1
Formalin-fixed, paraffin-embedded (FFPE) skin biopsy specimens
Used as samples from patients for the evaluation of CuV status
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2
WaxFree DNA Extraction kit (TrimGen, Sparks, MD, USA)
For DNA extraction from archived FFPE tissues
3
Microtome blades
Changed between each embedded block to prevent cross-contamination between specimens
4
Blank extraction control (reagent only)
Included for each DNA extraction step as a control
5
Real-time TaqMan-based quantitative polymerase chain reaction (qPCR)
For detection and quantification of CuV DNA
6
Primers and probes
To amplify the gene encoding the CuV viral capsid protein 1 (VP1) and VP2
7
Negative DNA extraction controls and molecular biology-grade water
Used as no-template controls to ensure negative control conditions in qPCR
8
RNase P
Used as a housekeeping gene to control for the quality of the DNAs and PCR reactions
9
CuV VP1/VP2 PCR product
Used as the standard for the qPCR assay
10
Clustal W
For nucleotide sequence alignment
11
PCR
To amplify CuV VP1/VP2 sequences for phylogenetic analysis
12
Maximum-likelihood method in MEGA X
To construct phylogenetic trees
13
Nucleotide BLAST program
For nucleotide identity analysis