A capripoxvirus detection PCR and antibody ELISA based on the major antigen P32, the homolog of the vaccinia virus H3L gene

Heine, H.G.; Stevens, Matthew P.; Foord, Adam J.; Boyle, D.B

doi:10.1016/s0022-1759(99)00072-1

Cited by 136 publications

(87 citation statements)

References 12 publications

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“…(A separate group possibly studied the same protein but misidentified it as the VV IHD H6R gene product [25,26]). Similarly, antibodies to the orthologs of H3L are also seen after infection with other orthopoxviruses, including orf virus, capripox virus, and fowlpox virus (8,28,33). The H3L gene is strongly conserved among the orthopoxviruses.…”

Section: Discussionmentioning

confidence: 97%

Vaccinia Virus H3L Envelope Protein Is a Major Target of Neutralizing Antibodies in Humans and Elicits Protection against Lethal Challenge in Mice

Davies

McCausland

Valdez

et al. 2005

J Virol

201

216

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The smallpox vaccine is the prototypic vaccine, yet the viral targets critical for vaccine-mediated protection remain unclear in humans. We have produced protein microarrays of a near-complete vaccinia proteome and used them to determine the major antigen specificities of the human humoral immune response to the smallpox vaccine (Dryvax). H3L, an intracellular mature virion envelope protein, was consistently recognized by hightiter antibodies in the majority of human donors, particularly after secondary immunization. We then focused on examining H3L as a valuable human antibody target. Purified human anti-H3L antibodies exhibited substantial vaccinia virus-neutralizing activity in vitro (50% plaque reduction neutralization test [PRNT 50 ] ‫؍‬ 44 g/ml). Mice also make an immunodominant antibody response to H3L after vaccination with vaccinia virus, as determined by vaccinia virus protein microarray. Mice were immunized with recombinant H3L protein to examine H3L-specific antibody responses in greater detail. H3L-immunized mice developed hightiter vaccinia virus-neutralizing antibodies (mean PRNT 50 ‫؍‬ 1:3,760). Importantly, H3L-immunized mice were subsequently protected against lethal intranasal challenges with 1 or 5 50% lethal doses (LD 50 ) of pathogenic vaccinia virus strain WR, demonstrating the in vivo value of an anti-H3L response. To formally demonstrate that neutralizing anti-H3L antibodies are protective in vivo, we performed anti-H3L serum passive-transfer experiments. Mice receiving H3L-neutralizing antiserum were protected from a lethal challenge with 3 LD 50 of vaccinia virus strain WR (5/10 versus 0/10; P < 0.02). Together, these data show that H3L is a major target of the human anti-poxvirus antibody response and is likely to be a key contributor to protection against poxvirus infection and disease.

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Section: Discussionmentioning

confidence: 97%

Vaccinia Virus H3L Envelope Protein Is a Major Target of Neutralizing Antibodies in Humans and Elicits Protection against Lethal Challenge in Mice

Davies

McCausland

Valdez

et al. 2005

J Virol

201

216

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“…Indeed, the rational design of CaPV subspecies primers in this region may permit the development of conventional and real-time PCR allowing the direct differential detection of the three CaPVs. Such a tool would be more robust and less sensitive to genetic drift than the two conventional PCRs described before, which both rely on subsequent analysis of the restriction profiles of the PCR products (Heine et al, 1999;Hosamani et al, 2004). This tool would also supersede a recently developed conventional PCR for LSDV, which does not allow the detection of SPPV (Stram et al, 2008).…”

Section: Analysis Of Capripoxvirus Gpcr Genementioning

confidence: 99%

Capripoxvirus G-protein-coupled chemokine receptor: a host-range gene suitable for virus animal origin discrimination

