A real time genotyping PCR assay for polyomavirus BK

Gard, Lilli; Niesters, Hubert G. M.; Riezebos‐Brilman, Annelies

doi:10.1016/j.jviromet.2015.04.024

Cited by 13 publications

(19 citation statements)

References 28 publications

(34 reference statements)

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“…For example, Gard et al described a real-time PCR technique, but this technique did not determine subgroups (15). Matsuda et al developed a BKVgenotyping tool based on high-resolution melting analysis (16), but it was limited by its difficulty in distinguishing strains with the same polymorphisms.…”

Section: Discussionmentioning

confidence: 99%

A Simple and Reliable Strategy for BK Virus Subtyping and Subgrouping

et al. 2017

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BK virus (BKV)-associated diseases in transplant recipients are an emerging issue. However, identification of the various BK virus subtypes/subgroups is a long and delicate process on the basis of currently available data. Therefore, we wanted to define a simple and effective one-step strategy for characterizing all BK virus strains from the VP1 gene sequence. Based on the analysis of 199 available complete DNA VP1 sequences, phylogenetic trees, alignments, and isolated polymorphisms were used to define an effective strategy for distinguishing the 12 different BK virus subtypes/subgroups. Based on the 12 subtypes identified from the 199 complete BKV VP1 sequences (1,089 bp), 60 mutations that can be used to differentiate these various subtypes/subgroups were identified. Some genomic areas were more variable and comprised mutational hot spots. From a subregion of only 100 bp in the VP1 region (1977 through 2076), we therefore constructed an algorithm that enabled rapid determination of all BKV subtypes/subgroups with 99% agreement (197/199) relative to the complete VP1 sequence. We called this domain of the BK viral genome the BK typing and grouping region (BKTGR). Finally, we validated our viral subtype identification process in a population of 100 transplant recipients with 100% efficiency. The new simpler method of BKV subtyping/subgrouping reported here constitutes a useful tool for future studies that will help us to more clearly understand the impact of BKV subtypes/subgroups on diagnosis, infection, and BK virus-associated diseases.

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Section: Discussionmentioning

confidence: 99%

A Simple and Reliable Strategy for BK Virus Subtyping and Subgrouping

et al. 2017

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“…Reported prevalence percentages are generally based on BKPyV isolates from viruric and viremic (immunocompromised) individuals, and therefore may not represent the BKPyV genotype distribution in the general (immunocompetent) population. Coinfection of a dominant genotype with other BKPyV genotypes/subtypes (quasispecies) is common [24, 28, 29].…”

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confidence: 99%

Source and Relevance of the BK Polyomavirus Genotype for Infection After Kidney Transplantation

Wunderink

Brouwer

Gard

et al. 2019

Open Forum Infectious Diseases

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Background BK polyomavirus (BKPyV)–associated nephropathy (BKPyVAN) is a major threat for kidney transplant recipients (KTRs). The role of specific BKPyV genotypes/serotypes in development of BKPyVAN is poorly understood. Pretransplantation serotyping of kidney donors and recipients and posttransplantation genotyping of viremic recipients, could reveal the clinical relevance of specific BKPyV variants. Methods A retrospective cohort of 386 living kidney donor-recipient pairs was serotyped before transplantation against BKPyV genotype I–IV viral capsid protein 1 antigen, using a novel BKPyV serotyping assay. Replicating BKPyV isolates in viremic KTRs after transplantation were genotyped using real-time polymerase chain reaction and confirmed by means of sequencing. BKPyV serotype and genotype data were used to determine the source of infection and analyze the risk of viremia and BKPyVAN. Results Donor and recipient BKPyV genotype and serotype distribution was dominated by genotype I (>80%), especially Ib, over II, III and IV. Donor serotype was significantly correlated with the replicating genotype in viremic KTRs (P < .001). Individual donor and recipient serotype, serotype (mis)matching and the recipient replicating BKPyV genotype were not associated with development of viremia or BKPyVAN after transplantation. Conclusions BKPyV donor and recipient serotyping and genotyping indicates the donor origin of replicating BKPyV in viremic KTRs but provides no evidence for BKPyV genotype–specific virulence.

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“…Samples collected before January 2012 were extracted using the MagNaPure LC (Roche Diagnostics, Germany) and BKPyV-DNA was measured with RT-PCR(, , ). After January 2012 samples were extracted and amplified as previously described by Gard et al[13]. Both RT-PCR’s were a multiplex with the internal control, seal herpes virus (PhHV) (, , ).…”

Section: Methodsmentioning

confidence: 99%

“…The BKPyV main genotype was determined from the first BKPyV positive sample in a patient using the RT-PCR as described previously [13]. …”

Section: Methodsmentioning

confidence: 99%

A delicate balance between rejection and BK polyomavirus associated nephropathy; A retrospective cohort study in renal transplant recipients

et al. 2017

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BackgroundThe immunosuppressive agents mycophenolate acid (MPA) and tacrolimus (Tac) are associated with a higher incidence of BK polyomavirus nephropathy (BKPyVAN).In this observational retrospective cohort study, the frequency of BK polyomavirus (BKPyV) complications over a 24-month period was studied.Methods358 renal transplant recipients (RTR) treated with MPA, with either cyclosporine A (CsA) (CsAM group) or Tac (TacM group) and mostly prednisolone, were included.ResultsIncidence of BKPyV-viremia was not significantly different between the CsAM (n = 42/191) (22.0%) and the TacM (n = 36/167) (21.6%) group. Biopsy proven BKPyVAN occurred more often in the TacM group (6.6%) versus the CsAM group (2.1%) (p = 0.03). Longitudinal data analysis showed a significant earlier decline of viral load in plasma in the CsAM group compared to the TacM group (p = 0.005).The incidence of biopsy proven acute rejection (BPAR) was significantly higher in the CsAM (19.9%) compared to the TacM (10.8%) (p = 0.02) group. Graft loss, estimated glomerular filtration rate and mortality rate did not differ in both treatment groups.ConclusionIn conclusion, this study shows that immunosuppressive treatment with Tac and MPA compared to CsA and MPA is associated with a lower incidence of BPAR, but at the cost of an increased risk of developing BKPyVAN in the first two years post-transplant.

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A real time genotyping PCR assay for polyomavirus BK

Cited by 13 publications

References 28 publications

A Simple and Reliable Strategy for BK Virus Subtyping and Subgrouping

A Simple and Reliable Strategy for BK Virus Subtyping and Subgrouping

Source and Relevance of the BK Polyomavirus Genotype for Infection After Kidney Transplantation

A delicate balance between rejection and BK polyomavirus associated nephropathy; A retrospective cohort study in renal transplant recipients

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