Limits...
Serologic evidence for novel poxvirus in endangered red Colobus monkeys, Western Uganda.

Goldberg TL, Chapman CA, Cameron K, Saj T, Karesh WB, Wolfe ND, Wong SW, Dubois ME, Slifka MK - Emerging Infect. Dis. (2008)

Bottom Line: Enzyme-linked immunosorbent assay, Western blot, and virus neutralization assays indicated that red colobus monkeys in Kibale National Park, western Uganda, had antibodies to a virus that was similar, but not identical, to known orthopoxviruses.The presence of a novel poxvirus in this endangered primate raises public health and conservation concerns.

View Article: PubMed Central - PubMed

Affiliation: University of Illinois, College of Veterinary Medicine, Department of Pathobiology, 2001 South Lincoln Ave, Urbana, IL 61802, USA. tlgoldbe@uiuc.edu

ABSTRACT
Enzyme-linked immunosorbent assay, Western blot, and virus neutralization assays indicated that red colobus monkeys in Kibale National Park, western Uganda, had antibodies to a virus that was similar, but not identical, to known orthopoxviruses. The presence of a novel poxvirus in this endangered primate raises public health and conservation concerns.

Show MeSH

Related in: MedlinePlus

Western blot analysis of Orthopoxvirus (OPV)–reactive antibody responses in red colobus. Western blot analysis was performed to further characterize humoral immune responses against OPV antigens. Purified monkeypox virus (MPV), vaccinia virus (VV), and cowpox virus (CPV) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to polyvinylidene difluoride membranes, and probed with plasma from a VV-immune human, MPV-immune human, MVA-immune RM, MPV-immune RM, and 5 representative red colobus. The red colobus animal identification number is shown in the upper left corner of each Western blot for comparison with the ELISA data for the same sample described in Figure 1.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2600227&req=5

Figure 2: Western blot analysis of Orthopoxvirus (OPV)–reactive antibody responses in red colobus. Western blot analysis was performed to further characterize humoral immune responses against OPV antigens. Purified monkeypox virus (MPV), vaccinia virus (VV), and cowpox virus (CPV) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to polyvinylidene difluoride membranes, and probed with plasma from a VV-immune human, MPV-immune human, MVA-immune RM, MPV-immune RM, and 5 representative red colobus. The red colobus animal identification number is shown in the upper left corner of each Western blot for comparison with the ELISA data for the same sample described in Figure 1.

Mentions: Western blot analysis was performed to characterize more fully the antipoxvirus response of the red colobus. Two micrograms of purified MPV, VV, and CPV viral proteins were separated by 4%–20% gradient sodium dodecyl sulfate–polyacrylamide gel electrophoresis, transferred to polyvinylidene difluoride membranes (Invitrogen, Carlsbad, CA, USA), and probed with plasma (1:10,000) before addition of HRP-conjugated polyclonal goat antihuman Ig G (γ-specific, Jackson Immunolabs, Inc., West Grove, PA, USA) and chemiluminescent detection (Pierce SuperSignal West Dura Substrate, Rockville, IL, USA) (Figure 2). VV-immune human, MPV-immune human, MVA (modified vaccinia Ankara)-immune RM and MPV-immune RM plasma samples were included as positive controls to identify banding patterns typically observed in orthopoxvirus-immune humans and nonhuman primates. Western blot analysis proved more sensitive than anti-VV ELISA (Appendix Figure), with plasma from 30 of 31 red colobus reacting with at least 1 protein band from MPV, VV, or CPV. However, unlike the orthopoxvirus-immune human and RM controls, samples from red colobus demonstrated fewer immunoreactive bands and different immunodominant banding patterns, suggesting infection with either a distantly related orthopoxvirus or a virus from a different genus in the Poxviridae family.


Serologic evidence for novel poxvirus in endangered red Colobus monkeys, Western Uganda.

Goldberg TL, Chapman CA, Cameron K, Saj T, Karesh WB, Wolfe ND, Wong SW, Dubois ME, Slifka MK - Emerging Infect. Dis. (2008)

Western blot analysis of Orthopoxvirus (OPV)–reactive antibody responses in red colobus. Western blot analysis was performed to further characterize humoral immune responses against OPV antigens. Purified monkeypox virus (MPV), vaccinia virus (VV), and cowpox virus (CPV) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to polyvinylidene difluoride membranes, and probed with plasma from a VV-immune human, MPV-immune human, MVA-immune RM, MPV-immune RM, and 5 representative red colobus. The red colobus animal identification number is shown in the upper left corner of each Western blot for comparison with the ELISA data for the same sample described in Figure 1.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2600227&req=5

Figure 2: Western blot analysis of Orthopoxvirus (OPV)–reactive antibody responses in red colobus. Western blot analysis was performed to further characterize humoral immune responses against OPV antigens. Purified monkeypox virus (MPV), vaccinia virus (VV), and cowpox virus (CPV) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to polyvinylidene difluoride membranes, and probed with plasma from a VV-immune human, MPV-immune human, MVA-immune RM, MPV-immune RM, and 5 representative red colobus. The red colobus animal identification number is shown in the upper left corner of each Western blot for comparison with the ELISA data for the same sample described in Figure 1.
Mentions: Western blot analysis was performed to characterize more fully the antipoxvirus response of the red colobus. Two micrograms of purified MPV, VV, and CPV viral proteins were separated by 4%–20% gradient sodium dodecyl sulfate–polyacrylamide gel electrophoresis, transferred to polyvinylidene difluoride membranes (Invitrogen, Carlsbad, CA, USA), and probed with plasma (1:10,000) before addition of HRP-conjugated polyclonal goat antihuman Ig G (γ-specific, Jackson Immunolabs, Inc., West Grove, PA, USA) and chemiluminescent detection (Pierce SuperSignal West Dura Substrate, Rockville, IL, USA) (Figure 2). VV-immune human, MPV-immune human, MVA (modified vaccinia Ankara)-immune RM and MPV-immune RM plasma samples were included as positive controls to identify banding patterns typically observed in orthopoxvirus-immune humans and nonhuman primates. Western blot analysis proved more sensitive than anti-VV ELISA (Appendix Figure), with plasma from 30 of 31 red colobus reacting with at least 1 protein band from MPV, VV, or CPV. However, unlike the orthopoxvirus-immune human and RM controls, samples from red colobus demonstrated fewer immunoreactive bands and different immunodominant banding patterns, suggesting infection with either a distantly related orthopoxvirus or a virus from a different genus in the Poxviridae family.

Bottom Line: Enzyme-linked immunosorbent assay, Western blot, and virus neutralization assays indicated that red colobus monkeys in Kibale National Park, western Uganda, had antibodies to a virus that was similar, but not identical, to known orthopoxviruses.The presence of a novel poxvirus in this endangered primate raises public health and conservation concerns.

View Article: PubMed Central - PubMed

Affiliation: University of Illinois, College of Veterinary Medicine, Department of Pathobiology, 2001 South Lincoln Ave, Urbana, IL 61802, USA. tlgoldbe@uiuc.edu

ABSTRACT
Enzyme-linked immunosorbent assay, Western blot, and virus neutralization assays indicated that red colobus monkeys in Kibale National Park, western Uganda, had antibodies to a virus that was similar, but not identical, to known orthopoxviruses. The presence of a novel poxvirus in this endangered primate raises public health and conservation concerns.

Show MeSH
Related in: MedlinePlus