Goff

Lamien

Fakhfakh

et al. 2009

Journal of General Virology

143

142

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The genus Capripoxvirus within the family Poxviridae comprises three closely related viruses, namely goat pox, sheep pox and lumpy skin disease viruses. This nomenclature is based on the animal species from which the virus was first isolated, respectively, goat, sheep and cattle. Since capripoxviruses are serologically identical, their specific identification relies exclusively on the use of molecular tools. We describe here the suitability of the G-protein-coupled chemokine receptor (GPCR) gene for use in host-range grouping of capripoxviruses. The analysis of 58 capripoxviruses showed three tight genetic clusters consisting of goat pox, sheep pox and lumpy skin disease viruses. However, a few discrepancies exist with the classical virus-host origin nomenclature: a virus isolated from sheep is grouped in the goat poxvirus clade and vice versa. Intra-group diversity was further observed for the goat pox and lumpy skin disease virus isolates. Despite the presence of nine vaccine strains, no genetic determinants of virulence were identified on the GPCR gene. For sheep poxviruses, the addition or deletion of 21 nucleic acids (7 aa) was consistently observed in the 59 terminal part of the gene. Specific signatures for each cluster were also identified. Prediction of the capripoxvirus GPCR topology, and its comparison with other known mammalian GPCRs and viral homologues, revealed not only a classical GPCR profile in the last three-quarters of the protein but also unique features such as a longer N-terminal end with a proximal hydrophobic a-helix and a shorter serine-rich C-tail.3These authors contributed equally to this work.Two supplementary tables are available with the online version of this paper.Journal of General Virology (2009), 90, 1967-1977 DOI 10.1099 Davies, 1981Davies, , 1982Diallo & Viljoen, 2007). CaPV infections lead to similar clinical signs in sheep and goats, mainly characterized by fever, excess salivation, conjunctivitis and rhinitis with ocular and nasal discharges. These symptoms are followed by eruption of pox lesions throughout the skin. LSD is a subacute to acute cattle disease with appearance of skin nodules. During epizootics of CaPVs in domestic animals, disease in wild ungulates has never been reported (Davies, 1991), although serology and isolated cases suggests that wildlife species have a possible role in virus maintenance.CaPVs are generally considered to be host-specific, leading to outbreaks in one preferred host. This is partially true since some SPPV and GTPV isolates are capable of causing severe diseases in both sheep and goats (Davies, 1976;Kitching et al., 1989). An in-depth epidemiological investigation to specifically identify the viruses involved in acute small ruminant pox diseases cannot be achieved by serological testing alone due to the very close antigenic relationship between CaPVs, with the existence of only one serotype (Kitching et al., , 1989. The common immunogenic properties of these viruses have been used for the preparation of live attenuated vaccines that ...

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“…This homology can be used for the developing of PCR based methods for the detection of capripoxviruses. On the other hand, the identifi ed sequence variability between LSDV, SPPV and GTPV occur in genes that likely determine virulence and host range [9] and were used for the developing of differential PCR protocols [10][11][12][13][14][15][16]. However, only a small number of tests are developed to distinguish strains within LSDV, especially the fi eld strain from the vaccine strain [17,18].…”

Section: Introductionmentioning

confidence: 99%

Real-Time PCR Assays for the Specific Detection of Field Balkan Strains of Lumpy Skin Disease Virus

Vidanović¹,

Šekler²,

Petrović

et al. 2016

Acta Veterinaria

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Lumpy skin di sease (LSD) is an important disease of cattle which is included in the OIE list of notifi able terrestrial animal diseases because of its great economic importance. The etiological agent is the Lumpy skin disease virus (LSDV).In the control of LSD attenuated strains of LSDV and SPPV are successfully used as vaccine strains in infected areas. In the case of vaccination policy, due to the possibility of mild or systemic post-vaccination reactions in vaccinated animals, the application of diagnostic procedures that will rapidly and specifi cally differentiate LSDV fi eld strains from LSD vaccine virus strains are extremely important. Rapidity in diagnostics and disposal of infected animals is one of the key factors in the prevention of spreading the disease.In the presented study we have described the development and validation of two realtime TaqMan-PCR assays for a rapid, sensitive and specifi c detection of the virulent fi eld LSDV strain currently circulating in the Balkan Peninsula. Specifi city for the fi eld strain and exclusivity for vaccine strains was tested on 171 samples from naturally infected and vaccinated animals.The results of this study show that both developed real-time PCR assays are more sensitive than the conventional nested PCR in detecting fi eld LSDV strains thus enabling rapid and high-throughput detection of animals infected with fi eld strains of LSDV.In conclusion, both KV-2 and FLI real-time PCR assays described in this study are simple, rapid, sensitive and suitable for routine use in a diagnostic laboratory and have the potential to replace conventional nested gel-based PCR assays as the standard procedure for the detection of fi eld strains of LSDV in clinical samples.

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A capripoxvirus detection PCR and antibody ELISA based on the major antigen P32, the homolog of the vaccinia virus H3L gene

Cited by 136 publications

References 12 publications

Vaccinia Virus H3L Envelope Protein Is a Major Target of Neutralizing Antibodies in Humans and Elicits Protection against Lethal Challenge in Mice

Vaccinia Virus H3L Envelope Protein Is a Major Target of Neutralizing Antibodies in Humans and Elicits Protection against Lethal Challenge in Mice

Capripoxvirus G-protein-coupled chemokine receptor: a host-range gene suitable for virus animal origin discrimination

Real-Time PCR Assays for the Specific Detection of Field Balkan Strains of Lumpy Skin Disease Virus

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