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I detta dokument beskrivs rekommenderade metoder för blododling inkluderat fungemi(svampinfektion i blod) samt metodik för diagnostik av endokardit och misstanke om infektion relaterad till kärlkatetrar. Rekommendationerna är baserade på internationella riktlinjer och relevant vetenskaplig litteratur. Där det finns icke tillräcklig vetenskapligt stöd har arbetsgruppen antingen avstått från en rekommendation eller givit en rekommendation baserad på de ingående medlemmarnas erfarenhet och kunnande.

BACKGROUND: The prevalence of fungal bloodstream infections (BSI), especially candidaemia, has been increasing globally during the last decades. Fungal diagnosis is still challenging due to the slow growth of fungal microorganisms and need for special expertise. Fungal polymicrobial infections further complicate the diagnosis and extend the time required. Epidemiological data are vital to generate effective empirical treatment strategies. OBJECTIVES: The overall aim of this project is to describe the epidemiology of monomicrobial candidaemia and polymicrobial BSI, both with mixed fungaemia and with mixed Candida/bacterial BSIs. METHODS: We conducted a single-centre retrospective epidemiological study that encompasses 950,161 blood cultures during the years 2010 to 2020. The epidemiology of monomicrobial and polymicrobial candidaemia episodes were investigated from the electronic records. RESULTS: We found that 1334 candidaemia episodes were identified belonging to 1144 individual patients during 2010 to 2020. Candida albicans was the most prevalent species detected in candidaemia patients, representing 57.7% of these episodes. Nakaseomyces (Candida) glabrata and Candida parapsilosis complex showed an increasing trend compared to previous studies, whereas Candida albicans demonstrated a decrease. 19.8% of these episodes were polymicrobial and 17% presented with mixed Candida/bacterial BSIs while 2.8% were mixed fungaemia. C. albicans and N. glabrata were the most common combination (51.4%) in mixed fungaemia episodes. Enterococcus and Lactobacillus spp. were the most common bacteria isolated in mixed Candida/bacterial BSIs. CONCLUSIONS: Polymicrobial growth with candidaemia is common, mostly being mixed Candida/bacterial BSIs. C. albicans was detected in more than half of all the candidaemia patients however showed a decreasing trend in time, whereas an increase is noteworthy in C. parapsilosis complex and N. glabrata.

UTI-lizer is a recent digital format for easy-to-use culture-based detection, preliminary identification, and quantification of bacteria in urine at the point of care (PoC). This study aimed to evaluate the diagnostic accuracy of UTI-lizer tests for detection of bacteriuria caused by five common bacterial species: Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, Proteus mirabilis, and Staphylococcus saprophyticus. We evaluated the accuracy of UTI-lizer tests by comparing test results of UTI-lizer with those of current standard bacterial culture-based diagnostics in clinical microbiology laboratories in a retrospective and a prospective study. Comparator methods were classical bacterial culture in combination with matrix-assisted laser desorption/ionization-time of flight mass spectrometry mediated bacterial identification. In the retrospective study, we tested 104 urine samples with in-panel microorganisms, plain urethral microbiota, and culture-negative samples. In the prospective study, we used 137 urine samples within 10 hours of their collection at general practitioner clinics. The retrospective study demonstrated 100% sensitivity and specificity in the detection of bacteriuria, and 98.6% sensitivity and 96.8% specificity in identifying primary pathogens with UTI-lizer when compared to clinical standards. S. saprophyticus and E. coli could not be distinguished. The combined nitrite and esterase test predicted the presence of bacteriuria in only 36.5% of cases. The prospective study demonstrated 100% sensitivity and 89.6% specificity in the detection of significant bacteriuria for in-panel microorganisms with a coverage rate of 88.3% (121/137). This study indicates that digital dipsticks are a promising alternative for the detection of the five main pathogens that cause the vast majority of urinary tract infections (UTIs). The results demonstrate that digital dipsticks have the potential to uniquely provide—in primary care or at the point of care—a, QC 20231218

The objectives of this study were to assess Candida spp. distribution and antifungal resistance of candidaemia across Europe. Isolates were collected as part of the third ECMM Candida European multicentre observational study, conducted from 01 to 07-07-2018 to 31-03-2022. Each centre (maximum number/country determined by population size) included ∼10 consecutive cases. Isolates were referred to central laboratories and identified by morphology and MALDI-TOF, supplemented by ITS-sequencing when needed. EUCAST MICs were determined for five antifungals. fks sequencing was performed for echinocandin resistant isolates. The 399 isolates from 41 centres in 17 countries included C. albicans (47.1%), C. glabrata (22.3%), C. parapsilosis (15.0%), C. tropicalis (6.3%), C. dubliniensis and C. krusei (2.3% each) and other species (4.8%). Austria had the highest C. albicans proportion (77%), Czech Republic, France and UK the highest C. glabrata proportions (25–33%) while Italy and Turkey had the highest C. parapsilosis proportions (24–26%). All isolates were amphotericin B susceptible. Fluconazole resistance was found in 4% C. tropicalis, 12% C. glabrata (from six countries across Europe), 17% C. parapsilosis (from Greece, Italy, and Turkey) and 20% other Candida spp. Four isolates were anidulafungin and micafungin resistant/non-wild-type and five resistant to micafungin only. Three/3 and 2/5 of these were sequenced and harboured fks-alterations including a novel L657W in C. parapsilosis. The epidemiology varied among centres and countries. Acquired echinocandin resistance was rare but included differential susceptibility to anidulafungin and micafungin, and resistant C. parapsilosis. Fluconazole and voriconazole cross-resistance was common in C. glabrata and C. parapsilosis but with different geographical prevalence., The objectives of this study were to assess Candida spp. distribution and antifungal resistance of candidaemia across Europe. Isolates were collected as part of the third ECMM Candida European multicentre observational study, conducted from 01 to 07-07-2018 to 31-03-2022. Each centre (maximum number/country determined by population size) included ∼10 consecutive cases. Isolates were referred to central laboratories and identified by morphology and MALDI-TOF, supplemented by ITS-sequencing when needed. EUCAST MICs were determined for five antifungals. fks sequencing was performed for echinocandin resistant isolates. The 399 isolates from 41 centres in 17 countries included C. albicans (47.1%), C. glabrata (22.3%), C. parapsilosis (15.0%), C. tropicalis (6.3%), C. dubliniensis and C. krusei (2.3% each) and other species (4.8%). Austria had the highest C. albicans proportion (77%), Czech Republic, France and UK the highest C. glabrata proportions (25–33%) while Italy and Turkey had the highest C. parapsilosis proportions (24–26%). All isolates were amphotericin B susceptible. Fluconazole resistance was found in 4% C. tropicalis, 12% C. glabrata (from six countries across Europe), 17% C. parapsilosis (from Greece, Italy, and Turkey) and 20% other Candida spp. Four isolates were anidulafungin and micafungin resistant/non-wild-type and five resistant to micafungin only. Three/3 and 2/5 of these were sequenced and harboured fks-alterations including a novel L657W in C. parapsilosis. The epidemiology varied among centres and countries. Acquired echinocandin resistance was rare but included differential susceptibility to anidulafungin and micafungin, and resistant C. parapsilosis. Fluconazole and voriconazole cross-resistance was common in C. glabrata and C. parapsilosis but with different geographical prevalence.

