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Seroprevalence of Molluscum contagiosum virus in German and UK populations.

Sherwani S, Farleigh L, Agarwal N, Loveless S, Robertson N, Hadaschik E, Schnitzler P, Bugert JJ - PLoS ONE (2014)

Bottom Line: The overall seropositivity rate was found to be 14.8%.Ten out of 33 healthy UK individuals (30.3%; median age 27 years) had detectable MC084 antibodies.MCV seroconversion was more common in dermatological and autoimmune disorders, than in immunocompromised patients or in patients with multiple sclerosis.

View Article: PubMed Central - PubMed

Affiliation: Cardiff University School of Medicine, Institute of Infection and Immunity/Medical Microbiology, Cardiff, United Kingdom.

ABSTRACT
Molluscum contagiosum virus (MCV) is a significant but underreported skin pathogen for children and adults. Seroprevalence studies can help establish burden of disease. Enzyme linked immunosorbent assay (ELISA) based studies have been published for Australian and Japanese populations and the results indicate seroprevalences between 6 and 22 percent in healthy individuals, respectively. To investigate seroprevalence in Europe, we have developed a recombinant ELISA using a truncated MCV virion surface protein MC084 (V123-R230) expressed in E. coli. The ELISA was found to be sensitive and specific, with low inter- and intra-assay variability. Sera from 289 German adults and children aged 0-40 years (median age 21 years) were analysed for antibodies against MC084 by direct binding ELISA. The overall seropositivity rate was found to be 14.8%. The seropositivity rate was low in children below the age of one (4.5%), peaked in children aged 2-10 years (25%), and fell again in older populations (11-40 years; 12.5%). Ten out of 33 healthy UK individuals (30.3%; median age 27 years) had detectable MC084 antibodies. MCV seroconversion was more common in dermatological and autoimmune disorders, than in immunocompromised patients or in patients with multiple sclerosis. Overall MCV seroprevalence is 2.1 fold higher in females than in males in a UK serum collection. German seroprevalences determined in the MC084 ELISA (14.8%) are at least three times higher than incidence of MC in a comparable Swiss population (4.9%). While results are not strictly comparable, this is lower than Australian seroprevalence in a virion based ELISA (n = 357; 23%; 1999), but higher than the seroprevalence reported in a Japanese study using an N-terminal truncation of MC133 (n = 108, 6%; 2000. We report the first large scale serological survey of MC in Europe (n = 393) and the first MCV ELISA based on viral antigen expressed in E. coli.

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Bioinformatics.(A) Transmembrane plot (TMHMM Server v.2.0) [25] of mc084 amino acids 1–318; (B) hydropathy plot of MC084 protein with predicted high hydrophilic/antigenic regions indicated by black boxes. The full length ORF (MC084 1–318; predicted molecular weight 34.2 kD; shown on top) was cloned into vRB12 using specific primers tailed with restriction enzyme sites BamHI-HindIII) and C-terminal StrepII epitope tag. The resulting plasmid p319 was sequenced and the recombinant vaccinia virus v319 isolated on BSC-1 cells using the plaqueless mutant system [26]. N- and C-terminal (in yellow) truncations were subcloned from the original full length MCV gene into pGEX-2TK for overexpression in E. coli BL21 (RIL+). TMHMM was used to determine transmembrane regions [26] whereas the Kyte-Doolittle plot was used to identify hydrophilic regions with predicted high antigenicity [27].
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pone-0088734-g001: Bioinformatics.(A) Transmembrane plot (TMHMM Server v.2.0) [25] of mc084 amino acids 1–318; (B) hydropathy plot of MC084 protein with predicted high hydrophilic/antigenic regions indicated by black boxes. The full length ORF (MC084 1–318; predicted molecular weight 34.2 kD; shown on top) was cloned into vRB12 using specific primers tailed with restriction enzyme sites BamHI-HindIII) and C-terminal StrepII epitope tag. The resulting plasmid p319 was sequenced and the recombinant vaccinia virus v319 isolated on BSC-1 cells using the plaqueless mutant system [26]. N- and C-terminal (in yellow) truncations were subcloned from the original full length MCV gene into pGEX-2TK for overexpression in E. coli BL21 (RIL+). TMHMM was used to determine transmembrane regions [26] whereas the Kyte-Doolittle plot was used to identify hydrophilic regions with predicted high antigenicity [27].

Mentions: Amino acid sequences of MC084 (298 aa, 34 kD) were analysed to determine overall homology with related proteins in the GenBank and identify transmembrane regions and region of high hydrophilicity/high antigenicity. Two transmembrane regions predicted in the C-terminal end of the protein [26], were excluded to avoid solubility issues in the E. coli expression system (Figure 1A). Of the remaining amino acids, a N-terminal region (V33-G117) and a C-terminal region (aa V123-R230), both containing one region of high hydrophilicity in the Kyte–Doolittle plot (Figure 1B) [28] were further analysed for subcloning.


