Prospective, population-based surveillance of the burden of Streptococcus pneumoniae in community-acquired pneumonia in older adults, Chrzanów County, Poland, 2010 to 2012

INTRODUCTION: Community-acquired pneumonia (CAP) caused by Streptococcus pneumoniae is a substantial cause of morbidity and mortality among older adults. This study estimated incidences of CAP, chest x-ray−confirmed CAP (CXR+CAP), S pneumonia- positive CAP, S pneumonia-positive CXR+CAP, and S. pneumoniae serotype distribution among 46,000 at-risk adults aged ≥ 50 years residing in Chrzanów County, Poland. MATERIAL AND METHODS: From January 2010 to January 2012, all facilities providing ambulatory and inpatient care enrolled all consenting resident patients with suspicion of CAP. Chest x-rays, urine, blood, and sputum samples were analyzed. Annualized incidence rates were determined. Presence of S pneumonia-positive CAP and/or S. pneumoniae serotype distribution was determined using the urine antigen detection assay (capable of detecting the serotypes in the 13-valent pneumococcal conjugate vaccine [PCV13]), BinaxNOW®, and/or microbiology cultures. RESULTS: Among 5055 enrolled patients, 1195 (23.7%) were diagnosed with CAP and 1166 (23.4%) had CXR+CAP. S. pneumoniae was detected in 144 (12.1%) and 131 (11.2%) patients from the CAP and CXR+CAP cohorts, respectively. Annualized incidence rates of CAP, CXR+CAP, S pneumonia-positive CAP, and S. pneumonia-positive CXR+CAP were 12.8, 12.5, 1.6, and 1.4 per 1000 residents, respectively. Among CXR+CAP patients, 39.7% were aged 50 to 64 years and 60.3% were aged ≥ 65 years. Incidence rates generally increased with age. The most common serotypes in S. pneumoniae-positive CXR+CAP patients were 3 (n = 15), 23F (n = 10), 18C (n = 9), and 9V (n = 6). CONCLUSIONS: CAP due to PCV13 serotypes is a source of morbidity among adults >50 years and may be reduced by greater access to pneumococcal vaccines.INTRODUCTION: Community-acquired pneumonia (CAP) caused by Streptococcus pneumoniae is a substantial cause of morbidity and mortality among older adults. This study estimated incidences of CAP, chest x-ray−confirmed CAP (CXR+CAP), S pneumonia- positive CAP, S pneumonia-positive CXR+CAP, and S. pneumoniae serotype distribution among 46,000 at-risk adults aged ≥ 50 years residing in Chrzanów County, Poland. MATERIAL AND METHODS: From January 2010 to January 2012, all facilities providing ambulatory and inpatient care enrolled all consenting resident patients with suspicion of CAP. Chest x-rays, urine, blood, and sputum samples were analyzed. Annualized incidence rates were determined. Presence of S pneumonia-positive CAP and/or S. pneumoniae serotype distribution was determined using the urine antigen detection assay (capable of detecting the serotypes in the 13-valent pneumococcal conjugate vaccine [PCV13]), BinaxNOW®, and/or microbiology cultures. RESULTS: Among 5055 enrolled patients, 1195 (23.7%) were diagnosed with CAP and 1166 (23.4%) had CXR+CAP. S. pneumoniae was detected in 144 (12.1%) and 131 (11.2%) patients from the CAP and CXR+CAP cohorts, respectively. Annualized incidence rates of CAP, CXR+CAP, S pneumonia-positive CAP, and S. pneumonia-positive CXR+CAP were 12.8, 12.5, 1.6, and 1.4 per 1000 residents, respectively. Among CXR+CAP patients, 39.7% were aged 50 to 64 years and 60.3% were aged ≥ 65 years. Incidence rates generally increased with age. The most common serotypes in S. pneumoniae-positive CXR+CAP patients were 3 (n = 15), 23F (n = 10), 18C (n = 9), and 9V (n = 6). CONCLUSIONS: CAP due to PCV13 serotypes is a source of morbidity among adults >50 years and may be reduced by greater access to pneumococcal vaccines

Molecular epidemiology and expression of capsular polysaccharides in Staphylococcus aureus clinical isolates in the United States.