[Background] The European Confederation of Medical Mycology (ECMM) collected data on epidemiology, risk factors, treatment, and outcomes of patients with culture-proven candidaemia across Europe to assess how adherence to guideline recommendations is associated with outcomes., [Methods] In this observational cohort study, 64 participating hospitals located in 20 European countries, with the number of eligible hospitals per country determined by population size, included the first ten consecutive adults with culture-proven candidaemia after July 1, 2018, and entered data into the ECMM Candida Registry (FungiScope CandiReg). We assessed ECMM Quality of Clinical Candidaemia Management (EQUAL Candida) scores reflecting adherence to recommendations of the European Society of Clinical Microbiology and Infectious Diseases and the Infectious Diseases Society of America guidelines., [Findings] 632 patients with candidaemia were included from 64 institutions. Overall 90-day mortality was 43% (265/617), and increasing age, intensive care unit admission, point increases in the Charlson comorbidity index score, and Candida tropicalis as causative pathogen were independent baseline predictors of mortality in Cox regression analysis. EQUAL Candida score remained an independent predictor of mortality in the multivariable Cox regression analyses after adjusting for the baseline predictors, even after restricting the analysis to patients who survived for more than 7 days after diagnosis (adjusted hazard ratio 1·08 [95% CI 1·04–1·11; p<0·0001] in patients with a central venous catheter and 1·09 [1·05–1·13; p<0·0001] in those without one, per one score point decrease). Median duration of hospital stay was 15 days (IQR 4–30) after diagnosis of candidaemia and was extended specifically for completion of parenteral therapy in 100 (16%) of 621 patients. Initial echinocandin treatment was associated with lower overall mortality and longer duration of hospital stay among survivors than treatment with other antifungals., [Interpretation] Although overall mortality in patients with candidaemia was high, our study indicates that adherence to clinical guideline recommendations, reflected by higher EQUAL Candida scores, might increase survival. New antifungals, with similar activity as current echinocandins but with longer half-lives or oral bioavailability, are needed to reduce duration of hospital stay.

Background: The European Confederation of Medical Mycology (ECMM) collected data on epidemiology, risk factors, treatment, and outcomes of patients with culture-proven candidaemia across Europe to assess how adherence to guideline recommendations is associated with outcomes. Methods: In this observational cohort study, 64 participating hospitals located in 20 European countries, with the number of eligible hospitals per country determined by population size, included the first ten consecutive adults with culture-proven candidaemia after July 1, 2018, and entered data into the ECMM Candida Registry (FungiScope CandiReg). We assessed ECMM Quality of Clinical Candidaemia Management (EQUAL Candida) scores reflecting adherence to recommendations of the European Society of Clinical Microbiology and Infectious Diseases and the Infectious Diseases Society of America guidelines. Findings: 632 patients with candidaemia were included from 64 institutions. Overall 90-day mortality was 43% (265/617), and increasing age, intensive care unit admission, point increases in the Charlson comorbidity index score, and Candida tropicalis as causative pathogen were independent baseline predictors of mortality in Cox regression analysis. EQUAL Candida score remained an independent predictor of mortality in the multivariable Cox regression analyses after adjusting for the baseline predictors, even after restricting the analysis to patients who survived for more than 7 days after diagnosis (adjusted hazard ratio 1·08 [95% CI 1·04–1·11; p<0·0001] in patients with a central venous catheter and 1·09 [1·05–1·13; p<0·0001] in those without one, per one score point decrease). Median duration of hospital stay was 15 days (IQR 4–30) after diagnosis of candidaemia and was extended specifically for completion of parenteral therapy in 100 (16%) of 621 patients. Initial echinocandin treatment was associated with lower overall mortality and longer duration of hospital stay among survivors than t

