African swine fever virus serodiagnosis: A general review with a focus on the analyses of African serum samples

Cubillos, Carolina; Gómez-Sebastián, Silvia; Moreno, Noelia; Núñez, María; Mulumba‐Mfumu, Léopold K.; Quembo, C; Heath, Livio; Etter, Éric; Jori, Ferrán; Escribano, José M.; Blanco, Esther

doi:10.1016/j.virusres.2012.10.021

Cited by 99 publications

(86 citation statements)

References 43 publications

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“…Therefore, the use of the most appropriate diagnostic tools that are updated to be applicable to all scenarios is critical for the implementation of effective control programs. Previous studies have examined the performance of serological (40,42) and virological (34) ASF diagnostic tests with a relatively large number of samples, which were mainly obtained from the African continent. The recent emergence of ASF in 2014 in some eastern EU countries has highlighted the need to assess ASF diagnostic tests to check their ability to detect the disease via viral DNA, antigens, and antibodies in clinical specimens in infected areas.…”

Section: Discussionmentioning

confidence: 99%

“…For the detection of ASF antibodies, the OIE-prescribed assays involve the use of an ELISA for antibody screening, backed up by immunoblotting (IB) or indirect immunofluorescence (IIF) as confirmatory tests (33,40). The indirect immunoperoxidase test (IPT), validated by the European Union Reference Laboratory (EURL) for ASF, has effective analytical and diagnostic sensitivity and can be used as an alternative confirmatory test for the diagnosis of ASF using porcine sera.…”

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confidence: 99%

“…The techniques currently in use for ASF diagnosis give a confident diagnosis of the disease in any epidemiological situation (34,35,40,42). However, ASF diagnosis is complex due to the wide range of clinical forms and the similarity of its symptoms to those of other viral infections, such as classical swine fever (CSF) (1).…”

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confidence: 99%

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Assessment of African Swine Fever Diagnostic Techniques as a Response to the Epidemic Outbreaks in Eastern European Union Countries: How To Improve Surveillance and Control Programs

et al. 2015

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A frican swine fever (ASF) is a complex and lethal viral disease affecting swine and has a significant socioeconomic impact on both the developed and developing world. It has a major negative effect on national, regional, and international trade and constrains pig production in affected areas. The devastating acute form of the disease is characterized, among other features, by functional and congestive-hemorrhagic disorders of the digestive and respiratory systems and causes around 100% mortality in infected pigs (1). Both European wild boars (Sus scrofa) and feral pigs are susceptible and exhibit clinical signs and mortality rates similar to those of domestic pigs. In contrast, African wild pigs (Phacochoerus and Potamochoerus spp.) are resistant to the disease (2-10).The causative agent of the disease, the ASF virus (ASFV), is a large double-stranded DNA virus and the only member of the Asfarviridae family, genus Asfivirus (11, 12). The virus genome is 170 to 192 kb long (13-17). ASF is endemic in sub-Saharan Africa, where it was first described in 1921 (18). Several outbreaks have occurred since then in Europe and South and Central America. In most non-African countries, the disease has been successfully eradicated, the only exception being Sardinia (Italy), where the disease is still endemic (19,20). In April 2007, the disease spread from East Africa to the Republic of Georgia (21), and outbreaks occurred in Armenia, Azerbaijan, and the Russian Federation (22). The ongoing spread of ASFV into adjacent eastern European countries, such as Ukraine (23,24) and Belarus (25), and the situation in Russia affecting both wild boars and domestic pigs placed neighboring areas in the European Union (EU) at risk for the spread of ASFV. The first cases of ASF in wild boars in Lithuania and Poland were reported in early 2014 in areas bordering . According to the World Organisation for Animal Health (OIE), during 2014, nearly 260 ASF cases or outbreaks in wild boars and domestic pigs were detected in EU countries (Latvia, Lithuania, Estonia, and Poland). This situation, combined with the uncertainty present in Belarus, has created a permanent risk of reintroducing ASF into the EU via wild boars or the illegal trade of contaminated pork products and waste (31).No vaccine is available to prevent ASF infection. The control and eradication measures applicable are based on classical disease control methods, including surveillance, epidemiological investigation, tracing of pigs, and stamping out in infected holdings. Citation Gallardo C, Nieto R, Soler A, Pelayo V, Fernández-Pinero J, MarkowskaDaniel I, Pridotkas G, Nurmoja I, Granta R, Simón A, Pérez C, Martín E, Fernández-Pacheco P, Arias M. 2015. Assessment of African swine fever diagnostic techniques as a response to the epidemic outbreaks in eastern European Union countries: how to improve surveillance and control programs.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Assessment of African Swine Fever Diagnostic Techniques as a Response to the Epidemic Outbreaks in Eastern European Union Countries: How To Improve Surveillance and Control Programs

et al. 2015

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“…So far, the biological function of ASFV pK205R is not completely clear, but immunofluorescence techniques showed that pK205R is increasingly expressed in Vero cells (Gutierrez-Castaneda et al 2008, Cubillos et al 2013, and pK205R can be detected dispersing in the cytoplasm through fluorescent microscopy 4 h after artificial infection of Vero cells (Cubillos et al 2013). Then pK205R is gathered near the cell nucleus, which suggested that pK205R participates in the assembly of virions (Gutierrez-Castaneda et al 2008).…”