Seroprevalence of Molluscum contagiosum virus in German and UK populations.

Sherwani S, Farleigh L, Agarwal N, Loveless S, Robertson N, Hadaschik E, Schnitzler P, Bugert JJ - PLoS ONE (2014)

Bioinformatics.(A) Transmembrane plot (TMHMM Server v.2.0) [25] of mc084 amino acids 1–318; (B) hydropathy plot of MC084 protein with predicted high hydrophilic/antigenic regions indicated by black boxes. The full length ORF (MC084 1–318; predicted molecular weight 34.2 kD; shown on top) was cloned into vRB12 using specific primers tailed with restriction enzyme sites BamHI-HindIII) and C-terminal StrepII epitope tag. The resulting plasmid p319 was sequenced and the recombinant vaccinia virus v319 isolated on BSC-1 cells using the plaqueless mutant system [26]. N- and C-terminal (in yellow) truncations were subcloned from the original full length MCV gene into pGEX-2TK for overexpression in E. coli BL21 (RIL+). TMHMM was used to determine transmembrane regions [26] whereas the Kyte-Doolittle plot was used to identify hydrophilic regions with predicted high antigenicity [27].
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3928281&req=5

pone-0088734-g001: Bioinformatics.(A) Transmembrane plot (TMHMM Server v.2.0) [25] of mc084 amino acids 1–318; (B) hydropathy plot of MC084 protein with predicted high hydrophilic/antigenic regions indicated by black boxes. The full length ORF (MC084 1–318; predicted molecular weight 34.2 kD; shown on top) was cloned into vRB12 using specific primers tailed with restriction enzyme sites BamHI-HindIII) and C-terminal StrepII epitope tag. The resulting plasmid p319 was sequenced and the recombinant vaccinia virus v319 isolated on BSC-1 cells using the plaqueless mutant system [26]. N- and C-terminal (in yellow) truncations were subcloned from the original full length MCV gene into pGEX-2TK for overexpression in E. coli BL21 (RIL+). TMHMM was used to determine transmembrane regions [26] whereas the Kyte-Doolittle plot was used to identify hydrophilic regions with predicted high antigenicity [27].
Mentions: Amino acid sequences of MC084 (298 aa, 34 kD) were analysed to determine overall homology with related proteins in the GenBank and identify transmembrane regions and region of high hydrophilicity/high antigenicity. Two transmembrane regions predicted in the C-terminal end of the protein [26], were excluded to avoid solubility issues in the E. coli expression system (Figure 1A). Of the remaining amino acids, a N-terminal region (V33-G117) and a C-terminal region (aa V123-R230), both containing one region of high hydrophilicity in the Kyte–Doolittle plot (Figure 1B) [28] were further analysed for subcloning.

Bottom Line: The overall seropositivity rate was found to be 14.8%.Ten out of 33 healthy UK individuals (30.3%; median age 27 years) had detectable MC084 antibodies.MCV seroconversion was more common in dermatological and autoimmune disorders, than in immunocompromised patients or in patients with multiple sclerosis.

View Article: PubMed Central - PubMed

Affiliation: Cardiff University School of Medicine, Institute of Infection and Immunity/Medical Microbiology, Cardiff, United Kingdom.

ABSTRACT
Molluscum contagiosum virus (MCV) is a significant but underreported skin pathogen for children and adults. Seroprevalence studies can help establish burden of disease. Enzyme linked immunosorbent assay (ELISA) based studies have been published for Australian and Japanese populations and the results indicate seroprevalences between 6 and 22 percent in healthy individuals, respectively. To investigate seroprevalence in Europe, we have developed a recombinant ELISA using a truncated MCV virion surface protein MC084 (V123-R230) expressed in E. coli. The ELISA was found to be sensitive and specific, with low inter- and intra-assay variability. Sera from 289 German adults and children aged 0-40 years (median age 21 years) were analysed for antibodies against MC084 by direct binding ELISA. The overall seropositivity rate was found to be 14.8%. The seropositivity rate was low in children below the age of one (4.5%), peaked in children aged 2-10 years (25%), and fell again in older populations (11-40 years; 12.5%). Ten out of 33 healthy UK individuals (30.3%; median age 27 years) had detectable MC084 antibodies. MCV seroconversion was more common in dermatological and autoimmune disorders, than in immunocompromised patients or in patients with multiple sclerosis. Overall MCV seroprevalence is 2.1 fold higher in females than in males in a UK serum collection. German seroprevalences determined in the MC084 ELISA (14.8%) are at least three times higher than incidence of MC in a comparable Swiss population (4.9%). While results are not strictly comparable, this is lower than Australian seroprevalence in a virion based ELISA (n = 357; 23%; 1999), but higher than the seroprevalence reported in a Japanese study using an N-terminal truncation of MC133 (n = 108, 6%; 2000. We report the first large scale serological survey of MC in Europe (n = 393) and the first MCV ELISA based on viral antigen expressed in E. coli.

Show MeSH
Related in: MedlinePlus