The objectives of this study were to assess Candida spp. distribution and antifungal resistance of candidaemia across Europe. Isolates were collected as part of the third ECMM Candida European multicentre observational study, conducted from 01 to 07-07-2018 to 31-03-2022. Each centre (maximum number/country determined by population size) included ∼10 consecutive cases. Isolates were referred to central laboratories and identified by morphology and MALDI-TOF, supplemented by ITS-sequencing when needed. EUCAST MICs were determined for five antifungals. fks sequencing was performed for echinocandin resistant isolates. The 399 isolates from 41 centres in 17 countries included C. albicans (47.1%), C. glabrata (22.3%), C. parapsilosis (15.0%), C. tropicalis (6.3%), C. dubliniensis and C. krusei (2.3% each) and other species (4.8%). Austria had the highest C. albicans proportion (77%), Czech Republic, France and UK the highest C. glabrata proportions (25–33%) while Italy and Turkey had the highest C. parapsilosis proportions (24–26%). All isolates were amphotericin B susceptible. Fluconazole resistance was found in 4% C. tropicalis, 12% C. glabrata (from six countries across Europe), 17% C. parapsilosis (from Greece, Italy, and Turkey) and 20% other Candida spp. Four isolates were anidulafungin and micafungin resistant/non-wild-type and five resistant to micafungin only. Three/3 and 2/5 of these were sequenced and harboured fks-alterations including a novel L657W in C. parapsilosis. The epidemiology varied among centres and countries. Acquired echinocandin resistance was rare but included differential susceptibility to anidulafungin and micafungin, and resistant C. parapsilosis. Fluconazole and voriconazole cross-resistance was common in C. glabrata and C. parapsilosis but with different geographical prevalence., The objectives of this study were to assess Candida spp. distribution and antifungal resistance of candidaemia across Europe. Isolates were collected as part of the third ECMM Candida European multicentre observational study, conducted from 01 to 07-07-2018 to 31-03-2022. Each centre (maximum number/country determined by population size) included ∼10 consecutive cases. Isolates were referred to central laboratories and identified by morphology and MALDI-TOF, supplemented by ITS-sequencing when needed. EUCAST MICs were determined for five antifungals. fks sequencing was performed for echinocandin resistant isolates. The 399 isolates from 41 centres in 17 countries included C. albicans (47.1%), C. glabrata (22.3%), C. parapsilosis (15.0%), C. tropicalis (6.3%), C. dubliniensis and C. krusei (2.3% each) and other species (4.8%). Austria had the highest C. albicans proportion (77%), Czech Republic, France and UK the highest C. glabrata proportions (25–33%) while Italy and Turkey had the highest C. parapsilosis proportions (24–26%). All isolates were amphotericin B susceptible. Fluconazole resistance was found in 4% C. tropicalis, 12% C. glabrata (from six countries across Europe), 17% C. parapsilosis (from Greece, Italy, and Turkey) and 20% other Candida spp. Four isolates were anidulafungin and micafungin resistant/non-wild-type and five resistant to micafungin only. Three/3 and 2/5 of these were sequenced and harboured fks-alterations including a novel L657W in C. parapsilosis. The epidemiology varied among centres and countries. Acquired echinocandin resistance was rare but included differential susceptibility to anidulafungin and micafungin, and resistant C. parapsilosis. Fluconazole and voriconazole cross-resistance was common in C. glabrata and C. parapsilosis but with different geographical prevalence.

The T2Bacteria panel is a direct-from-blood assay that delivers rapid results, targeting E. coli, S. aureus, K. pneumoniae, A. baumanii, P. aeruginosa, and E. faecium (ESKAPE pathogens). In this study, T2Bacteria and T2Candida (targeting C. albicans/C. tropicalis, C. glabrata/C. krusei, and C. parapsilosis) were evaluated in parallel with blood cultures in 101 consecutive surgical patients with suspected intra-abdominal infection admitted to the intensive care unit or high dependency unit. Fifteen patients had bacteremia, with T2Bacteria correctly identifying all on-panel (n = 8) pathogens. T2Bacteria was positive in 19 additional patients, 11 of whom had supportive cultures from other normally sterile sites (newly inserted drains, perioperative cultures or blood cultures) within seven days. Six of these eleven patients (55%) received broad-spectrum antibiotics at the sampling time. T2Candida identified the two cases of blood-culture-positive candidemia and was positive in seven additional patients, three of whom were confirmed to have intra-abdominal candidiasis. Of four patients with concurrent T2Bacteria and T2Candida positivity, only one patient had positive blood cultures (candidemia), while three out of four patients had supporting microbiological evidence of a mixed infection. T2Bacteria and T2Candida were fast and accurate in diagnosing on-panel bloodstream infections, and T2Bacteria was able to detect culture-negative intra-abdominal infections.

The T2Bacteria panel is a direct-from-blood assay that delivers rapid results, targeting E. coli, S. aureus, K. pneumoniae, A. baumanii, P. aeruginosa, and E. faecium (ESKAPE pathogens). In this study, T2Bacteria and T2Candida (targeting C. albicans/C. tropicalis, C. glabrata/C. krusei, and C. parapsilosis) were evaluated in parallel with blood cultures in 101 consecutive surgical patients with suspected intra-abdominal infection admitted to the intensive care unit or high dependency unit. Fifteen patients had bacteremia, with T2Bacteria correctly identifying all on-panel (n = 8) pathogens. T2Bacteria was positive in 19 additional patients, 11 of whom had supportive cultures from other normally sterile sites (newly inserted drains, perioperative cultures or blood cultures) within seven days. Six of these eleven patients (55%) received broad-spectrum antibiotics at the sampling time. T2Candida identified the two cases of blood-culture-positive candidemia and was positive in seven additional patients, three of whom were confirmed to have intra-abdominal candidiasis. Of four patients with concurrent T2Bacteria and T2Candida positivity, only one patient had positive blood cultures (candidemia), while three out of four patients had supporting microbiological evidence of a mixed infection. T2Bacteria and T2Candida were fast and accurate in diagnosing on-panel bloodstream infections, and T2Bacteria was able to detect culture-negative intra-abdominal infections.

