Age-Specific Seroprevalence of Merkel Cell Polyomavirus, BK Virus, and JC Virus

ABSTRACTWe produced capsids of Merkel cell polyomavirus (MCPyV) in a baculovirus expression system and developed a virus-like particle (VLP) enzyme-linked immunosorbent assay (ELISA). To determine age-specific seroprevalence, serum samples were collected from 947 individuals attending hospital outpatient clinics and ranging in age from 1 to 93 years. To evaluate the association between exposure to MCPyV and Merkel cell cancer (MCC), plasma samples were obtained from 33 MCC patients and 37 controls. MCPyV seroprevalence was 45% in children under 10 years of age, increased to 60% in the next decade of life, and peaked at 81% among those 60 to 69 years of age. Levels of MCPyV capsid antibodies were positively correlated with age (P= 0.007). Virus specificity of MCPyV seroreactivity was supported by competitive inhibition of reactivity by MCPyV VLPs and not by BK polyomavirus (BKPyV) VLPs. MCPyV seroprevalence was greater among MCC patients (91%) than controls (68%; age-adjustedPvalue, 0.32); the mean level of MCPyV antibodies was also greater (P= 0.04). The age-specific seroprevalence of MCPyV shares with previously known polyomaviruses, BKPyV and JC polyomavirus (JCPyV), evidence of widespread exposure in human populations beginning early in life. MCPyV age-specific seroprevalence also has unique features. Seroprevalence among children is higher than that of JCPyV but lower than that of BKPyV. Among older adults, MCPyV seroprevalence remains high, while that of BKPyV declines and that of JCPyV continues to rise. In agreement with results from other studies, we found an association between MCPyV seropositivity and MCC, and higher levels of serum MCPyV capsid antibodies in MCC patients than in controls.

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Immunoglobulin G, A, and M Responses to BK Virus in Renal Transplantation

Clinical and Vaccine Immunology ◽

10.1128/cvi.00114-06 ◽

2006 ◽

Vol 13 (9) ◽

pp. 1057-1063 ◽

Cited By ~ 40

Author(s):

Parmjeet S. Randhawa ◽

Gaurav Gupta ◽

Abhay Vats ◽

Ron Shapiro ◽

Raphael P. Viscidi

Keyword(s):

Viral Replication ◽

Optical Density ◽

Immunoglobulin G ◽

Jc Virus ◽

Bk Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Kidney Transplant Recipients ◽

Serum Samples ◽

Igm Antibodies ◽

Active Viral Replication

ABSTRACT Immunoglobulin G (IgG), IgA, and IgM antibodies were measured in serum samples from 71 organ donors, 81 kidney transplant recipients at transplantation, and 67 patients during the posttransplant period by using a virus-like particle-based enzyme-linked immunosorbent assay (ELISA). BK virus (BKV) and JC virus DNA were detected in urine and plasma by real-time PCR. IgG antibodies to BKV were demonstrated in the majority (80.3 to 100%) of patients irrespective of clinical category, but titers were highest in patients with active viral replication. IgA antibodies were present with greater frequency (72.7 to 81.3% versus 0 to 23.6%; P < 0.001) and higher titer (mean optical density, 0.11 to 0.15 versus 0.05 to 0.08; P < 0.001) in patients who were BKV DNA positive than those who were BKV DNA negative. IgM antibodies showed a similar pattern of reactivity but lower frequency in the setting of active viral replication (9.1 to 43.7% versus 0 to 1.4%; P < 0.001). A rise in IgG level of >0.577 optical density (OD) units or a rise in IgA or IgM level of >0.041 OD units was strongly associated with active viral replication. Urine viral load showed a positive correlation with IgM titer (r = 0.22) but a negative correlation with IgG titer (r = −0.28) and IgA titer (r = −0.1). Chronic dialysis patients typically did not have serologic or virologic evidence of active BKV infection. Anti-BKV titers did not rise in patients with JC viruria. In conclusion, measurement of anti-BKV antibody titer and class response can be used to detect the onset of viral replication. ELISAs can be quite specific despite considerable sequence homology between BK virus and JC virus.

