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ABSTRACT: Foot and mouth disease (FMD) is a highly infectious viral disease that affects cattle, sheep, goats and swine causing severe economic losses worldwide. The efficacy of inactivated vaccines is critically dependent on the integrity of foot and mouth disease virus (FMDV) particles. The recommended method to quantify the active ingredient of vaccines is the 140S quantitative sucrose density gradient analysis. This method has been an immensely valuable tool over the past three decades but it is highly operator dependent and difficult to automate. We developed a method to quantify FMDV particles during the vaccine manufacturing process that is based on separation of components by size-exclusion chromatography and measurement of virus by absorption at 254nm. The method is linear in the 5-70μg/mL range, it is applicable to different FMDV strains, and has a good correlation with the 140S test. The proposed method uses standard chromatographic media and it is amenable to automation. The method has potential as a process analytical technology and for control of final product by manufacturers, international vaccine banks and regulatory agencies.
Vaccine 06/2011; 29(41):7182-7. · 3.77 Impact Factor
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ABSTRACT: The World Organisation for Animal Health (OIE) Terrestrial Manual and the European Pharmacopoeia (EP) still prescribe live challenge experiments for foot-and-mouth disease virus (FMDV) immunogenicity and vaccine potency tests. However, the EP allows for other validated tests for the latter, and specifically in vitro tests if a "satisfactory pass level" has been determined; serological replacements are also currently in use in South America. Much research has therefore focused on validating both ex vivo and in vitro tests to replace live challenge. However, insufficient attention has been given to the sensitivity and specificity of the "gold standard"in vivo test being replaced, despite this information being critical to determining what should be required of its replacement. This paper aims to redress this imbalance by examining the current live challenge tests and their associated statistics and determining the confidence that we can have in them, thereby setting a standard for candidate replacements. It determines that the statistics associated with the current EP PD(50) test are inappropriate given our domain knowledge, but that the OIE test statistics are satisfactory. However, it has also identified a new set of live animal challenge test regimes that provide similar sensitivity and specificity to all of the currently used OIE tests using fewer animals (16 including controls), and can also provide further savings in live animal experiments in exchange for small reductions in sensitivity and specificity.
Vaccine 06/2011; 29(33):5467-73. · 3.77 Impact Factor
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Blanca Robiolo,
José La Torre,
Eduardo Maradei,
Claudia Perez Beascoechea,
Alejandro Perez,
Cristina Seki, Eliana Smitsaart,
Norberto Fondevila,
Eduardo Palma,
Nesya Goris,
Kris De Clercq,
Nora Mattion
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ABSTRACT: The necessity of avoiding the use of animals in vaccine potency testing has been widely recognized. The repeatability and reproducibility of the Expected Percentage of Protection (EPP) as a serological potency surrogate for A24 Cruzeiro foot-and-mouth disease virus (FMDV) strain was assessed, and compared with the results obtained with challenge in the Protection against Podal Generalization (PPG) test. To determine the EPPs, the serum titers obtained by liquid phase blocking competitive ELISA (lpELISA) and virus neutralization (VNT) in 10 potency trials using the same A24 Cruzeiro vaccine, were interpolated into previously validated logit transformation curves that correlate PPG with serology. Indirect serological assessment of vaccine matching between the serotype A FMDV strains A24 Cruzeiro and A/Argentina/01 was also carried out by lpELISA and VNT. The results obtained in this study strongly support the replacement of challenge tests for vaccine potency by indirect serological assays, at least for A24 Cruzeiro FMDV strain. While determination of EPPs by lpELISA titers showed an excellent repeatability, reproducibility and concordance with PPG for vaccine potency, assessments of cross-protection by VNT titers were more consistent with the PPG outcome.
