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1.
Article in English | MEDLINE | ID: mdl-30231168

ABSTRACT

BKV and JCV belong to the Polyomaviridae family and are opportunistic agents associated with complications in immunocompromised individuals. Although a single screening assay for both viruses would be convenient, the diversity of BKV and JCV serotypes and genotypes is a methodological challenge. 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. Specific products were observed when JCV and BKV plasmids were mixed in the same reaction. In field sample testing, the duplex snPCR detected and distinguished both viruses in different biological samples. Results were confirmed by Sanger's sequencing. The geographical complexity of BKV and JCV serotypes and genotypes imposes limits to a simple and universal method that could detect each virus. However, we describe here a sensitive and reliable PCR technique for BKV and JCV diagnosis that overcomes these limitations and could be universally applied.


Subject(s)
BK Virus/isolation & purification , DNA, Viral/genetics , JC Virus/isolation & purification , Polymerase Chain Reaction/methods , BK Virus/classification , BK Virus/genetics , Genotype , Humans , JC Virus/classification , JC Virus/genetics
2.
Rev Inst Med Trop Sao Paulo ; 58: 58, 2016 07 11.
Article in English | MEDLINE | ID: mdl-27410918

ABSTRACT

The aim of this study was to investigate the association of EBV and HPV with gingivitis and/or periodontitis according to the immunologic status. To this end, 74 oral biopsies from transplanted and non-transplanted individuals with the abovementioned oral manifestations were submitted to a screening by PCR for both viruses. According to the results, EBV was strongly associated with gingivitis and/or periodontitis in transplanted individuals (p = 0.011) but not HPV (p = 0.766). EBV-HPV co-detections did not enhance the presence of tissue injury as well. Although a causal relationship was not investigated in this study, the higher frequency of these two oncoviruses in lesion tissues must be investigated in follow-up studies, especially among immunocompromised individuals.


Subject(s)
Epstein-Barr Virus Infections/diagnosis , Gingivitis/virology , Kidney Transplantation , Papillomavirus Infections/diagnosis , Periodontitis/virology , Case-Control Studies , DNA, Viral/genetics , Gingivitis/diagnosis , Herpesvirus 4, Human/genetics , Herpesvirus 4, Human/isolation & purification , Humans , Papillomaviridae/genetics , Papillomaviridae/isolation & purification , Periodontitis/diagnosis , Polymerase Chain Reaction
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