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Viral drug sensitivity testing using quantitative PCR: effect of tyrosine kinase inhibitors on polyomavirus BK
Parmjeet S Randhawa1, Noush A Farasati, Yuchen Huang
1Division of Transplant Pathology, University of Pittsburgh, Department of Pathology, E737 UPMC-Montefiore Hospital, 3459 Fifth Ave, Pittsburgh, PA 15213, USA.
Abstract:
Our objective was to determine whether quantitative polymerase chain reaction (PCR) can be used to measure the effect of tyrosine kinase (TK) inhibition on polyomavirus BK (BKV) replication. The BKV was grown in a cell culture system. The rate of viral replication in the presence or absence of the drug being tested was assessed by amplifying the viral genome using primers directed against the viral capsid 1 protein. Dasatinib, erlotinib, gefitinib, imatinib, sunitinib, and sorafenib all showed antiviral activity at micromolar concentrations. The 50% effective concentration for erlotinib and sorafenib was within blood concentrations readily achieved in human subjects. Quantitative PCR is a convenient method for viral drug sensitivity testing for slow-growing viruses that do not readily produce cytopathic effect. TK inhibitors deserve further consideration as a potential therapeutic option for BKV-associated nephropathy and hemorrhagic cystitis.
Insights
Quantitative PCR effectively measures the impact of tyrosine kinase (TK) inhibitors on polyomavirus BK (BKV) replication. Several TK inhibitors demonstrated antiviral activity, suggesting therapeutic potential for BKV-associated conditions.
Area of Science:
- Virology
- Molecular Biology
- Pharmacology
Background:
- Polyomavirus BK (BKV) is a significant cause of nephropathy and hemorrhagic cystitis, particularly in immunocompromised individuals.
- Current therapeutic options for BKV-associated diseases are limited, necessitating the exploration of novel treatment strategies.
- Tyrosine kinase (TK) inhibitors have shown promise in various antiviral applications.
Purpose of the Study:
- To evaluate the utility of quantitative polymerase chain reaction (qPCR) for assessing the antiviral effects of TK inhibitors on BKV replication.
- To determine the efficacy of specific TK inhibitors against BKV in a cell culture model.
Main Methods:
- BKV was cultured in vitro.
- Viral replication rates were quantified using qPCR by amplifying the BKV capsid 1 protein gene.
- The antiviral activity of dasatinib, erlotinib, gefitinib, imatinib, sunitinib, and sorafenib was assessed at micromolar concentrations.
Main Results:
- All tested TK inhibitors exhibited antiviral activity against BKV at micromolar concentrations.
- Erlotinib and sorafenib displayed a 50% effective concentration (EC50) achievable within human therapeutic blood levels.
- qPCR proved to be a practical method for drug sensitivity testing of slow-growing viruses like BKV.
Conclusions:
- Quantitative PCR is a viable and convenient method for assessing drug sensitivity in BKV replication.
- TK inhibitors demonstrate significant potential as a therapeutic avenue for managing BKV-associated nephropathy and hemorrhagic cystitis.
- Further investigation into TK inhibitors is warranted for BKV-related clinical conditions.
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