The T2Bacteria panel is a direct-from-blood assay that delivers rapid results, targeting E. coli, S. aureus, K. pneumoniae, A. baumanii, P. aeruginosa, and E. faecium (ESKAPE pathogens). In this study, T2Bacteria and T2Candida (targeting C. albicans/C. tropicalis, C. glabrata/C. krusei, and C. parapsilosis) were evaluated in parallel with blood cultures in 101 consecutive surgical patients with suspected intra-abdominal infection admitted to the intensive care unit or high dependency unit. Fifteen patients had bacteremia, with T2Bacteria correctly identifying all on-panel (n = 8) pathogens. T2Bacteria was positive in 19 additional patients, 11 of whom had supportive cultures from other normally sterile sites (newly inserted drains, perioperative cultures or blood cultures) within seven days. Six of these eleven patients (55%) received broad-spectrum antibiotics at the sampling time. T2Candida identified the two cases of blood-culture-positive candidemia and was positive in seven additional patients, three of whom were confirmed to have intra-abdominal candidiasis. Of four patients with concurrent T2Bacteria and T2Candida positivity, only one patient had positive blood cultures (candidemia), while three out of four patients had supporting microbiological evidence of a mixed infection. T2Bacteria and T2Candida were fast and accurate in diagnosing on-panel bloodstream infections, and T2Bacteria was able to detect culture-negative intra-abdominal infections.

The T2Bacteria panel is a direct-from-blood assay that delivers rapid results, targeting E. coli, S. aureus, K. pneumoniae, A. baumanii, P. aeruginosa, and E. faecium (ESKAPE pathogens). In this study, T2Bacteria and T2Candida (targeting C. albicans/C. tropicalis, C. glabrata/C. krusei, and C. parapsilosis) were evaluated in parallel with blood cultures in 101 consecutive surgical patients with suspected intra-abdominal infection admitted to the intensive care unit or high dependency unit. Fifteen patients had bacteremia, with T2Bacteria correctly identifying all on-panel (n = 8) pathogens. T2Bacteria was positive in 19 additional patients, 11 of whom had supportive cultures from other normally sterile sites (newly inserted drains, perioperative cultures or blood cultures) within seven days. Six of these eleven patients (55%) received broad-spectrum antibiotics at the sampling time. T2Candida identified the two cases of blood-culture-positive candidemia and was positive in seven additional patients, three of whom were confirmed to have intra-abdominal candidiasis. Of four patients with concurrent T2Bacteria and T2Candida positivity, only one patient had positive blood cultures (candidemia), while three out of four patients had supporting microbiological evidence of a mixed infection. T2Bacteria and T2Candida were fast and accurate in diagnosing on-panel bloodstream infections, and T2Bacteria was able to detect culture-negative intra-abdominal infections.

The T2Bacteria panel is a direct-from-blood assay that delivers rapid results, targeting E. coli, S. aureus, K. pneumoniae, A. baumanii, P. aeruginosa, and E. faecium (ESKAPE pathogens). In this study, T2Bacteria and T2Candida (targeting C. albicans/C. tropicalis, C. glabrata/C. krusei, and C. parapsilosis) were evaluated in parallel with blood cultures in 101 consecutive surgical patients with suspected intra-abdominal infection admitted to the intensive care unit or high dependency unit. Fifteen patients had bacteremia, with T2Bacteria correctly identifying all on-panel (n = 8) pathogens. T2Bacteria was positive in 19 additional patients, 11 of whom had supportive cultures from other normally sterile sites (newly inserted drains, perioperative cultures or blood cultures) within seven days. Six of these eleven patients (55%) received broad-spectrum antibiotics at the sampling time. T2Candida identified the two cases of blood-culture-positive candidemia and was positive in seven additional patients, three of whom were confirmed to have intra-abdominal candidiasis. Of four patients with concurrent T2Bacteria and T2Candida positivity, only one patient had positive blood cultures (candidemia), while three out of four patients had supporting microbiological evidence of a mixed infection. T2Bacteria and T2Candida were fast and accurate in diagnosing on-panel bloodstream infections, and T2Bacteria was able to detect culture-negative intra-abdominal infections.

Objectives The performance of blood cultures (BC) relies on optimal sampling. Sepsis guidelines do not specify which sampling protocol to use, but recommend two sets of BC bottles, each set containing one aerobic and one anaerobic bottle. For the single-site sampling (SSS) protocol, only one venipuncture is performed for all four bottles. The predominating multi-site sampling (MSS) protocol implies that BC bottles are collected from two separate venipuncture sites. The aim of this study was to compare SSS and MSS. Primary outcomes were number of BC sets collected, sample volume and diagnostic performance. Methods This was a retrospective clinical study comparing BC results in an emergency department before and after changing the sampling protocol to SSS from MSS. All BC samples were incubated in the BacT/ALERT BC system. Results The analysis included 5,248 patients before and 5,364 patients after the implementation of SSS. There was a significantly higher proportion of positive BCs sampled with SSS compared to MSS, 1,049/5,364 (19.56%) and 932/5,248 (17.76%) respectively (P=0.018). This difference was due to a higher proportion of solitary BC sets (two BC bottles) in MSS. Analyzing only patients with the recommended four BC bottles, there was no difference in positivity. SSS had a higher proportion of BC bottles with the recommended sample volumes of 8-12 ml than MSS (P<0.001). Conclusions Changing the sampling protocol to SSS from MSS resulted in higher positivity rates, higher sample volume and fewer solitary BC sets. These advantages of SSS should be considered in future sepsis guidelines., Correction for Ekwall-Larson et al., “Single-Site Sampling versus Multisite Sampling for Blood Cultures: a Retrospective Clinical Study”. Journal of Clinical Microbiology, vol. 60(9), e01180-22. DOI 10.1128/jcm.01180-22

Emerging research suggests gut microbiome may play a role in pancreatic cancer initiation and progression, but cultivation of the cancer microbiome remains challenging. This pilot study aims to investigate the possibility to cultivate pancreatic microbiome from pancreatic cystic lesions associated with invasive cancer. Intra-operatively acquired pancreatic cyst fluid samples showed culture-positivity mainly in the intraductal papillary mucinous neoplasm (IPMN) group of lesions. MALDI-TOF MS profiling analysis shows Gammaproteobacteria and Bacilli dominate among individual bacteria isolates. Among cultivated bacteria, Gammaproteobacteria, particularly Klebsiella pneumoniae, but also Granulicatella adiacens and Enterococcus faecalis, demonstrate consistent pathogenic properties in pancreatic cell lines tested in ex vivo co-culture models. Pathogenic properties include intracellular survival capability, cell death induction, or causing DNA double-strand breaks in the surviving cells resembling genotoxic effects. This study provides new insights into the role of the pancreatic microbiota in the intriguing link between pancreatic cystic lesions and cancer.