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Pestivirus infections in cervids from the Czech Republic

Veterinární Medicína ◽

10.17221/29/2009-vetmed ◽

2009 ◽

Vol 54 (No. 4) ◽

pp. 191-193

Author(s):

K. Sedlak ◽

T. Girma ◽

J. Holejsovsky

Keyword(s):

Czech Republic ◽

Cervus Elaphus ◽

Red Deer ◽

Competitive Inhibition ◽

Enzyme Linked Immunosorbent Assay ◽

Border Disease Virus ◽

Serum Samples ◽

The Czech Republic ◽

Diarrhea Virus ◽

Border Disease

372 sera of cervids from the Czech Republic were examined for antibodies to the bovine viral diarrhea virus (BVDV) and border disease virus (BDV) by competitive-inhibition enzyme-linked immunosorbent assay (ELISA), and for the presence of the BVDV by AgELISA. Antibodies to BVDV/BDV were found in 0.6% (two positive/305 tested) red deer (Cervus elaphus). BVDV/BDV antibodies were not found in four sika deer (Cervus Nippon) and 63 fallow deer (Dama dama). All serum samples were BVDV antigen negative. Our results confirmed that red deer in the Czech Republic are only rarely infected with Pestiviruses. This was the first survey of pestiviruses in farmed and wild cervids in the Czech Republic.

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Merkel cell polyomavirus detection in Merkel cell cancer tumors in Northern Germany using PCR and protein expression

Journal of Medical Virology ◽

10.1002/jmv.23808 ◽

2013 ◽

Vol 86 (10) ◽

pp. 1813-1819 ◽

Cited By ~ 6

Author(s):

Miriam Leitz ◽

Kristin Stieler ◽

Adam Grundhoff ◽

Ingrid Moll ◽

Johanna M. Brandner ◽

...

Keyword(s):

Protein Expression ◽

Cell Cancer ◽

Merkel Cell Polyomavirus ◽

Merkel Cell ◽

Northern Germany

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The effect of plasma exchange on serum anti-JC virus antibodies

Multiple Sclerosis Journal ◽

10.1177/1352458512467502 ◽

2012 ◽

Vol 19 (7) ◽

pp. 912-919 ◽

Cited By ~ 8

Author(s):

Meena Subramanyam ◽

Tatiana Plavina ◽

Bhupendra O Khatri ◽

Robert J Fox ◽

Susan E Goelz

Keyword(s):

Plasma Exchange ◽

Immune Reconstitution ◽

Jc Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Fold Reduction ◽

The Central Nervous System ◽

Igg Isotypes ◽

Circulating Antibodies ◽

Antibody Levels

Objective: Natalizumab, a highly effective treatment for multiple sclerosis (MS) and Crohn’s disease, is associated with progressive multifocal leukoencephalopathy (PML). Upon suspicion or diagnosis of PML, plasma exchange (PLEX) is performed to remove natalizumab from the circulation, allowing immune reconstitution of the central nervous system. Since PLEX may also remove other circulating antibodies, we examined the effects of PLEX on serum immunoglobulin (IgG) and anti–JC virus (JCV) antibody levels in MS patients with and without PML. Methods: Serum samples from 12 natalizumab-treated patients without PML collected before, during and after PLEX were tested for IgG isotypes using a commercial assay, and for anti-JCV antibodies using a two-step enzyme-linked immunosorbent assay. Five natalizumab-treated PML patients who underwent PLEX were also tested for anti-JCV antibodies. Results: PLEX produced a two- to three-fold reduction in all IgG isotypes. Among patients without PML, 42% (five of 12 patients) had detectable anti-JCV antibodies before PLEX; in these patients, anti-JCV antibodies were reduced approximately two- to five-fold, with levels returning to 50–100 percent of baseline two weeks after the final PLEX. The five PML patients, all of whom had detectable anti-JCV antibodies before PLEX, experienced similar reductions in anti-JCV antibody levels following PLEX. Conclusions: Our results indicate that PLEX effectively removes circulating antibodies; however, levels of endogenous anti-JCV antibody, unlike exogenously administered natalizumab, were replenished relatively quickly following PLEX. While interpretation of anti-JCV antibody levels during or within two weeks after PLEX may be problematic, humoral JCV immunity is not abolished by PLEX and antibody levels are rapidly restored.

