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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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Protein purification.Characterisation of over expressed recombinant fusion protein GST-MC084S and FPLC purified recombinant MC084S protein by SDS-PAGE and Western Blot. M: Molecular weight markers expressed in kDa. (A) Over-expressed 40 kDa Recombinant GST-MC084S fusion protein separated in a 4–12% Bis-Tris gel. (B) FPLC purified 14 kDa protein separated in a 15% Bis-Arylamide gel. Both gels were stained with Coomassie Brilliant Blue R-250. (C) GST-MC084S fusion protein after transfer to nitrocellulose (D) FPLC purified MC084S. The membranes were probed with Strep MAB-Classic HRP conjugate (IBA-lifesciences). Arrow heads indicate the locations of proteins.
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pone-0088734-g003: Protein purification.Characterisation of over expressed recombinant fusion protein GST-MC084S and FPLC purified recombinant MC084S protein by SDS-PAGE and Western Blot. M: Molecular weight markers expressed in kDa. (A) Over-expressed 40 kDa Recombinant GST-MC084S fusion protein separated in a 4–12% Bis-Tris gel. (B) FPLC purified 14 kDa protein separated in a 15% Bis-Arylamide gel. Both gels were stained with Coomassie Brilliant Blue R-250. (C) GST-MC084S fusion protein after transfer to nitrocellulose (D) FPLC purified MC084S. The membranes were probed with Strep MAB-Classic HRP conjugate (IBA-lifesciences). Arrow heads indicate the locations of proteins.

Mentions: The GST fusion protein was identified with an apparent molecular weight (MW) similar to the predicted MW of 40 kD in IPTG induced cultures (Figure 3A) and was absent in the uninduced cultures. The protein was protease cleaved and the C-terminal truncation of MC084 with an apparent MW of 14 kD was further purified via FPLC using a size exclusion Superdex S200 column (Figure 3B). The Strep II tag was identified in Western Blot in both the fusion protein and the cleaved MC084 (V123-R230)-Strep II truncation (Figures 3 C and D). Additional data on antigen selection and optimization can be found in Figures S1 and S2.


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)

Protein purification.Characterisation of over expressed recombinant fusion protein GST-MC084S and FPLC purified recombinant MC084S protein by SDS-PAGE and Western Blot. M: Molecular weight markers expressed in kDa. (A) Over-expressed 40 kDa Recombinant GST-MC084S fusion protein separated in a 4–12% Bis-Tris gel. (B) FPLC purified 14 kDa protein separated in a 15% Bis-Arylamide gel. Both gels were stained with Coomassie Brilliant Blue R-250. (C) GST-MC084S fusion protein after transfer to nitrocellulose (D) FPLC purified MC084S. The membranes were probed with Strep MAB-Classic HRP conjugate (IBA-lifesciences). Arrow heads indicate the locations of proteins.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0088734-g003: Protein purification.Characterisation of over expressed recombinant fusion protein GST-MC084S and FPLC purified recombinant MC084S protein by SDS-PAGE and Western Blot. M: Molecular weight markers expressed in kDa. (A) Over-expressed 40 kDa Recombinant GST-MC084S fusion protein separated in a 4–12% Bis-Tris gel. (B) FPLC purified 14 kDa protein separated in a 15% Bis-Arylamide gel. Both gels were stained with Coomassie Brilliant Blue R-250. (C) GST-MC084S fusion protein after transfer to nitrocellulose (D) FPLC purified MC084S. The membranes were probed with Strep MAB-Classic HRP conjugate (IBA-lifesciences). Arrow heads indicate the locations of proteins.
Mentions: The GST fusion protein was identified with an apparent molecular weight (MW) similar to the predicted MW of 40 kD in IPTG induced cultures (Figure 3A) and was absent in the uninduced cultures. The protein was protease cleaved and the C-terminal truncation of MC084 with an apparent MW of 14 kD was further purified via FPLC using a size exclusion Superdex S200 column (Figure 3B). The Strep II tag was identified in Western Blot in both the fusion protein and the cleaved MC084 (V123-R230)-Strep II truncation (Figures 3 C and D). Additional data on antigen selection and optimization can be found in Figures S1 and S2.

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