Emerging research suggests gut microbiome may play a role in pancreatic cancer initiation and progression, but cultivation of the cancer microbiome remains challenging. This pilot study aims to investigate the possibility to cultivate pancreatic microbiome from pancreatic cystic lesions associated with invasive cancer. Intra-operatively acquired pancreatic cyst fluid samples showed culture-positivity mainly in the intraductal papillary mucinous neoplasm (IPMN) group of lesions. MALDI-TOF MS profiling analysis shows Gammaproteobacteria and Bacilli dominate among individual bacteria isolates. Among cultivated bacteria, Gammaproteobacteria, particularly Klebsiella pneumoniae, but also Granulicatella adiacens and Enterococcus faecalis, demonstrate consistent pathogenic properties in pancreatic cell lines tested in ex vivo co-culture models. Pathogenic properties include intracellular survival capability, cell death induction, or causing DNA double-strand breaks in the surviving cells resembling genotoxic effects. This study provides new insights into the role of the pancreatic microbiota in the intriguing link between pancreatic cystic lesions and cancer.

Emerging research suggests gut microbiome may play a role in pancreatic cancer initiation and progression, but cultivation of the cancer microbiome remains challenging. This pilot study aims to investigate the possibility to cultivate pancreatic microbiome from pancreatic cystic lesions associated with invasive cancer. Intra-operatively acquired pancreatic cyst fluid samples showed culture-positivity mainly in the intraductal papillary mucinous neoplasm (IPMN) group of lesions. MALDI-TOF MS profiling analysis shows Gammaproteobacteria and Bacilli dominate among individual bacteria isolates. Among cultivated bacteria, Gammaproteobacteria, particularly Klebsiella pneumoniae, but also Granulicatella adiacens and Enterococcus faecalis, demonstrate consistent pathogenic properties in pancreatic cell lines tested in ex vivo co-culture models. Pathogenic properties include intracellular survival capability, cell death induction, or causing DNA double-strand breaks in the surviving cells resembling genotoxic effects. This study provides new insights into the role of the pancreatic microbiota in the intriguing link between pancreatic cystic lesions and cancer.

The human biliary system, a mucosal barrier tissue connecting the liver and intestine, is an organ often affected by serious inflammatory and malignant diseases. Although these diseases are linked to immunological processes, the biliary system represents an unexplored immunological niche. By combining endoscopy-guided sampling of the biliary tree with a high-dimensional analysis approach, comprehensive mapping of the human biliary immunological landscape in patients with primary sclerosing cholangitis (PSC), a severe biliary inflammatory disease, was conducted. Major differences in immune cell composition in bile ducts compared to blood were revealed. Furthermore, biliary inflammation in patients with PSC was characterized by high presence of neutrophils and T cells as compared to control individuals without PSC. The biliary T cells displayed a CD103+CD69+ effector memory phenotype, a combined gut and liver homing profile, and produced interleukin-17 (IL-17) and IL-22. Biliary neutrophil infiltration in PSC associated with CXCL8, possibly produced by resident T cells, and CXCL16 was linked to the enrichment of T cells. This study uncovers the immunological niche of human bile ducts, defines a local immune network between neutrophils and biliary-resident T cells in PSC, and provides a resource for future studies of the immune responses in biliary disorders.

Emerging research suggests gut microbiome may play a role in pancreatic cancer initiation and progression, but cultivation of the cancer microbiome remains challenging. This pilot study aims to investigate the possibility to cultivate pancreatic microbiome from pancreatic cystic lesions associated with invasive cancer. Intra-operatively acquired pancreatic cyst fluid samples showed culture-positivity mainly in the intraductal papillary mucinous neoplasm (IPMN) group of lesions. MALDI-TOF MS profiling analysis shows Gammaproteobacteria and Bacilli dominate among individual bacteria isolates. Among cultivated bacteria, Gammaproteobacteria, particularly Klebsiella pneumoniae, but also Granulicatella adiacens and Enterococcus faecalis, demonstrate consistent pathogenic properties in pancreatic cell lines tested in ex vivo co-culture models. Pathogenic properties include intracellular survival capability, cell death induction, or causing DNA double-strand breaks in the surviving cells resembling genotoxic effects. This study provides new insights into the role of the pancreatic microbiota in the intriguing link between pancreatic cystic lesions and cancer.

Emerging research suggests gut microbiome may play a role in pancreatic cancer initiation and progression, but cultivation of the cancer microbiome remains challenging. This pilot study aims to investigate the possibility to cultivate pancreatic microbiome from pancreatic cystic lesions associated with invasive cancer. Intra-operatively acquired pancreatic cyst fluid samples showed culture-positivity mainly in the intraductal papillary mucinous neoplasm (IPMN) group of lesions. MALDI-TOF MS profiling analysis shows Gammaproteobacteria and Bacilli dominate among individual bacteria isolates. Among cultivated bacteria, Gammaproteobacteria, particularly Klebsiella pneumoniae, but also Granulicatella adiacens and Enterococcus faecalis, demonstrate consistent pathogenic properties in pancreatic cell lines tested in ex vivo co-culture models. Pathogenic properties include intracellular survival capability, cell death induction, or causing DNA double-strand breaks in the surviving cells resembling genotoxic effects. This study provides new insights into the role of the pancreatic microbiota in the intriguing link between pancreatic cystic lesions and cancer.