Staphylococcus aureus capsular polysaccharides (CP) are important virulence factors under evaluation as vaccine antigens. Clinical S. aureus isolates have the biosynthetic capability to express either CP5 or CP8 and an understanding of the relationship between CP genotype/phenotype and S. aureus epidemiology is valuable. Using whole genome sequencing, the clonal relatedness and CP genotype were evaluated for disease-associated S. aureus isolates selected from the Tigecycline Evaluation and Surveillance Trial (T.E.S.T) to represent different geographic regions in the United States (US) during 2004 and 2009-10. Thirteen prominent clonal complexes (CC) were identified, with CC5, 8, 30 and 45 representing >80% of disease isolates. CC5 and CC8 isolates were CP type 5 and, CC30 and CC45 isolates were CP type 8. Representative isolates from prevalent CC were susceptible to in vitro opsonophagocytic killing elicited by anti-CP antibodies, demonstrating that susceptibility to opsonic killing is not linked to the genetic lineage. However, as not all S. aureus isolates may express CP, isolates representing the diversity of disease isolates were assessed for CP production. While approximately 35% of isolates (primarily CC8) did not express CP in vitro, CP expression could be clearly demonstrated in vivo for 77% of a subset of these isolates (n = 20) despite the presence of mutations within the capsule operon. CP expression in vivo was also confirmed indirectly by measuring an increase in CP specific antibodies in mice infected with CP5 or CP8 isolates. Detection of antigen expression in vivo in relevant disease states is important to support the inclusion of these antigens in vaccines. Our findings confirm the validity of CP as vaccine targets and the potential of CP-based vaccines to contribute to S. aureus disease prevention

The Gonococcal Fur Regulon: Identification of Additional Genes Involved in Major Catabolic, Recombination, and Secretory Pathways

In this study, we have characterized the in vitro binding of Neisseria gonorrhoeae Fur to several well-defined iron transport genes, as well as to additional genes involved in major catabolic, secretory, and recombination pathways of gonococci. The gonococcal Fur protein was recombinantly expressed in Escherichia coli HBMV119. Fur was isolated from inclusion bodies and partially purified by ion-exchange chromatography. Gonococcal Fur was found to bind to the promoter/operator region of a gene encoding the previously identified Fur-regulated periplasmic binding protein (FbpA) in a metal ion-dependent fashion, demonstrating that purified Fur is functional. In silico analysis of the partially completed gonococcal genome (FA1090) identified Fur boxes in the promoters of several genes, including tonB, fur, recN, secY, sodB, hemO, hmbR, fumC, a hypothetical gene (Fe-S homolog), and the opa family of genes. By using purified gonococcal Fur, we demonstrate binding to the operator regions of tonB, fur, recN, secY, sodB, hemO, hmbR, fumC, the Fe-S homolog gene, and the opa gene family as determined by an electrophoretic mobility shift assay. While gonococcal Fur was demonstrated to bind to the promoter regions of all 11 opa genes (opaA through -K), we did not detect binding of purified E. coli Fur with 8 of the 11 opa members, indicating that target DNA sequence specificities between these two closely related proteins exist. Furthermore, we observed differences in the relative strengths of binding of gonococcal Fur for these different genes, which most likely reflect a difference in affinity between gonococcal Fur and its DNA targets. This is the first report that definitively demonstrates the binding of gonococcal Fur to its own promoter/operator region, as well as to the opa family of genes that encode surface proteins. Our results demonstrate that the gonococcal Fur protein binds to the regulatory regions of a broad array of genes and indicates that the gonococcal Fur regulon is larger than originally proposed

Expression of the Gonococcal Global Regulatory Protein Fur and Genes Encompassing the Fur and Iron Regulon during In Vitro and In Vivo Infection in Women▿ †

