Nonlinear Finite Element Analysis of Nanoindentation of Viral Capsids

Recent Atomic Force Microscope (AFM) nanoindentation experiments measuring mechanical response of the protein shells of viruses have provided a quantitative description of their strength and elasticity. To better understand and interpret these measurements, and to elucidate the underlying mechanisms, this paper adopts a course-grained modeling approach within the framework of three-dimensional nonlinear continuum elasticity. Homogeneous, isotropic, elastic, thick shell models are proposed for two capsids: the spherical Cowpea Chlorotic Mottle Virus (CCMV), and the ellipsocylindrical bacteriophage $\phi 29$. As analyzed by the finite element method, these models enable parametric characterization of the effects of AFM tip geometry, capsid dimensions, and capsid constitutive descriptions. The generally nonlinear force response of capsids to indentation is shown to be insensitive to constitutive details, and greatly influenced by geometry. Nonlinear stiffening and softening of the force response is dependent on the AFM tip dimensions and shell thickness. Fits of the models capture the roughly linear behavior observed in experimental measurements and result in estimates of Young's moduli of $\approx$280--360 MPa for CCMV and $\approx$4.5 GPa for $\phi 29$.Comment: 24 pages, 10 figures, submitted to Biophysical Journa

Teste imunoenzimático com base em anticorpo monoclonal para a detecção de anticorpos contra os herpesvírus bovino tipos 1 e 5

Os herpesvírus bovino tipos 1 (BoHV-1) e 5 (BoHV-5) são agentes virais genética e antigenicamente relacionados, associados com diversas manifestações clínicas em bovinos, incluindo doença respiratória, genital, neurológica e abortos. Estudos epidemiológicos indicam que esses vírus estão amplamente disseminados no rebanho bovino brasileiro. O diagnóstico sorológico, que permite identificar animais portadores da infecção latente, se constitui em importante ferramenta para monitoramento individual e de rebanho. O presente artigo relata a padronização de um teste imunoenzimático do tipo ELISA, com base em anticorpo monoclonal (AcM), para a detecção de anticorpos séricos que reagem contra BoHV-1 e/ou BoHV-5. Inicialmente, determinou-se o AcM mais adequado para a sensibilização das placas, as diluições apropriadas do antígeno e dos soros-teste e o ponto de corte do ensaio. Após a padronização, o ensaio foi validado testando-se 506 amostras de soro bovino, previamente testadas para anticorpos neutralizantes contra BoHV-1 e/ou BoHV-5 pela técnica de soroneutralização (SN). Comparando-se com os resultados da SN frente a BoHV-1, o teste de ELISA apresentou sensibilidade e especificidade de 96,6% e 98,3%, respectivamente. Os valores preditivos positivo e negativo foram de 97,6%, a concordância foi de 97,6% e o índice de correlação kappa entre os testes foi de 0,95, o que indica uma excelente concordância. Comparando-se com os resultados da SN frente o BoHV-5, o ELISA apresentou 94,3% de sensibilidade; 97,9% de especificidade; 97,1% de valor preditivo positivo e 95,9% de valor preditivo negativo. Para BoHV-5, a concordância entre os testes foi de 96,4% e o índice de correlação foi de 0,92, também excelente. Esses resultados demonstram que o teste padronizado apresenta sensibilidade e especificidade adequados para o diagnóstico sorológico das infecções por BoHV-1 e BoHV-5 em nível individual e de rebanho. Dessa forma, o ensaio pode se constituir em alternativa para o teste de SN e para os kits de ELISA importados.Bovine herpesviruses 1 (BoHV-1) and 5 (BoHV-5) are antigenic and genetically related viruses associated with different clinical syndromes in cattle, including respiratory, reproductive, neurological disease and abortion. Epidemiological studies indicate the widespread distribution of both viruses among Brazilian cattle. Serological diagnosis, that allows the identification of latently infected animals, represents an important tool for individual and herd monitoring. The present article describes the standardization of a monoclonal antibody (MAb)-based immunoenzymatic test (ELISA) for detection of antibodies to BoHV-1 and/or BoHV-5. The initial steps involved the determination of the most suitable MAb, the appropriate dilutions of viral antigen and serum samples, and the cut-off value of the assay. After standardization, the ELISA was validated by testing 506 cattle serum samples previously tested for neutralizing antibodies to BoHV-1 and BoHV-5 by virus neutralizing assay (VN). Comparing to the VN for BoHV-1 antibodies, the ELISA presented sensitivity and specificity of 96.6% and 98.3%, respectively. Positive and negative predictive values were 97.6%, the concordance between the tests was 97.6% and the coefficient of correlation k (kappa) was 0.95, demonstrating an excellent correlation. Comparing to the VN for BoHV-5 antibodies, the ELISA presented 94.3% of sensitivity, 97.9% of specificity, 97.1% of positive predictive value, 95.9% negative predictive value, concordance of 96.4% and kappa coefficient of 0.92. These results demonstrate that the ELISA presents suitable specificity and sensitivity to be used for individual and herd serological diagnosis of BoHV-1 and BoHV-5, thus, representing an alternative for VN assays and imported ELISA kits