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Significantly Low Levels of IgG Antibodies Against Oncogenic Merkel Cell Polyomavirus in Sera From Females Affected by Spontaneous Abortion

Frontiers in Microbiology ◽

10.3389/fmicb.2021.789991 ◽

2021 ◽

Vol 12 ◽

Author(s):

Chiara Mazziotta ◽

Giulia Pellielo ◽

Mauro Tognon ◽

Fernanda Martini ◽

John Charles Rotondo

Keyword(s):

Antibody Response ◽

Spontaneous Abortion ◽

Viral Infections ◽

Merkel Cell Polyomavirus ◽

Enzyme Linked Immunosorbent Assay ◽

Merkel Cell ◽

Igg Antibodies ◽

Igg Antibody ◽

The Impact

Merkel cell polyomavirus (MCPyV) is a small DNA tumor virus ubiquitous in humans. MCPyV establishes a clinically asymptomatic lifelong infection in healthy immunocompetent individuals. Viral infections are considered to be risk factors for spontaneous abortion (SA), which is the most common adverse complication of pregnancy. The role of MCPyV in SA remains undetermined. Herein, the impact of MCPyV infection in females affected by SA was investigated. Specifically, an indirect enzyme-linked immunosorbent assay (ELISA) method with two linear synthetic peptides/mimotopes mimicking MCPyV antigens was used to investigate immunoglobulin G (IgG) antibodies against MCPyV in sera from 94 females affected by SA [mean ± standard deviation (SD) age 35 ± (6) years] and from 96 healthy females undergoing voluntary pregnancy interruption [VI, mean (±SD) age 32 ± (7) years]. MCPyV seroprevalence and serological profiles were analyzed. The overall prevalence of serum IgG antibodies against MCPyV was 35.1% (33/94) and 37.5% (36/96) in SA and VI females, respectively (p > 0.05). Notably, serological profile analyses indicated lower optical densities (ODs) in females with SA compared to those undergoing VI (p < 0.05), thus indicating a reduced IgG antibody response in SA females. Circulating IgGs were identified in sera from SA and VI females. Our immunological findings indicate that a relatively reduced fraction of pregnant females carry serum anti-MCPyV IgG antibodies, while SA females presented a more pronounced decrease in IgG antibody response to MCPyV. Although yet to be determined, this immunological decrease might prompt an increase in MCPyV multiplication events in females experiencing abortive events. The role of MCPyV in SA, if present, remains to be determined.

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Horseradish Peroxidase-Conjugated-Nanobody-Based cELISA For The Rapid and Sensitive Clinical Detection of African Swine Fever Virus Antibodies

10.21203/rs.3.rs-1076003/v1 ◽

2021 ◽

Author(s):

Huijun Zhao ◽

Jiahui Ren ◽

Shuya Wu ◽

Yongkun Du ◽

Bo Wan ◽

...

Keyword(s):

Horseradish Peroxidase ◽

Fusion Protein ◽

Expression System ◽

African Swine Fever ◽

Enzyme Linked Immunosorbent Assay ◽

Hemorrhagic Disease ◽

Serum Samples ◽

Promising Alternative ◽

Technical Requirements ◽

Pig Serum

Abstract Background: African swine fever (ASF), which is caused by the ASF virus (ASFV), is a highly contagious hemorrhagic disease that affects pigs and has the potential to cause mortality in almost 100% of domestic pigs and wild boars. Due to the lack of an effective vaccine, the control of ASF must depend on early, efficient, cost-effective detection and strict control and elimination strategies. Traditional molecular and serological testing methods are generally associated with high testing costs, complex operations and high technical requirements. As a promising alternative diagnostic tool to traditional antibodies, nanobodies (Nb) have the advantages of simpler and faster generation, good stability and solubility, and high affinity and specificity. The application of Nbs in the detection of ASFV antibodies in the serum has not yet been reported, to the best of our knowledge. Results: Using a phage display technology, one specific Nb against the ASFV p54 protein that exhibited high specificity and affinity to the protein, Nb8, was successfully screened. A HEK293T cell line stably expressing Nb8-horseradish peroxidase (HRP) fusion protein was established using the lentiviral expression system. Following the optimization of the reaction conditions, the Nb8-HRP fusion protein was successfully used to establish a competitive enzyme-linked immunosorbent assay (cELISA) to detect ASFV-specific antibodies in pig serum, for the first time. The cut-off value for the cELISA was 15.78%. A total of 209 serum samples were tested using the developed cELISA and a commercial ELISA kit. The specificity of the cELISA was 98.97%, and the limit of detection was 1:320 in inactivated ASFV antibody-positive reference serum samples, with the coincidence rate between the two methods being 98.56%. Conclusions: A specific, sensitive and repeatable cELISA was successfully developed based on the unique Nb8 as a probe, providing a promising method for the detection of anti-ASFV antibodies in clinical pig serum.

