Padronização do teste ELISA baseado em antígeno capsular purificado dos sorotipos 3, 5 e 7 de Actinobacillus pleuropneumoniae Standarization of ELISA test based on purified capsular antigen from serotypes 3, 5 and 7 of Actinobacillus pleuropneumoniae

Foram padronizados testes de ELISA (Enzyme-linked immunosorbent assay) baseados em antígeno capsular purificado de Actinobacillus pleuropneumoniae sorotipos 3, 5 e 7, prevalentes no Brasil. Para a padronização foram utilizadas amostras de soro provenientes de leitões inoculados com os três sorotipos do agente em estudo, dos quais se colheram amostras de sangue semanais, durante 15 semanas para estudo da dinâmica da síntese de anticorpos. O controle negativo dos testes constituiu-se de um mistura de 130 soros de animais livres de Actinobacillus pleuropneumoniae (App). Os antígenos também foram testados com amostras de soro de animais infectados com outros agentes causadores de doenças respiratórias e vacinados contra rinite atrófica. Os antígenos produzidos foram eficientes na detecção de animais infectados com App, permitindo determinar densidades óticas superiores à média dos soros controles negativos acrescida de quatro desvios-padrões. Os testes de ELISA para os sorotipos 3, 5 e 7 apresentaram especificidade de 100% e sensibilidade de 92, 88 e 90%, respectivamente. Não ocorreram reações cruzadas com outros sorotipos, assim como com soros de animais inoculados com outros agentes causadores de problemas respiratórios. Os resultados foram analisados através da análise discriminante de ANDERSON (1958), utilizando-se o programa Statistical Analysis System. Concluiu-se que os antígenos testados são adequados para sorotipar animais que tenham sido submetidos ao screening através de um teste de ELISA polivalente baseado em LPS-LC.<br>Three ELISA (Enzime-linked immunosorbent assay) tests based on purified capsular antigen from serotypes 3, 5 and 7 of Actinobacillus pleuropneumoniae, prevalent in Brazil, were standardized. Serum samples, collected from piglets inoculated with these three serotypes, were used to standardize the test. In order to study the dynamic of antibody synthesis, weekly blood samples were collected from these piglets. A pool of 130 sera obtained from Actinobacillus pleuropneumoniae -free pigs was used as negative control for the tests. The antigens were also tested with serum samples from animals infected with other respiratory infectious agents and vaccinated against athrofic rinitis. The antigens were efficient in detecting animals infected with App. Optical densities above the average of the negative control sera plus four standard deviation were detected. ELISA tests to serotypes 3, 5 and 7 showed specificity of 100% and sensibility of 92, 88 and 90%, respectively. No cross reaction with other serotypes or with sera of animals inoculated with other respiratory pathogens was observed. The results were analyzed using the Statistical Analysis System program. Antigens tested were adequate for serotyping animals previously screened through a polyvalent LPS-LC ELISA test

Pan-cancer analysis of whole genomes

Cancer is driven by genetic change, and the advent of massively parallel sequencing has enabled systematic documentation of this variation at the whole-genome scale. Here we report the integrative analysis of 2,658 whole-cancer genomes and their matching normal tissues across 38 tumour types from the Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium of the International Cancer Genome Consortium (ICGC) and The Cancer Genome Atlas (TCGA). We describe the generation of the PCAWG resource, facilitated by international data sharing using compute clouds. On average, cancer genomes contained 4-5 driver mutations when combining coding and non-coding genomic elements; however, in around 5% of cases no drivers were identified, suggesting that cancer driver discovery is not yet complete. Chromothripsis, in which many clustered structural variants arise in a single catastrophic event, is frequently an early event in tumour evolution; in acral melanoma, for example, these events precede most somatic point mutations and affect several cancer-associated genes simultaneously. Cancers with abnormal telomere maintenance often originate from tissues with low replicative activity and show several mechanisms of preventing telomere attrition to critical levels. Common and rare germline variants affect patterns of somatic mutation, including point mutations, structural variants and somatic retrotransposition. A collection of papers from the PCAWG Consortium describes non-coding mutations that drive cancer beyond those in the TERT promoter; identifies new signatures of mutational processes that cause base substitutions, small insertions and deletions and structural variation; analyses timings and patterns of tumour evolution; describes the diverse transcriptional consequences of somatic mutation on splicing, expression levels, fusion genes and promoter activity; and evaluates a range of more-specialized features of cancer genomes