From: Direct Identification of Dermatophyte DNA From Clinical Specimens by a Nested Polymerase Chain
Reaction Assay
Arch Dermatol. 2007;143(6):799-816. doi:10.1001/archderm.143.6.799
Figure Legend:
Representation of the ribosomal DNA (rDNA) gene complex in fungi denoting gene order and the position of the internal transcribed
spacer (ITS). Primers were designed on this complex and were designated as ITS1, ITS2, ITS3, and ITS4 as their locations on
genes. Oligonucleotide primers ITS1 (5′-TCC GTA GGT GAA CCT GCG G-3′) and ITS3 (5′-GGA AGT AAA AGT CGT AAC AAG G3′) were both designed on 18S rDNA. The primer ITS2 (5′-GCT GCG TTC TTC ATC GAT GC-3′) was designed on 5.8S rDNA, and
ITS4 (5′-TCC TCC GCT TAT TGA TAT GC-3′) was
designed
on 28S
rDNA.Medical
Oligonucleotide primer pairs ITS3 and ITS4 were used in
Copyright
© 2007
American
Date
of download:
7/31/2017chain reaction (PCR) amplification reaction. Then the templates of nested PCR were the products of firstthe first-round
polymerase
Association. All rights reserved.
round PCR using ITS1 and ITS2 as paired primers.
From: Direct Identification of Dermatophyte DNA From Clinical Specimens by a Nested Polymerase Chain
Reaction Assay
Arch Dermatol. 2007;143(6):799-816. doi:10.1001/archderm.143.6.799
Figure Legend:
Genomic DNA samples extracted from 5 different skin scrapings were amplified by a MagNA pure Light Cycler System (Roche
Diagnostics GmbH, Mannheim, Germany) with nested polymerase chain reaction (PCR) and analyzed by 2% agarose gel
electrophoresis. The left panel represents the first round of PCR, while the right panel represents the second round. L and L′
represent the 100–base pair (bp) DNA ladder; 1 and 1′ through 5 and 5′, 5 different skin scraping tests; 6 and 6′, negative controls
(Staphylococcus epidermidis genome); and 7 and
7′, positive
controls
(Trichophyton
mentagrophytes genome). All of the
Copyright
© 2007
American
Medical
Date
of download:
7/31/2017
distinguishable
molecular
weights of first PCR products
were
about
750
bp
(lanes
5
and
7); nested PCR products, 350 bp (lanes 1′Association. All rights reserved.
5′ and 7′). These 5 clinical specimens belong to the genera Trichophyton and Microsporum and were proved by DNA sequencing.