A globally applicable PCR-based detection and discrimination of BK and JC polyomaviruses.BKV and JCV belong to the Polyomaviridae family and are opportunistic agents associated with complications in immunocompromised individuals.  In this paper, we developed a PCR method able to detect and segregate BKV and JCV, despite these genetic discrepancies.  A duplex semi-nested PCR (duplex snPCR) was designed to target a conserved region (639nt-1516nt) within the VP2 gene.  In the first PCR, a primer set common to all BKV and JCV serotypes/ genotypes was used, followed by a semi-nested PCR with internal primers for BKV and JCV segregation.  The limit of detection of the duplex snPCR was as low as 10 copies of BKV or JCV plasmids/μL.  In field sample testing, the duplex snPCR detected and distinguished both viruses in different biological samples.  However, we describe here a sensitive and reliable PCR technique for BKV and JCV diagnosis that overcomes these limitations and could be universally applied.
PCR
A duplex semi-nested PCR (duplex snPCR) was designed to target a conserved region (639nt-1516nt) within the VP2 gene.  The limit of detection of the duplex snPCR was as low as 10 copies of BKV or JCV plasmids/μL.  In field sample testing, the duplex snPCR detected and distinguished both viruses in different biological samples. 
duplex snPCR
Results were confirmed by Sanger's sequencing. 
Sanger's sequencing