Streptococcus pneumoniae urinary antigen tests (UATs) may be interpreted using automatic readers to potentially automate sample incubation and provide standardized results reading. Here, we evaluated four UATs the BinaxNOW S. pneumoniae Antigen Card (Abbott, Chicago, IL, USA), ImmuView S. pneumoniae and Legionella (SSI Diagnostica, Hillerød, Denmark), STANDARD F S. pneumoniae Ag FIA (SD Biosensor, Gyeonggi, South Korea), and Sofia S. pneumoniae FIA (Quidel Corporation, San Diego, CA, USA) with their respective benchtop readers for their ability to detect S. pneumoniae urinary antigen. We found that these assays had a sensitivity of 76.9–86.5%, and specificity of 84.2–89.7%, with no significant difference found among the four UATs. The assays had a high level of agreement with each other, with 84.5% of samples testing consistently across all four assays. The automatically and visually read test results from the two immunochromatographic assays, BinaxNOW and ImmuView, were compared and showed excellent agreement between the two types of reading. Immunofluorescent-based assays, Sofia and STANDARD F, had significantly less time to detect compared to the two immunochromatographic assays due to having less assay setup procedures and shorter sample incubation times. In conclusion, the four UATs performed similarly in the detection of S. pneumoniae urinary antigen, and readers can bring increased flexibility to running UATs in the clinical routine.

Urinary antigen tests (UATs) are often used to diagnose Legionnaires' disease as they are rapid and easy to perform on readily obtainable urine samples without the need for specialized skills compared to conventional methods. Recently developed automated readers for UATs may provide objective results interpretation, especially in cases of weak result bands. Using 53 defined patient urine samples, we evaluated the performance of the BinaxNOW Legionella Antigen Card (Abbott), ImmuView S. pneumoniae and Legionella (SSI Diagnostica), STANDARD F Legionella Ag FIA (SD Biosensor), and Sofia Legionella FIA (Quidel) simultaneously with their respective automated readers. Automatic and visual interpretation of result bands were also compared for the immunochromatography-based BinaxNOW and ImmuView UATs. Overall sensitivity and specificity of Legionella UATs were 53.9-61.5% and 90.0-94.9%, respectively. All four UATs successfully detected all samples from L. pneumophila serogroup 1-positive patients, but most failed to detect samples for Legionella spp., or other serogroups. Automatic results interpretation of results was found to be mostly concordant with visual results reading. In conclusion, the performance of the four UATs were similar to each other in the detection of Legionella urinary antigen with no major difference between automated or visual results reading.

Blood culture (BC) often fails to detect bloodstream microorganisms in sepsis. However, molecular diagnostics hold great potential. The molecular method PCR/electrospray ionization-mass spectrometry (PCR/ESI-MS) can detect DNA from hundreds of different microorganisms in whole blood. The aim of the present study was to evaluate the performance of this method in a multicenter study including 16 teaching hospitals in the USA (n=13) and Europe (n=3). First, on 2,754 contrived whole blood samples, with or without spiked microorganisms, PCR/ESI-MS produced 99.1% true positive and 97.2% true negative results. Secondly, among 1,460 patients with suspected sepsis (sepsis-2 definition), BC and PCR/ESI-MS on whole blood were positive in 14.6% and 25.6% of cases, respectively, with the following result combinations: BC+/PCR/ESI-MS-, 4.3%; BC+/PCR/ESI-MS+, 10.3%; BC-/PCR/ESI-MS+, 15.3%; and BC-/PCR/ESI-MS-, 70.1%. Compared with BC, PCR/ESI-MS showed the following sensitivities (coagulase-negative staphylococci not included): Gram-positive bacteria, 58%; Gram-negative bacteria, 78%; and Candida species, 83%. The specificities were > 94% for all individual species. Patients treated with prior antimicrobial medications (n=603) had significantly increased PCR/ESI-MS positivity rates compared with patients without prior antimicrobial treatment, 31% vs 22% (p<0.0001), with pronounced differences for Gram-negative bacteria and Candida species. In conclusion, PCR/ESI-MS showed excellent performance on contrived samples. On clinical samples, it showed high specificities, moderately high sensitivities for Gram-negative bacteria and Candida species, and elevated positivity rates during antimicrobial treatment. These promising results encourage further development of molecular diagnostics on whole blood for detection of bloodstream microorganisms in sepsis., Funding Agencies:Ibis Biosciences, a division of Abbott Abbott LaboratoriesCepheid ContraFect Nabriva Therapeutics AG Lajolla Pharmaceuticals Alere Corporation Spectral Diagnostics Ferring Pharmaceuticals Inflammix VICTASm Sanofi Pasteur Inc. Curetis GmbH PfizerMerck & CompanyCidara Therapeutics Inc. Shire Aridis Pharmaceuticals Inc. Epigenomics Inc. Roche Holding GenentechFinch Therapeutics Seres Therapeutics Inc. Diasorin Molecular Janssen Research and Development NeuMoDx Molecular Iterum Therapeutics International

Blood volume is the most important variable, for detection of microorganisms in blood cultures (BC). Most standards recommend 40-60 ml blood, collected in several BC bottles filled up to 10 ml. We measured blood volume in individual BC bottles, and analysed the association of hospital, bottle type, day of the week, daily sampling time, and age and gender of the patient, with sampling volume and BC result. The variation in blood volume per BC bottle was analysed in a mixed linear model using hospital, bottle type, weekday, sampling time, age and gender as fixed factors, and patient ID and episode as random factors to control for repetitive sampling of individual patients. Only 18 % of all bottles were filled with the recommended 8-10 ml, and 47 % were filled with less than 8 ml. The mean (±SE) volume was larger in positive 9.09 (±0.15) compared to negative bottles 8.47 (±0.07) (p<0.001). Blood volume was larger in BacT/ALERT-FA Plus than in -FN Plus BC bottles (p<0.001). There was significantly lower volumes collected during the night (p<0.001). The volume of blood collected decreased significantly with increasing patient age (p<0.001). Larger volumes were collected from males compared to females, 8.78 (±0.06) vs. 8.36 (±0.06) ml (mean ± SE) respectively (p<0.001). The odds of detecting a positive patient increases with 13 % for each additional ml blood drawn. Our results show that we need to work actively with development of blood sampling routines to overcome age and gender effects, and to optimize blood sampling volumes.

