三个变种中划分出了7种基因型。ITS类型与聚合酶链反应指纹/随机扩增多态性DNA（RAPD）分子类型相关：ITS类型1（ATACTAGC）=新型隐球菌格鲁比变种，分子类型VNI+VNII以及AD杂交种的血清型A等位基因，VNIIIA；ITS类型2（ATATAGGC）=AD杂交种的血清型D等位基因，VNIIIB，以及新型隐球菌新型变种，VNIV；ITS类型3（GCGCTGGC）和ITS类型7（ACGCTGGC）=VGI=RAPD类型III，ITS类型4（ACACTGAC）=VGII=RAPD类型II，ITS类型5：（ACACTGGG）=VGIII=RAPD类型I，ITS类型6（ACACTGGC）=VGIV=RAPD类型IV，所有这些都对应于新型隐球菌格特变种。来自血清型AD的克隆序列显示，该杂交血清型在ITS1-5.8S-ITS2位点是二倍体，携带ITS类型1（ATACTAGC）和ITS类型2（ATATAGGC）等位基因。ITS测序是对新型隐球菌的三个变种进行基因分型以及对新型隐球菌格特变种进行亚型分型的有用技术。
