Jackson Colin J, Mochizuki Takashi, Barton Richard C
Institute of Biological Sciences, University of Wales, Aberystwyth, Wales, UK.
Department of Dermatology, Kanazawa Medical University, Uchinada, Japan.
J Med Microbiol. 2006 Oct;55(Pt 10):1349-1355. doi: 10.1099/jmm.0.46691-0.
The sequence of the nontranscribed spacer (NTS) region of the rDNA of Trichophyton mentagrophytes var. interdigitale strain 2111 was determined, and three individual subrepeat loci identified. The first repeat region contained eight tandem copies of a degenerate 33-43 bp sequence, whilst the second had two complete and two partial 300 bp repeats. The third locus contained six tandemly repetitive elements of between 67 and 89 bp, which showed sequence identity to the TrS2 repeats of Trichophyton rubrum. PCR amplification of the individual repetitive regions from 42 random isolates of T. mentagrophytes var. interdigitale identified fragment length polymorphisms at each locus. Sequence analysis of the PCR products revealed that the size variations resulted from differences in the copy number of each of the three sets of subrepeat elements, TmiS0, TmiS1 and TmiS2. In addition, some indels were present in the flanking regions of the TmiS1 repeats. Combining PCR fingerprints from each of the three polymorphic loci produced a total of 19 individual strain profiles. The method was rapid, reproducible and discriminatory, and the fragment patterns simple to interpret. PCR fingerprint analysis of variable tandem repeat loci in the T. mentagrophytes var. interdigitale NTS represents a valuable molecular typing method for future epidemiological investigations in this species.
测定了指间毛癣菌2111菌株核糖体DNA非转录间隔区(NTS)的序列,并鉴定出三个独立的亚重复位点。第一个重复区域包含一个33 - 43 bp简并序列的八个串联拷贝,而第二个区域有两个完整的和两个部分的300 bp重复序列。第三个位点包含六个67至89 bp的串联重复元件,与红色毛癣菌的TrS2重复序列具有序列同一性。从42株指间毛癣菌随机分离株中对各个重复区域进行PCR扩增,在每个位点鉴定出片段长度多态性。PCR产物的序列分析表明,大小变异是由三组亚重复元件TmiS0、TmiS1和TmiS2中每组拷贝数的差异引起的。此外,TmiS1重复序列的侧翼区域存在一些插入/缺失。结合三个多态性位点各自的PCR指纹图谱,共产生了19种菌株个体图谱。该方法快速、可重复且具有鉴别力,片段模式易于解读。指间毛癣菌NTS中可变串联重复位点的PCR指纹分析代表了一种有价值的分子分型方法,可用于该物种未来的流行病学调查。