Blood volume is the most important variable, for detection of microorganisms in blood cultures (BC). Most standards recommend 40-60 ml blood, collected in several BC bottles filled up to 10 ml. We measured blood volume in individual BC bottles, and analysed the association of hospital, bottle type, day of the week, daily sampling time, and age and gender of the patient, with sampling volume and BC result. The variation in blood volume per BC bottle was analysed in a mixed linear model using hospital, bottle type, weekday, sampling time, age and gender as fixed factors, and patient ID and episode as random factors to control for repetitive sampling of individual patients. Only 18 % of all bottles were filled with the recommended 8-10 ml, and 47 % were filled with less than 8 ml. The mean (±SE) volume was larger in positive 9.09 (±0.15) compared to negative bottles 8.47 (±0.07) (p<0.001). Blood volume was larger in BacT/ALERT-FA Plus than in -FN Plus BC bottles (p<0.001). There was significantly lower volumes collected during the night (p<0.001). The volume of blood collected decreased significantly with increasing patient age (p<0.001). Larger volumes were collected from males compared to females, 8.78 (±0.06) vs. 8.36 (±0.06) ml (mean ± SE) respectively (p<0.001). The odds of detecting a positive patient increases with 13 % for each additional ml blood drawn. Our results show that we need to work actively with development of blood sampling routines to overcome age and gender effects, and to optimize blood sampling volumes.

The usefulness of pneumococcal urinary antigen tests (UATs) in severe pneumococcal infection relies heavily on the performance in bacteremic patients. Fluorescence technology and automatic reading of test results may improve UAT performance. We evaluated the automatically read Sofia S. pneumoniae FIA for diagnosing pneumococcal bloodstream infection (BSI) in hospitalized adult patients. First, the Sofia FIA was evaluated on 97 patients with pneumococcal (n = 47) and nonpneumococcal (n = 50) BSI and compared with results by the visually read BinaxNOW S. pneumoniae immunochromatographic test (ICT) and ImmuView S. pneumoniae and Legionella pneumophila ICT. In four cases (4.1%), the Sofia FIA showed invalid test results, three of which showed invalid results by the ImmuView ICT previously. Based on 93 valid cases, the Sofia FIA showed similar sensitivity (for both comparisons: 68% versus 62%; P = 0.45) and specificity (for both comparisons: 91% versus 93%; P = 1.00) as the visually read UATs. Second, the Sofia FIA was prospectively evaluated on 82 consecutive nonfrozen urine samples, detecting pneumococcal antigen in 10 of 14 (sensitivity, 71%) pneumococcal BSI patients, similarly to the visually and automatically read BinaxNOW ICT (both 12 of 14; sensitivity, 86%; P = 0.50). Of five nonpneumococcal BSI cases, the Sofia FIA showed an invalid test result in one case, but no positive UAT results were obtained. Thus, the sensitivity and specificity of the Sofia FIA were similar to the performance rates of other UATs in patients with BSI, but invalid test results are of concern for the usefulness in pneumococcal BSI., Funding Agency:Region Örebro County Council Research Committee OLL-723721

Meeting Abstract: PP4.109

Immunological mechanisms play a central role in the pathogenesis sclerosis (MS). In this study two groups of related immune variables are cytokines and metalloproteinases (MMPs), which regulate each other's production and activation. Cytokines play key role in pathogenesis and treatments of Cytokines are regulatory proteins secreted by a variety of cells. The pleiotropic action of cytokines include numerous effects on cells of the immune modulating immune responses. Cytokine receptors are crucial for the net effect of cytokines. The balance between Th1 and Th2 type cytokines might determine whether the immune response in MS is detrimental or beneficial. MMPs are a group of endopeptidases that degrade extracellular pro mechanisms in the genesis of inflammatory demyelination, such as recruitment, blood-brain barrier breakdown and myelin destruction are con be MMP-dependent processes. Aims of the study: 1. To investigate the levels of cytokine secreting blood and cerebrospinal fluid mononuclear cells (MNC) in patients with MS and controls; 2. To define further the cytokine disbalance in MS by examining the levels of cytokine receptors that are important in the net effect of cytokine function; 3. To study the levels of blood and CSF MNC in MS expressing mRNA of MMPs and their inhibitors; and 4. To examine the effects of IFN-[beta] on cytokines and MMPs in MS. Results: MS is associated with increased numbers of IL-6 and TNF-[alpha] secreting blood MNC and decreased numbers of IL-10 secreting cells compared to healthy subjects. In CSF, similar numbers of IL-6 and IL-10 secreting cells were observed in patients with MS and other neurological diseases (OND). Patients with MS also had higher levels of MMP-3, MMP-9 and tissue inhibitor of metalloproteinases-1 (TIMP-1) mRNA expressing blood MNC compared to controls. When patients with MS examined before vs. during 1 year of treatment with IFN-[beta], treatment resulted in decreased numbers of IL-6 and TNF-[alpha] secreting MNC