The ferric uptake regulatory protein, Fur, functions as a global regulatory protein of gene transcription in the mucosal pathogen Neisseria gonorrhoeae. We have shown previously that several N. gonorrhoeae Fur-repressed genes are expressed in vivo during mucosal gonococcal infection in men, which suggests that this organism infects in an iron-limited environment and that Fur is expressed under these conditions. In this study we have demonstrated expression of the gonococcal fur gene in vitro, in human cervical epithelial cells, and in specimens from female subjects with uncomplicated gonococcal infection. In vitro studies confirmed that the expression of the gonococcal fur gene was repressed during growth under iron-replete growth conditions but that a basal level of the protein was maintained. Using GFP transcriptional fusions constructed from specific Fur binding sequences within the fur promoter/operator region, we determined that this operator region was functional during N. gonorrhoeae infection of cervical epithelial cells. Furthermore, reverse transcription-PCR analysis, as well as microarray analysis, using a custom Neisseria Fur and iron regulon microarray revealed that several Fur- and iron-regulated genes were expressed during N. gonorrhoeae infection of cervical epithelial cells. Microarray analysis of specimens obtained from female subjects with uncomplicated gonococcal infection corroborated our in vitro findings and point toward a key role of gonococcal Fur- and iron-regulated genes in gonococcal disease

Prospektywna, populacyjna obserwacja występowania Streptococcus pneumoniae w przebiegu pozaszpitalnego zapalenia płuc wśród osób starszych, Chrzanów, Polska, 2010−2012

