Solid-phase radioimmunoassay techniques for the detection of African swine fever antigen and antibody

Crowther, John R.; Wardley, R.C.; Wilkinson, Philip

doi:10.1017/s0022172400026140

Cited by 11 publications

(5 citation statements)

References 9 publications

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“…In endemic areas, diagnosis may be based on detection of antibody but in new introductions of disease it is preferable to detect the virus. Procedures, which have been used for ASF virus antigen detection, include the traditional haemadsorption test (HAD; [ 9 ], radioimmunoassay (RIA; [ 10 ];, direct immunofluorescence [ 11 ] and polymerase chain reaction (PCR; [ 12 - 14 ] and recently LAMP [ 15 ]. Immunocytochemistry and in situ hybridization have been described in studies of disease pathogenesis [ 16 ], but these techniques are not ideally suited to routine diagnosis [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

Prevalence of African swine fever virus in apparently healthy domestic pigs in Uganda

et al. 2013

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BackgroundAfrican swine fever (ASF) is a contagious viral disease which can cause up to 100% mortality among domestic pigs leading to serious socio-economic impact on people’s livelihoods. ASF is endemic in Uganda and there is paucity of information on the epidemiology of the disease. The major aim of this study was to determine the seroprevalence and prevalence of African swine fever virus (ASFV) in apparently healthy slaughter pigs at Wambizi slaughterhouse in Kampala city, Uganda. We also estimated the presence of ASFV antibodies and circulating viral antigens in pigs from selected districts of Uganda during targeted surveillance. We analysed 540 and 181 blood samples collected from slaughter pigs and pigs from targeted surveillance districts respectively.ResultsThe prevalence of ASFV in slaughter pigs was 52.96% (95% CI, 48.75-57.14) and 11.5% (95% CI, 9.06-14.45) by ELISA and PCR respectively. In surveillance districts, the proportion of ASFV positive pigs was 53.59% (95% CI, 46.33-60.71) and 0.55% (95% CI, 0.1-3.06) by ELISA and PCR respectively.ConclusionThe study has found out a high seroprevalence of ASFV antibodies in apparently healthy slaughter pigs and also a high proportion of ASFV antibody seropositive pigs in surveyed districts in Uganda indicating exposure to ASFV. However, there was a lower prevalence of ASFV infection implying that there could be low virulent strains of ASFV circulating in domestic pigs in Uganda which requires further investigation.

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Section: Introductionmentioning

confidence: 99%

Prevalence of African swine fever virus in apparently healthy domestic pigs in Uganda

et al. 2013

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“…Viruses could be grouped according to their ability to compete with the homologous virus, and statistically significant differences between virus “groups” were observed. The results of a comparison of the relationships between the viruses using RIA and complement fixation tests did not always correlate (Crowther et al 1979 ). RNA competition hybridization using 32 P-labeled RNA and competitive radioimmunoassays with 35 S-labeled viruses were used to compare the biochemical and serological characteristics of five isolate of FMD, serotype A (Robson et al 1979 ).…”

Section: Introductionmentioning

confidence: 99%

“…Enzyme immunoassays, immunoblotting, and indirect fluorescent antibody tests have all been used in diagnosis. Solid phase RIA using microtiter plates and 125 I-labeled IgG has been used to detect both viral antigen and antibody (Crowther et al 1979 ) and was shown to be 1,000 times more sensitive than an immune-electro-osmophoresis test for detection of ASFV antibody. RIA detected antibodies within 3–4 days of infection, in contrast with the gel diffusion tests that was not positive until 10 days post-infection (Wardley and Wilkinson 1980 ).…”

Section: Introductionmentioning

confidence: 99%

The role of nuclear technologies in the diagnosis and control of livestock diseases—a review

