Serotype 4 pneumococcus
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ABSTRACT:
PROVIDER: PRJEB42817 | ENA |
REPOSITORIES: ENA
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Project description:We were interested in determining if the LuxS signaling system contributed to the regulation of pneumococcal genes. To accomplish this, we compared the in vitro transcriptional profiles over time of S. pneumoniae D39, a serotype 2 strain of pneumococcus, with that of an isogenic deletion mutant, (delta)luxS, using a spotted S. pneumoniae- specific DNA microarray. For in vitro time course experiments, broth cultures of D39 and (delta)luxS were grown to low density (optical densities between ~0.01- 0.02), the cells were collected by centrifugation, washed 1x with sterile PBS, back diluted to ~3x105CFU/ ml into fresh broth, and immediately incubated at 37oC in a 5% CO2 chamber for 45 minutes. At this point, the first sample was collected. Subsequently, growth was monitored for approximately 8 generations by measuring both the optical density (OD600) and determining viable counts every 30 to 45 minutes. At these points samples were also harvested by centrifugation for RNA isolation and stored at -80oC until processing. For microarray probe synthesis, experimental samples were generated using 0.5ug total bacterial RNA isolated from either D39 or (delta)luxS strains at the various time points throughout the growth curve. These RNAs were used as templates for reverse transcription, and the resulting cDNAs were labelled with Cy5. The reference sample for each time course was generated from the D39 time zero RNA. Subsequent to reverse transcription, the resulting cDNAs were labelled with Cy3. The probes were mixed in hybridization buffer, heated to 99oC for at least 2 minutes, centrifuged briefly, and applied to the microarray. Hybridizations were carried out at 60oC for at least 24 hours.
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Project description:Non-vaccine type penicillin nonsusceptible pneumococcus
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Project description:Streptococcus pneumoniae is a frequent coloniser of the human nasopharynx and a major cause of life-threating invasive infections such as pneumonia, meningitis and sepsis. Over 1 million people die every year due to invasive pneumococcal disease (IPD), mainly in developing countries. Serotype 1 is a common cause of IPD; however, unlike other serotypes, it is rarely found in the carrier state in the nasopharynx, which is often considered a prerequisite for disease. The aim of this study was to understand this dichotomy. We used murine models of carriage and IPD to characterise the pathogenesis of African serotype 1 (Sequence Type 217) pneumococcal strains obtained from the Queen Elizabeth Central Hospital in Blantyre, Malawi. We found that ST217 pneumococcal strains were highly virulent in a mouse model of invasive pneumonia, but in contrast to the generally accepted assumption, can also successfully establish nasopharyngeal carriage. Interestingly, we found that co-colonising serotypes may proliferate in the presence of serotype 1, suggesting that acquisition of serotype 1 carriage could increase the risk of developing IPD by other serotypes. RNAseq analysis confirmed that key virulence genes associated with inflammation and tissue invasiveness were upregulated in serotype 1. These data reveal important new insights into serotype 1 pathogenesis, with implications for carriage potential and risk of invasive disease through interactions with other co-colonising serotypes; an often overlooked factor in transmission and disease progression.
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Project description:We used RNA-Seq to compare gene expression between representative serotype M1 and M3 GAS isolates
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Project description:Streptococcus pneumoniae (the pneumococcus) account for significant morbidity and mortality worldwide, causing life-threatening diseases such as pneumonia, bacteremia and meningitis. In this study, we used microarray analysis to compare gene expression patterns of either serotype 4 or serotype 6A pneumococci in the nasopharynx and blood of mice, as a model to identify genes involved in invasion of blood in the context of occult bacteremia in humans.
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Project description:The capsular serotype has long been associated with the virulence of Streptococcus pneumoniae. Here we present an in-depth study of phenotypic and genetic differences between serotype 3 and serogroup 11 S. pneumoniae clinical isolates from both the general and indigenous populations of Australia. Both serotypes/groups included clonally unrelated strains with differences in well-known polymorphic virulence genes, such as nanA and pspA, as demonstrated by multilocus sequence typing and Western blot analysis. Nonetheless, the serotype 3 strains were consistently and significantly more virulent in mice than the serogroup 11 strains. Despite extensive genomic analysis, noncapsular genes common to one serotype/group but not the other were not identified. Nevertheless, following the conversion of a serotype 11A isolate to serotype 3 and subsequent analysis in an intranasal infection model, it was evident that both capsular and noncapsular factors determine the virulence phenotype in mice. However, it appears that these noncapsular factors vary from strain to strain. Data is also available from http://bugs.sgul.ac.uk/E-BUGS-126
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