WSTĘP: Pozaszpitalne zapalenie płuc wywołane przez S. pneumoniae jest istotną przyczyną chorobowości i śmiertelności wśród osób starszych. Badanie to ocenia występowanie pozaszpitalnego zapalenia płuc (CAP, community acquired pneumonia), CAP potwierdzonego radiologicznie (CXR+CAP), S. pneumoniae dodatniego CAP, S. pneumoniae dodatniego CAP potwierdzonego radiologicznie oraz dystrybucję serotypów S. pneumoniae u ponad 46 000 dorosłych powyżej 50. roku życia mieszkających w powiecie chrzanowskim w Polsce. MATERIAŁ I METODY: Od stycznia 2010 do stycznia 2012 roku wszystkie placówki zapewniające amulatoryjną i szpitalną opiekę medyczną włączały wszystkich wyrażających zgodę pacjentów z podejrzeniem CAP. Wykonywano zdjęcie RTG klatki piersiowej, badanie moczu, krwi i plwociny. Uzyskano roczne wskażniki zapadalności. Potwierdzenie S. pneumoniae-dodatniego CAP i/lub rozkład serotypów S. pneumoniae uzyskiwano w analizie antygenowej moczu (BINAX NOW®) (wykrywający serotypy obecne w 13-walentnej szczepionce PCV 13) i posiewach mikrobiologicznych. WYNIKI: Wśród 5055 pacjentów włączonych do badania u 1195 (23,7%) zdiagnozowano pozaszpitalne zapalenie płuc, w tym 1166 (23,4%) miało zmiany radiologiczne (CAP+CXR). S. pneumoniae stwierdzono u 144 pacjentów z CAP (12,1%) i u 133 w grupie z CAP+CXR (11,2%). Roczne wskaźniki zapadalności na CAP, CAP+CXR, S. pneumonia-dodatniego CAP oraz S. pneumonia-dodatniego CAP+CXR wynosiły odpowiednio 12,8, 12,5, 1,6 i 1,4 na 1000 mieszkańców. Wśród pacjentów z CAP+CXR 39,7% było w grupie wiekowej 50−64, a 60,3% miało ≥ 65 lat. Częstość występowania rosła z wiekiem. Najczęstsze serotypy S. pneumoniae u pacjentów z CAP+CXR to 3 (n = 15), 23F (n = 10), 18C (n = 9), 9V (n = 6). WNIOSKI: CAP spowodowane serotypami PCV13 jest przyczyną chorobowości u dorosłych powyżej 50. roku życia, która może być zredukowana większym dostępem do szczepionki.WSTĘP: Pozaszpitalne zapalenie płuc wywołane przez S. pneumoniae jest istotną przyczyną chorobowości i śmiertelności wśród osób starszych. Badanie to ocenia występowanie pozaszpitalnego zapalenia płuc (CAP, community acquired pneumonia), CAP potwierdzonego radiologicznie (CXR+CAP), S. pneumoniae dodatniego CAP, S. pneumoniae dodatniego CAP potwierdzonego radiologicznie oraz dystrybucję serotypów S. pneumoniae u ponad 46 000 dorosłych powyżej 50. roku życia mieszkających w powiecie chrzanowskim w Polsce. MATERIAŁ I METODY: Od stycznia 2010 do stycznia 2012 roku wszystkie placówki zapewniające amulatoryjną i szpitalną opiekę medyczną włączały wszystkich wyrażających zgodę pacjentów z podejrzeniem CAP. Wykonywano zdjęcie RTG klatki piersiowej, badanie moczu, krwi i plwociny. Uzyskano roczne wskażniki zapadalności. Potwierdzenie S. pneumoniae-dodatniego CAP i/lub rozkład serotypów S. pneumoniae uzyskiwano w analizie antygenowej moczu (BINAX NOW®) (wykrywający serotypy obecne w 13-walentnej szczepionce PCV 13) i posiewach mikrobiologicznych. WYNIKI: Wśród 5055 pacjentów włączonych do badania u 1195 (23,7%) zdiagnozowano pozaszpitalne zapalenie płuc, w tym 1166 (23,4%) miało zmiany radiologiczne (CAP+CXR). S. pneumoniae stwierdzono u 144 pacjentów z CAP (12,1%) i u 133 w grupie z CAP+CXR (11,2%). Roczne wskaźniki zapadalności na CAP, CAP+CXR, S. pneumonia-dodatniego CAP oraz S. pneumonia-dodatniego CAP+CXR wynosiły odpowiednio 12,8, 12,5, 1,6 i 1,4 na 1000 mieszkańców. Wśród pacjentów z CAP+CXR 39,7% było w grupie wiekowej 50−64, a 60,3% miało ≥ 65 lat. Częstość występowania rosła z wiekiem. Najczęstsze serotypy S. pneumoniae u pacjentów z CAP+CXR to 3 (n = 15), 23F (n = 10), 18C (n = 9), 9V (n = 6). WNIOSKI: CAP spowodowane serotypami PCV13 jest przyczyną chorobowości u dorosłych powyżej 50. roku życia, która może być zredukowana większym dostępem do szczepionki

Prospective, Population-Based Surveillance of the Burden of Streptococcus pneumoniae in Community-Acquired Pneumonia in Older Adults, Chrzanów County, Poland, 2010 to 2012

Introduction: Community-acquired pneumonia (CAP) caused by Streptococcus pneumoniae is a substantial cause of morbidity and mortality among older adults. This study estimated incidences of CAP, chest X-ray−confirmed CAP (CXR+CAP), S pneumoniae-positive CAP, S pneumoniae-positive CXR+CAP, and S. pneumoniae serotype distribution among 46,000 at-risk adults aged ≥ 50 years residing in Chrzanów County, Poland. Material and methods: From January 2010 to January 2012, all facilities providing ambulatory and inpatient care enrolled all consenting resident patients with suspicion of CAP. Chest X-rays, urine, blood, and sputum samples were analyzed. Annualized incidence rates were determined. Presence of S pneumoniae-positive CAP and/or S. pneumoniae serotype distribution was determined using the urine antigen detection assay (capable of detecting the serotypes in the 13-valent pneumococcal conjugate vaccine [PCV13]), BinaxNOW®, and/or microbiology cultures. Results: Among 5055 enrolled patients, 1195 (23.7%) were diagnosed with CAP and 1166 (23.4%) had CXR+CAP. S. pneumoniae was detected in 144 (12.1%) and 131 (11.2%) patients from the CAP and CXR+CAP cohorts, respectively. Annualized incidence rates of CAP, CXR+CAP, S pneumoniae-positive CAP, and S. pneumoniae-positive CXR+CAP were 12.8, 12.5, 1.6, and 1.4 per 1000 residents, respectively. Among CXR+CAP patients, 39.7% were aged 50 to 64 years and 60.3% were aged ≥ 65 years. Incidence rates generally increased with age. The most common serotypes in S. pneumoniae-positive CXR+CAP patients were 3 (n = 15), 23F (n = 10), 18C (n = 9), and 9V (n = 6). Conclusions: CAP due to PCV13 serotypes is a source of morbidity among adults >50 years and may be reduced by greater access to pneumococcal vaccines