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Serological evidence of vertical transmission of JC and BK polyomaviruses in humans

Journal of General Virology ◽

10.1099/vir.0.028571-0 ◽

2011 ◽

Vol 92 (5) ◽

pp. 1044-1050 ◽

Cited By ~ 34

Author(s):

Renzo Boldorini ◽

Sara Allegrini ◽

Umberto Miglio ◽

Alessia Paganotti ◽

Norma Cocca ◽

...

Keyword(s):

Pregnant Women ◽

Vertical Transmission ◽

Jc Virus ◽

Bk Virus ◽

Serological Evidence ◽

Serum Samples ◽

Virus Like Particle ◽

Genomic Study ◽

Nasopharyngeal Secretion ◽

Blood And Urine Samples

Vertical transmission of JC virus and BK virus has been investigated by few authors, with conflicting results. We performed a combined serological and genomic study of 19 unselected pregnant women and their newborns. Blood and urine samples were collected during each gestational trimester from the pregnant women. Umbilical cord blood, peripheral blood, urine and nasopharyngeal secretion samples were taken from newborns at delivery and after 1 week and 1 month of life. Polyomavirus DNA was detected by nested PCR. Polyomavirus IgG-, IgM- and IgA-specific antibodies were measured in maternal and newborn serum samples using a virus-like-particle-based ELISA method. BKV and JCV DNA were detected in urine from 4 (21 %) and 5 (26 %) women, respectively. BKV and JCV seroprevalences in the pregnant women were 84 % and 42 %, respectively. Using a rise in the IgG level or the transient appearance of an IgA or IgM response as evidence of infection in the newborn, we detected BKV and JCV infections in four (21 %) and three (16 %) newborns, respectively. Three infants had serological evidence of infection with both BKV and JCV. In two of the four possible BKV-infected newborns, the mothers seroconverted during pregnancy, while another mother was viruric and IgA seropositive. The mother of one of the three possible JCV-infected newborns was viruric and IgA seropositive; another mother was viruric. These results suggest JC virus and BK virus can be transmitted from mother to newborn during pregnancy or soon after birth.

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Optimization and Validation of Indirect ELISA Using Truncated TssB Protein for the Serodiagnosis of Glanders amongst Equines

The Scientific World JOURNAL ◽

10.1155/2014/469407 ◽

2014 ◽

Vol 2014 ◽

pp. 1-6 ◽

Cited By ~ 8

Author(s):

Harisankar Singha ◽

Praveen Malik ◽

Sachin K. Goyal ◽

Sandip K. Khurana ◽

Chiranjay Mukhopadhyay ◽

...

Keyword(s):

Indirect Elisa ◽

Expression System ◽

Cross Reactivity ◽

Amino Acid Sequences ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Diagnostic Efficacy ◽

Diagnostic Potential ◽

Prokaryotic Expression System ◽

Control Serum

Objective. To express truncated TssB protein ofBurkholderia malleiand to evaluate its diagnostic efficacy for serological detection of glanders among equines.Materials and Methods. In an attempt to develop recombinant protein based enzyme-linked immunosorbent assay (ELISA), N-terminal 200 amino acid sequences ofB. malleiTssB protein—a type 6 secretory effector protein—were expressed in prokaryotic expression system. Diagnostic potential of recombinant TssB protein was evaluated in indirect ELISA using a panel of glanders positive (n=49), negative (n=30), and field serum samples (n=1811). Cross-reactivity of the assay was assessed with equine disease control serum and human melioidosis positive serum.Results. In comparison to CFT, diagnostic sensitivity and specificity of ELISA were 99.7% and 100%, respectively.Conclusions. The indirect ELISA method using the truncated TssB offered safer and more rapid and efficient means of serodiagnosis of glanders in equines. These data highlight the use of TssB as potential diagnostic antigen for serological diagnosis of glanders.

