2.4 DNA Sequence Analyses
PCR products were prepared for sequencing using a QIAquick Gel Extraction Kit (Qiagen). DNA sequencing of the purified PCR products was performed using a model ABI3730XL sequencer with a BigDye® Terminator v3.1 Cycle Sequencing Kit (ABI, Foster City, CA, USA). Viral MCPyV genotypes were identified by direct sequence analysis. The obtained DNA sequence data were BLAST searched at the National Center for Biotechnology Information (NCBI) (http://www.ncbi.nlm.nih.gov/blast/Blast.cgi). All MCPyV sequence alignments were performed using BioEdit version 7.1.9 (www.mbio.ncsu.edu/bioedit/bioedit.html) and compared with the six reference sequences of North American (MCC350, EU375803.1, MCC339, and EU375804.1), Japanese (TKS and FJ 464337), Swedish, French, Italian (MKL-1 and FJ173815), Chinese (HB039C and KC571692.1) and Korean (KIB and MK561422.1) MCPyV isolates.12,31-34