De structurele menselijke ooglenseiwitten en seniele nucleaire cataract

Contains fulltext : mmubn000001_207009848.pdf (publisher's version ) (Open Access)Promotores : H. Hoenders en H. Bloemendal117 p

Critical factors affecting the diagnostic reliability of enzyme-linked immunosorbent assay formats

This paper aims to evaluate different formats of the enzyme-linked immunosorbent assays (ELISAs) for detection of virus-specific antibodies and focuses on factors that may influence the diagnostic reliability of such tests. Newly developed and well-established ELISAs for detection of infections of bovine herpesvirus 1 (BHV1), bovine respiratory syncytial virus (BRSV), classical swine fever virus (CSFV), pseudorabies virus (PRV) and bovine viral diarrhoea virus (BVDV) are used as examples. Differences between competitive and non-competitive ELISAs are described, with special reference to the influence of the antigen, the conjugated antibody and the test sample on the test results. Attention is drawn to interference, which may result in false positive or false negative test results, with special emphasis on the 'bridging' phenomenon. The use of monoclonal antibodies and discriminatory tests are briefly discussed. Diagnostic reliability is described for tests that are used in monitoring or eradication programmes, emphasising the consequences of false negative and false positive test results. Finally, reducing assay-time and functional quality control for such tests are discussed

Critical factors affecting the diagnostic reliability of enzyme-linked immunosorbent assay formats

This paper aims to evaluate different formats of the enzyme-linked immunosorbent assays (ELISAs) for detection of virus-specific antibodies and focuses on factors that may influence the diagnostic reliability of such tests. Newly developed and well-established ELISAs for detection of infections of bovine herpesvirus 1 (BHV1), bovine respiratory syncytial virus (BRSV), classical swine fever virus (CSFV), pseudorabies virus (PRV) and bovine viral diarrhoea virus (BVDV) are used as examples. Differences between competitive and non-competitive ELISAs are described, with special reference to the influence of the antigen, the conjugated antibody and the test sample on the test results. Attention is drawn to interference, which may result in false positive or false negative test results, with special emphasis on the 'bridging' phenomenon. The use of monoclonal antibodies and discriminatory tests are briefly discussed. Diagnostic reliability is described for tests that are used in monitoring or eradication programmes, emphasising the consequences of false negative and false positive test results. Finally, reducing assay-time and functional quality control for such tests are discussed

Public health awareness of emerging zoonotic viruses of bats: A European perspective

Bats classified in the order Chiroptera are the most abundant and widely distributed non-human mammalian species in the world. Several bat species are reservoir hosts of zoonotic viruses and therefore can be a public health hazard. Lyssaviruses of different genotypes have emerged from bats in America (Genotype 1 rabies virus; RABV), Europe (European bat lyssavirus; EBLV), and Australia (Australian bat lyssavirus; ABLV), whereas Nipah virus is the most important recent zoonosis of bat origin in Asia. Furthermore, some insectivorous bat species may be important reservoirs of SARS coronavirus, whereas Ebola virus has been detected in some megachiropteran fruit bats. Thus far, European bat lyssavirus (EBLV) is the only zoonotic virus that has been detected in bats in Europe. New zoonotic viruses may emerge from bat reservoirs and known ones may spread to a wider geographical range. To assess future threats posed by zoonotic viruses of bats, there is a need for accurate knowledge of the factors underlying disease emergence, for an effective surveillance programme, and for a rapid response system. In Europe, primary efforts should be focussed on the implementation of effective passive and active surveillance systems for EBLVs in the Serotine bat, Eptesicus serotinus, and Myotis species (i.e., M. daubentonii and M. dasycneme) Apart from that, detection methods for zoonotic viruses that may emerge from bats should be implemented. Analyses of data from surveillance studies can shed more light on the dynamics of bat viruses, (i.e., population persistence of viruses in bats). Subsequently, studies will have to be performed to assess the public health hazards of such viruses (i.e., infectivity and risk of infection to people). With the knowledge generated from this kind of research, a rapid response system can be set up to enhance public health awareness of emerging zoonotic viruses of bats