Development and Validation of 13-plex Luminex-Based Assay for Measuring Human Serum Antibodies to Streptococcus pneumoniae Capsular Polysaccharides

The pneumococcal enzyme-linked immunosorbent assay (ELISA) measures IgG antibodies in human serum, and it is an important assay that supports licensure of pneumococcal vaccines. The immune correlate of protection, 0.35 µg/ml of IgG antibodies, was determined by the ELISA method. Pfizer has developed a new Luminex-based assay platform to replace the ELISA. These papers describe the important work of (i) validating the Luminex-based assay and (ii) bridging the immune correlate of protection (0.35 µg/ml IgG) to equivalent values reported by the Luminex platform.A Luminex-based direct immunoassay (dLIA) platform has been developed to replace the standardized pneumococcal enzyme-linked immunosorbent assay platform. The multiplex dLIA simultaneously measures the concentration of serum immunoglobulin G (IgG) antibodies specific for pneumococcal capsular polysaccharide (PnPS) serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F. The assay uses poly-l-lysine (PLL)-conjugated PnPS, chemically coupled to spectrally distinct Luminex microspheres. Assay validation experiments were performed using residual human serum samples obtained from 13-valent pneumococcal conjugate vaccine (13vPnC) clinical studies. Assay results are expressed as IgG antibody concentrations in micrograms per milliliter using the international reference serum, 007sp. The lower limit of quantitation (LLOQ) for all serotypes covered in the 13-plex dLIA fell within the range of 0.002 to 0.038 µg/ml serum IgG. The difference between the lower limit and upper limit of the assay range was >500-fold for all serotypes, and assay variability was <20% relative standard deviation (RSD) for all serotypes. IgG antibody measurements were shown to be serotype-specific (some cross-reactivity was observed only between the structurally related serotypes 6A and 6B as well as 19A and 19F), and no interference was observed between the serotypes when the assay was performed in the 13-plex format compared to the singleplex assays. The 13-plex dLIA platform developed by Pfizer Inc. generates up to 143 test results in a single 96-well plate and is a suitable replacement of the enzyme-linked immunosorbent assay (ELISA) platform for evaluating vaccine clinical trials

Group a streptococcal carriage and seroepidemiology in children up to 10 years of age in Australia

Background: Group A streptococci (GAS) and other β-hemolytic streptococci (BHS) cause pharyngitis, severe invasive disease and serious nonsuppurative sequelae including rheumatic heart disease and post streptococcal glomerulonephritis. The aim of this study was to assess carriage rates and anti-streptococcal C5a peptidase (anti-SCP) IgG levels and identify epidemiologic factors related to carriage or seropositivity in Australian children. Methods: A throat swab and blood sample were collected for microbiological and serological analysis (anti-SCP IgG) in 542 healthy children aged 0-10 years. Sequence analysis of the SCP gene was performed. Serological analysis used a competitive Luminex Immunoassay designed to preferentially detect functional antibody. Results: GAS-positive culture prevalence in throat swabs was 5.0% (range 0-10%), with the highest rate in 5 and 9 years old children. The rate of non-GAS BHS carriage was low