PCR fingerprinting of Trichophyton mentagrophytes var. interdigitale using polymorphic subrepeat loci in the rDNA nontranscribed spacer.
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.
Jackson
Colin J
CJ
Institute of Biological Sciences, University of Wales, Aberystwyth, Wales, UK.
Mochizuki
Takashi
T
Department of Dermatology, Kanazawa Medical University, Uchinada, Japan.
Barton
Richard C
RC
Mycology Reference Centre, Department of Microbiology, General Infirmary, and University of Leeds, Leeds, UK.
D001530:Belgium; D016172:DNA Fingerprinting / Q000379:methods*; D004271:DNA, Fungal / Q000235:genetics*; D021901:DNA, Intergenic; D004274:DNA, Recombinant / Q000235:genetics; D006801:Humans; D007194:India; D007564:Japan; D008969:Molecular Sequence Data; D016533:Mycological Typing Techniques / Q000379:methods; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D013045:Species Specificity; D020080:Tandem Repeat Sequences; D014005:Tinea / Q000382:microbiology*; D014249:Trichophyton / Q000145:classification* / Q000235:genetics; D006113:United Kingdom
D004271:DNA, Fungal; D021901:DNA, Intergenic; D004274:DNA, Recombinant