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Use of MAG1 Recombinant Antigen for Diagnosis of Toxoplasma gondii Infection in Humans

Clinical and Vaccine Immunology ◽

10.1128/cvi.00419-06 ◽

2007 ◽

Vol 14 (3) ◽

pp. 220-225 ◽

Cited By ~ 32

Author(s):

Lucyna Holec ◽

Elżbieta Hiszczyńska-Sawicka ◽

Artur Gąsior ◽

Anna Brillowska-Dąbrowska ◽

Józef Kur

Keyword(s):

Toxoplasma Gondii ◽

Diagnostic Tests ◽

Expression System ◽

Recombinant Antigen ◽

Acute Stage ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Acute Toxoplasmosis ◽

Toxoplasma Gondii Infection ◽

Potential Use

ABSTRACT This paper describes the cloning, purification, and serological applications of matrix antigen MAG1 of Toxoplasma gondii. The expression system used allows the production of a large amount of T. gondii recombinant protein, which was assessed for its potential use in an enzyme-linked immunosorbent assay (ELISA) for detection of T. gondii infection in humans. Serum samples from 117 patients with different stages of infection, along with 10 serum samples from seronegative patients obtained for routine diagnostic tests, were used. The results were compared with those of an ELISA that uses a native T. gondii antigen extract. The MAG1 antigen detected antibodies more frequently from the acute stage (97.3%) than from the chronic stage (7.5%) of toxoplasmosis. Hence, this antigen may be used as a tool for detection of T. gondii immunoglobulin G antibodies in persons with acute toxoplasmosis.

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Prevalence and stability of human serum antibodies to simian virus 40 VP1 virus-like particles

Journal of General Virology ◽

10.1099/vir.0.80783-0 ◽

2005 ◽

Vol 86 (6) ◽

pp. 1703-1708 ◽

Cited By ~ 30

Author(s):

Annika Lundstig ◽

Linda Eliasson ◽

Matti Lehtinen ◽

Kestutis Sasnauskas ◽

Pentti Koskela ◽

...

Keyword(s):

Jc Virus ◽

Simian Virus 40 ◽

Simian Virus ◽

Bk Virus ◽

Confirmatory Test ◽

Human Populations ◽

Specific Antibodies ◽

Virus Like Particles ◽

Sv40 Dna ◽

Over Time

Possible human infection with simian virus 40 (SV40) has been of great concern ever since SV40 was discovered in polio vaccines. Human populations are SV40-seropositive, but because of serological cross-reactivity between SV40 and the human polyomaviruses BK virus (BKV) and JC virus (JCV), it is debatable whether these antibodies are specific. An SV40-specific serological assay was established, based on purified virus-like particles (VLPs), where the SV40 VLPs were blocked with hyperimmune sera to BKV and JCV. Competition with SV40 hyperimmune sera was used as a confirmatory test. Among 288 Swedish children of between 1 and 13 years of age, 7·6 % had SV40-specific antibodies. SV40 seroprevalence reached a peak of 14 % at 7–9 years of age. Among 100 control patients with benign tumours, 9 % were SV40-seropositive. However, SV40 DNA was not detectable in corresponding buffy-coat samples. In serial samples taken up to 5 years apart from 141 Finnish women participating in the population-based serological screening for congenital infections, only two of 141 women were SV40-seropositive in both samples. Six women seroconverted and eight women had a loss of antibodies over time. None of the SV40-seropositive samples contained detectable SV40 DNA. In conclusion, there is a low prevalence of SV40-specific antibodies in the Nordic population. The SV40 antibodies appear to have a low stability over time and their origin is not clear.

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