Detection of African horsesickness virus in infected spleens by a sandwich ELISA using two monoclonal antibodies specific for VP7

Laviada, M. D.; Babín, Mar; Dominguez, J.; Sánchez-Vizcaíno, José Manuel

doi:10.1016/0166-0934(92)90113-r

Cited by 35 publications

(20 citation statements)

References 17 publications

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“…The indirect sandwich ELISA is also extremely useful for the rapid identification of AHSV antigen in solid tissues taken from animals that have died following an acute infection [33,45,67]. In addition, isolated viruses can be identified by group specific tests such as complement fixation [75] and direct and indirect fluorescence [27].…”

Section: Antigen Identificationmentioning

confidence: 99%

African horse sickness

2004

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-African horse sickness virus (AHSV) causes a non-contagious, infectious insect-borne disease of equids and is endemic in many areas of sub-Saharan Africa and possibly Yemen in the Arabian Peninsula. However, periodically the virus makes excursions beyond its endemic areas and has at times extended as far as India and Pakistan in the east and Spain and Portugal in the west. The vectors are certain species of Culicoides biting midge the most important of which is the AfroAsiatic species C. imicola. This paper describes the effects that AHSV has on its equid hosts, aspects of its epidemiology, and present and future prospects for control. The distribution of AHSV seems to be governed by a number of factors including the efficiency of control measures, the presence or absence of a long term vertebrate reservoir and, most importantly, the prevalence and seasonal incidence of the major vector which is controlled by climate. However, with the advent of climatechange the major vector, C. imicola, has now significantly extended its range northwards to include much of Portugal, Spain, Italy and Greece and has even been recorded from southern Switzerland. Furthermore, in many of these new locations the insect is present and active throughout the entire year. With the related bluetongue virus, which utilises the same vector species of Culicoides this has, since 1998, precipitated the worst outbreaks of bluetongue disease ever recorded with the virus extending further north in Europe than ever before and apparently becoming endemic in that continent. The prospects for similar changes in the epidemiology and distribution of AHSV are discussed.

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Section: Antigen Identificationmentioning

confidence: 99%

African horse sickness

2004

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“…Viraemia titres of each viraemic blood sample were determined by plaque assay on Vero cell monolayers as described previously (Oellermann, 1970). CPE was confirmed by an AHSV-specific DAS-ELISA (Laviada et al, 1992b).…”

Section: Virus Neutralization Test and Viraemia Assaysmentioning

confidence: 99%

Full protection against African horsesickness (AHS) in horses induced by baculovirus-derived AHS virus serotype 4 VP2, VP5 and VP7

Diaz-Laviada

Roy

et al. 1996

Journal of General Virology

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African horsesickness virus serotype 4 (AHSV-4) outer capsid protein VP2, or VP2 and VP5 plus inner capsid protein VP7, derived from single or dual recombinant baculovirus expression vectors were used in different combinations to immunize horses. When the proteins were purified by affinity chromatography, the combination of all three proteins induced low levels of neutralizing antibodies and conferred protection against virulent virus challenge. However, purified VP2 or VP2 and VP5 in the absence of VP7 failed to induce neutralizing antibodies and protection. Immunization with non-purified proteins enhanced the titres of neutralizing antibodies. Again, the combination of the three proteins was able to confer total protection to immunized horses, which showed absence of viraemia. The antigenicity of recombinant VP2 was analysed with a collection of 30 MAbs. Both purified and unpurified recombinant VP2 proteins showed different antigenic patterns in comparison to that of VP2 on virions. An immunization experiment with four more horses confirmed these results. The vaccine described here would not only prevent the disease, but would drastically reduce the propagation of the virus by vectors.

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“…When the first signs of c.p.e, were observed (at 2 days p.i. ), cells were labelled with 500 pCi [aSS]methionine in 1 ml of medium without methionine and 2 % foetal calf serum for 24 h. Cells were harvested, mixed with unlabelled infected cells from 20 roller bottles and purified as described (Laviada et al, 1992).…”

Section: Methodsmentioning

confidence: 99%

“…A Spanish strain of AHSV isolated in 1989 from an infected horse was grown in Vero cells as described (Laviada et al, 1992) and used after three to seven passages.…”

Section: Methodsmentioning

confidence: 99%

Characterization of African horsesickness virus serotype 4-induced polypeptides in Vero cells and their reactivity in Western immunoblotting

Laviada¹,

Arias²,

Sánchez-Vizcaíno³

1993

Journal of General Virology

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The structural and non-structural proteins induced by African horsesickness virus serotype 4 (AHSV-4) in infected Vero cells were analysed by SDS-PAGE. Twenty-two virus-induced polypeptides were detected in infected cells by comparison with the polypeptides of mock-infected cells, of which four major (VP2, VP3, VP5 and VP7) and three minor (VP1, VP4 and VP6) structural proteins and four non-structural proteins (P58, P48, P21 and P20) were shown to be virus-coded, as deduced from electrophoretic and antigenic studies of purified virions and infected cells. The proteins that elicit the major antibody responses both in vaccinated and naturally or experimentally infected horses were shown to be three structural proteins, VP2, VP5 and VP7, and the four major non-structural proteins, P58, P48, P21 and P20, as deduced by radioimmunoprecipitation and immunoblotting assays. The crossreactivity between AHSV-4 and sera obtained from horses experimentally infected with seven other serotypes was also determined. The results showed that VP5, VP7, P48, P21 and P20 are conserved and can be used to diagnose the infection of any of these eight serotypes.

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Detection of African horsesickness virus in infected spleens by a sandwich ELISA using two monoclonal antibodies specific for VP7

Cited by 35 publications

References 17 publications

African horse sickness

African horse sickness

Full protection against African horsesickness (AHS) in horses induced by baculovirus-derived AHS virus serotype 4 VP2, VP5 and VP7

Characterization of African horsesickness virus serotype 4-induced polypeptides in Vero cells and their reactivity in Western immunoblotting

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