Section: Introductionmentioning

confidence: 99%

Prokaryotic expression, purification and antigenicity analysis of African swine fever virus pK205R protein

Wu¹,

Lu²,

Peng³

et al. 2016

Polish Journal of Veterinary Sciences

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African swine fever is an acute, febrile and highly virulent porcine disease causing serious economic losses worldwide. The pK205R protein of the African swine fever virus (ASFV) is largely expressed in the early stages of infection, which has given the K205R gene extensive attention. In this study, the ASFV K205R was cloned and expressed in Escherichia coli BL21 (DE3). Expression of histidine-tagged pK205R with a molecular mass of 44 kDa was determined by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. Optimisation of culture conditions allowed induction of the recombinant protein with 0.4 mM Isopropyl β-D-thiogalactoside (IPTG) at 37 o C for 2 h. The protein existed in cellular supernatant and was purified using a Ni-NTA resin column. The purified protein was used to immunize rabbits four times to enable the production of polyclonal antibodies, and the antiserum titre was detected by ELISA. The results showed that the purified pK205R can react with ASFV positive serum specifically by Western blotting. The pK205R had high antigenicity, which indicated that pK205R could be used as an antigen for detection of ASFV-specific antibodies in ELISA testing, and the recombinant protein could contribute to further research of the action and structure of pK205R.

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“…In subacute and chronic disease, virus-specific antibodies are found in blood on days 7-10 [13][14][15][16], so earlier a detection of the viral antigen or genome in blood is advisable [17], since viremia is typical of this ASF period [17][18][19].…”

Section: Introductionmentioning

confidence: 99%

Validation of an Elisa Kit for Detection of Antibodies Against Asf Virus in Blood or Spleen of Domestic Pigs and Wild Boars

Strizhakova¹,

Lyska²,

Malogolovkin³

et al. 2016

S-h. biol.

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A b s t r a c tThe causal agent of African swine fever (ASF) is a DNA virus belonging to Asfaviridae family which affects both wild boar Sus scrofa and domestic pig Sus scrofa domestica. Special features of the course of (ASF and its forms should be considered for the effective use of various ASF diagnostic methods aimed at the pathogen or specific antibody identification. ASF diagnosis in the wild boar is of special importance. The wild boar susceptibility to ASF virus is well known, and the disease has been repeatedly reproduced in experiments and detected in European wild boars in natural conditions. It is not unfrequently that when shooting wild boars, only organ samples are delivered to laboratories, so we decided to estimate the diagnostic value of the antibodies detected in tissues and evaluate an earlier developed test system for its efficacy when used for the assay. This report represents the results of validation of an indirect ELISA (a commercial kit «VNIIVViM ASF-ELISA Ab/Ag») ASF virus specific antibody detection in blood serum and spleen tissue extracts. For comparison, an indirect immunofluorescence assay (indirect IFA) was used. To estimate the obtained results, ROC analysis was applied. Examination of positive (n = 66) and negative (n = 410) porcine blood serum samples using indirect ELISA showed high sensitivity and specificity of the method with reference to IFA. Among the 476 serum samples examined in indirect ELISA, only 8 sera (1.6 %) were within the positive/negative cutoff area. The highest sensitivity (100 %) and specificity (99.27 %) for indirect ELISA when examining both domestic pig and wild boar blood sera were determined at a cutoff value of 0.264. The samples of spleen extracts to be used for the study were collected from clinically healthy wild boars in ASF-affected Smolensk region in 2013 to 2014. In view of an acute form of the disease, we can suspect that the animals exhibiting positive reaction were in a latency period of the pathogeny. When examining the positive (n = 59) and negative (n = 678) spleen extracts, we also determined high sensitivity and specificity levels of indirect ELISA with reference to indirect IFA. Among the samples examined (n = 737), 10 (1.3 %) samples of spleen extracts were within the positive/negative cutoff area. The highest sensitivity (100 %) and specificity (98.82 %) were observed at a cutoff value of 0.284. Thus, we confirmed the assay to be effective for porcine blood sera with 100 % sensitivity (94.6 to 100 %) and 99.27 % specificity (97.90 to 99.80 %) and for spleen extracts with 100 % sensitivity (93.90 to 100 %) and 98.82 % specificity (97.70 to 99.50 %).

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African swine fever virus serodiagnosis: A general review with a focus on the analyses of African serum samples

Cited by 99 publications

References 43 publications

Assessment of African Swine Fever Diagnostic Techniques as a Response to the Epidemic Outbreaks in Eastern European Union Countries: How To Improve Surveillance and Control Programs

Assessment of African Swine Fever Diagnostic Techniques as a Response to the Epidemic Outbreaks in Eastern European Union Countries: How To Improve Surveillance and Control Programs

Prokaryotic expression, purification and antigenicity analysis of African swine fever virus pK205R protein

Validation of an Elisa Kit for Detection of Antibodies Against Asf Virus in Blood or Spleen of Domestic Pigs and Wild Boars

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