Vaccine 08/2010; 28(38):6235-41. · 3.77 Impact Factor
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Nora Mattion,
Nesya Goris,
Tom Willems,
Blanca Robiolo,
Eduardo Maradei,
Claudia Perez Beascoechea,
Alejandro Perez, Eliana Smitsaart,
Norberto Fondevila,
Eduardo Palma,
Kris De Clercq,
José La Torre
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ABSTRACT: The selection of matching strains for use in outbreaks of foot-and-mouth disease (FMD) virus can be assessed in vivo or by serological r-value determination. Sera from animals involved in vaccine potency and cross-protection trials performed using the "Protection against Podal Generalization" (PPG) test for two serotype A strains were collected and analyzed by the virus neutralization test (VNT) and liquid-phase ELISA (lpELISA) in three laboratories. The average VNT r-values for medium and high serum titer classes from the A(24) Cruzeiro vaccinated animals were in line with the A/Arg/01 heterologous PPG outcome for all testing laboratories, suggesting that the vaccine strain A(24) Cruzeiro is unlikely to protect against the field isolate A/Arg/01. The corresponding lpELISA r-values were slightly higher and indicate a closer relationship between both strains. Pooling of serum samples significantly reduced the inter-animal and inter-trial variation. The results suggest that a suitable reference serum for vaccine matching r-value experiments might be a pool or a medium to high VNT or lpELISA titer serum. Furthermore, the VNT seems to produce the most reproducible inter-laboratory results. More work is, however, needed in order to substantiate these claims.
Vaccine 12/2008; 27(5):741-7. · 3.77 Impact Factor
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ABSTRACT: The use of foot-and-mouth disease (FMD) vaccines that do not induce antibodies against non-structural proteins (NSP) is extremely relevant for the demonstration of regions "free of FMDV infection" and control strategies. In this study cattle were primed and boosted with five doses of oil vaccines containing high antigenic payloads on days 0, 90, 130, 160 and 200. The serological response against NSP was measured using four commercially available assays: two 3ABC-ELISAs; one 3B-ELISA (and complementary 3A-ELISA) and an enzyme-immunotransfer blot assay (EITB). Additionally, locally produced NSP antibodies detection reagents and VIAA antibodies were evaluated. A high level of specific immune response against vaccine strains was shown. After four doses of vaccine, non-reactive animals were detected by any of the NSP assays. After the fifth immunization, 2 of 17 animals were reactive in one ELISA kit, but these samples proved negative by confirmatory tests. Antibodies against NSP were not detected in single dose immunized cattle. The principle of the NSP-ELISA used as a screening test for large sero-surveys in South America is established and this paper emphasizes the importance of using vaccines that have demonstrated no interference with NSP antibodies detection assays.
Vaccine 12/2004; 23(1):69-77. · 3.77 Impact Factor
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Nora Mattion,
Guido König,
Cristina Seki, Eliana Smitsaart,
Eduardo Maradei,
Blanca Robiolo,
Sergio Duffy,
Emilio León,
María Piccone,
Ana Sadir,
Rodolfo Bottini,
Bernardo Cosentino,
Abraham Falczuk,
Ricardo Maresca,
Osvaldo Periolo,
Rodolfo Bellinzoni,
Ana Espinoza,
José La Torre,
Eduardo L Palma
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ABSTRACT: This paper describes the antigenic and molecular characterisation of foot-and-mouth disease virus (FMDV) strains isolated during the 2000-2002 epidemic in Argentina, and the strategy implemented for disease control. Two different FMDV serotypes, O and A, were involved. Of the various field isolates studied, two distinct O1 lineages (strains Corrientes/00 and Misiones/00) and two serotype A lineages (A/Argentina/00 and A/Argentina/01 prototypes) were identified. The genome sequences of these strains were compared with sequences of previous regional isolates and sequences of vaccine strains. O1 strains were found to be related to regional strains while serotype A strains were found to be more distanced from them. The updating of the antigenic composition of the vaccines used in the emergency was a key issue, since the outbreaks stopped shortly after the implementation of the vaccination programs. The O1 strains quickly disappeared from the field following strict control measures and the use of vaccines containing O1/Campos strain. However, in the case of the A serotype strains, the situation was different, since the use of a vaccine containing strain A24/Cruzeiro yielded acceptable levels of protection only after re-vaccination. Therefore, the new field strains A/Argentina/00 and A/Argentina/01 were incorporated into the vaccine, leading to an effective control of the disease. Viral circulation greatly diminished, as indicated by the significant reduction in the number of outbreaks and in the number of animals with antibodies against non-structural proteins. Satisfactory levels of protective antibodies were subsequently detected in the cattle population (above 75% protection). The absence of outbreaks after January 2002 indicated that the epidemic was controlled.
Vaccine 11/2004; 22(31-32):4149-62. · 3.77 Impact Factor