Viljoen

Luckins

2012

Trop Anim Health Prod

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Nuclear and nuclear-related technologies have played an important role in animal health, particularly in relation to disease diagnosis and characterization of pathogenic organisms. This review focuses primarily on how and where nuclear technologies, both non-isotopic and isotopic methods, have made their impact in the past and where it might be expected they could have an impact in the future. The review outlines the extensive use of radiation attenuation in attempts to create vaccines for a multiplicity of pathogenic organisms and how the technology is being re-examined in the light of recent advances in irradiation techniques and cryopreservation/lyophilization that might obviate some of the problems of maintenance of viable, attenuate vaccines and their transport and use in the field. This approach could be used for a number of parasitic diseases where vaccination has been problematic and where investigations into the development of molecular vaccines have still failed to deliver satisfactory candidates for generating protective immune responses. Irradiation of antigens or serum samples also has its uses in diagnosis, especially when the samples need to be transported across international boundaries, or when handling the pathogens in question when carrying out a test presents serious health hazards to laboratory personnel. The present-day extensive use of enzyme immunoassays and molecular methods (e.g., polymerase chain reaction) for diagnosis and characterization of animal pathogens has its origins in the use of isotope-labeled antigens and antibodies. These isotopic techniques that included the use of 75Se, 32P, 125I, and 35S isotopes enabled a level of sensitivity and specificity that was hitherto unrealized, and it is prescient to remind ourselves of just how successful these technologies were, in spite of their infrequent use nowadays. Finally, the review looks at the potential for stable isotope analysis for a variety of applications—in the tracking of animal migrations, where the migrant are potential carriers of transboundary animal diseases, and where it would be useful to determine the origins of the carrier, e.g., Highly Pathogenic Avian Influenza and its dissemination by wild water fowl. Other applications could be in monitoring sequestered microbial culture (e.g., rinderpest virus) where in the case of accidental or deliberate release of infective culture it would be possible to identify the laboratory from which the isolate originated.

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“…Swine infected with ASFV develop anti-viral antibodies that can be demonstrated from 7 days post-infection (p.i.) by a wide variety of tests (Cowan, 1961 ; Crowther et al , 1979 ; Hamdy et al , 1981 ; Malmquist, 1963 ; Pan et al , 1972 , 1974 , 1982 ; Parker & Plowright, 1968 ). Pigs that recover are usually resistant to challenge with homologous, but not heterologous, virus isolates (Ruiz-Gonzalvo et al , 1986 ; Schlafer et al , 1984 ).…”

Section: Introductionmentioning

confidence: 99%

Identification of the principal serological immunodeterminants of African swine fever virus by screening a virus cDNA library with antibody

Kollnberger

Gutiérrez-Castañeda

Foster-Cuevas

et al. 2002

Journal of General Virology

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Protective immunity to African swine fever virus (ASFV) may involve a combination of both serological and cellular mechanisms. This work is focused on the identification of the possible relevant serological immunodeterminants of immunity. Thus, 14 serological immunodeterminants of ASFV have been characterized by exhaustive screening of a representative lambda phage cDNA expression library of the tissue culture-adapted Ba71V strain of ASFV. The library was constructed using RNA extracted from Vero cells infected for 3, 6, 9 and 12 h. A total of 150 clones was selected arbitrarily by antibody screening of the library with a polyclonal antiserum from a domestic pig surviving infection with the virulent Malta isolate of ASFV. Sequencing of these clones permitted identification of 14 independent viral proteins that stimulated an antibody response. These included six proteins encoded by previously unassigned open reading frames (ORFs) (B602L, C44L, CP312R, E184L, K145R and K205R) as well as some of the more well-studied structural (A104R, p10, p32, p54 and p73) and non-structural proteins (RNA reductase, DNA ligase and thymidine kinase). Immunogenicity of these proteins was confirmed by demonstrating the corresponding antibodies in sera from pigs infected either with the Malta isolate or with the OURT88/3-OURT88/1 isolate combination. Furthermore, the majority of these ORFs were also recognized by immune antiserum from the natural host, the bush pig, following secondary challenge with the virulent Malawi (SINT90/1) isolate of ASFV. Thus, it is possible that some of these determinants may be important in protection against virus infection.

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Solid-phase radioimmunoassay techniques for the detection of African swine fever antigen and antibody

Cited by 11 publications

References 9 publications

Prevalence of African swine fever virus in apparently healthy domestic pigs in Uganda

Prevalence of African swine fever virus in apparently healthy domestic pigs in Uganda

The role of nuclear technologies in the diagnosis and control of livestock diseases—a review

Identification of the principal serological immunodeterminants of African swine fever virus by screening a virus cDNA library with antibody

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