Immunological mechanisms play a central role in the pathogenesis sclerosis (MS). In this study two groups of related immune variables are cytokines and metalloproteinases (MMPs), which regulate each other's production and activation. Cytokines play key role in pathogenesis and treatments of Cytokines are regulatory proteins secreted by a variety of cells. The pleiotropic action of cytokines include numerous effects on cells of the immune modulating immune responses. Cytokine receptors are crucial for the net effect of cytokines. The balance between Th1 and Th2 type cytokines might determine whether the immune response in MS is detrimental or beneficial. MMPs are a group of endopeptidases that degrade extracellular pro mechanisms in the genesis of inflammatory demyelination, such as recruitment, blood-brain barrier breakdown and myelin destruction are con be MMP-dependent processes. Aims of the study: 1. To investigate the levels of cytokine secreting blood and cerebrospinal fluid mononuclear cells (MNC) in patients with MS and controls; 2. To define further the cytokine disbalance in MS by examining the levels of cytokine receptors that are important in the net effect of cytokine function; 3. To study the levels of blood and CSF MNC in MS expressing mRNA of MMPs and their inhibitors; and 4. To examine the effects of IFN-[beta] on cytokines and MMPs in MS. Results: MS is associated with increased numbers of IL-6 and TNF-[alpha] secreting blood MNC and decreased numbers of IL-10 secreting cells compared to healthy subjects. In CSF, similar numbers of IL-6 and IL-10 secreting cells were observed in patients with MS and other neurological diseases (OND). Patients with MS also had higher levels of MMP-3, MMP-9 and tissue inhibitor of metalloproteinases-1 (TIMP-1) mRNA expressing blood MNC compared to controls. When patients with MS examined before vs. during 1 year of treatment with IFN-[beta], treatment resulted in decreased numbers of IL-6 and TNF-[alpha] secreting MNC

Immunological mechanisms play a central role in the pathogenesis sclerosis (MS). In this study two groups of related immune variables are cytokines and metalloproteinases (MMPs), which regulate each other's production and activation. Cytokines play key role in pathogenesis and treatments of Cytokines are regulatory proteins secreted by a variety of cells. The pleiotropic action of cytokines include numerous effects on cells of the immune modulating immune responses. Cytokine receptors are crucial for the net effect of cytokines. The balance between Th1 and Th2 type cytokines might determine whether the immune response in MS is detrimental or beneficial. MMPs are a group of endopeptidases that degrade extracellular pro mechanisms in the genesis of inflammatory demyelination, such as recruitment, blood-brain barrier breakdown and myelin destruction are con be MMP-dependent processes. Aims of the study: 1. To investigate the levels of cytokine secreting blood and cerebrospinal fluid mononuclear cells (MNC) in patients with MS and controls; 2. To define further the cytokine disbalance in MS by examining the levels of cytokine receptors that are important in the net effect of cytokine function; 3. To study the levels of blood and CSF MNC in MS expressing mRNA of MMPs and their inhibitors; and 4. To examine the effects of IFN-[beta] on cytokines and MMPs in MS. Results: MS is associated with increased numbers of IL-6 and TNF-[alpha] secreting blood MNC and decreased numbers of IL-10 secreting cells compared to healthy subjects. In CSF, similar numbers of IL-6 and IL-10 secreting cells were observed in patients with MS and other neurological diseases (OND). Patients with MS also had higher levels of MMP-3, MMP-9 and tissue inhibitor of metalloproteinases-1 (TIMP-1) mRNA expressing blood MNC compared to controls. When patients with MS examined before vs. during 1 year of treatment with IFN-[beta], treatment resulted in decreased numbers of IL-6 and TNF-[alpha] secreting MNC

Immunological mechanisms play a central role in the pathogenesis sclerosis (MS). In this study two groups of related immune variables are cytokines and metalloproteinases (MMPs), which regulate each other's production and activation. Cytokines play key role in pathogenesis and treatments of Cytokines are regulatory proteins secreted by a variety of cells. The pleiotropic action of cytokines include numerous effects on cells of the immune modulating immune responses. Cytokine receptors are crucial for the net effect of cytokines. The balance between Th1 and Th2 type cytokines might determine whether the immune response in MS is detrimental or beneficial. MMPs are a group of endopeptidases that degrade extracellular pro mechanisms in the genesis of inflammatory demyelination, such as recruitment, blood-brain barrier breakdown and myelin destruction are con be MMP-dependent processes. Aims of the study: 1. To investigate the levels of cytokine secreting blood and cerebrospinal fluid mononuclear cells (MNC) in patients with MS and controls; 2. To define further the cytokine disbalance in MS by examining the levels of cytokine receptors that are important in the net effect of cytokine function; 3. To study the levels of blood and CSF MNC in MS expressing mRNA of MMPs and their inhibitors; and 4. To examine the effects of IFN-[beta] on cytokines and MMPs in MS. Results: MS is associated with increased numbers of IL-6 and TNF-[alpha] secreting blood MNC and decreased numbers of IL-10 secreting cells compared to healthy subjects. In CSF, similar numbers of IL-6 and IL-10 secreting cells were observed in patients with MS and other neurological diseases (OND). Patients with MS also had higher levels of MMP-3, MMP-9 and tissue inhibitor of metalloproteinases-1 (TIMP-1) mRNA expressing blood MNC compared to controls. When patients with MS examined before vs. during 1 year of treatment with IFN-[beta], treatment resulted in decreased numbers of IL-6 and TNF-[alpha] secreting MNC

Immunological mechanisms play a central role in the pathogenesis sclerosis (MS). In this study two groups of related immune variables are cytokines and metalloproteinases (MMPs), which regulate each other's production and activation. Cytokines play key role in pathogenesis and treatments of Cytokines are regulatory proteins secreted by a variety of cells. The pleiotropic action of cytokines include numerous effects on cells of the immune modulating immune responses. Cytokine receptors are crucial for the net effect of cytokines. The balance between Th1 and Th2 type cytokines might determine whether the immune response in MS is detrimental or beneficial. MMPs are a group of endopeptidases that degrade extracellular pro mechanisms in the genesis of inflammatory demyelination, such as recruitment, blood-brain barrier breakdown and myelin destruction are con be MMP-dependent processes. Aims of the study: 1. To investigate the levels of cytokine secreting blood and cerebrospinal fluid mononuclear cells (MNC) in patients with MS and controls; 2. To define further the cytokine disbalance in MS by examining the levels of cytokine receptors that are important in the net effect of cytokine function; 3. To study the levels of blood and CSF MNC in MS expressing mRNA of MMPs and their inhibitors; and 4. To examine the effects of IFN-[beta] on cytokines and MMPs in MS. Results: MS is associated with increased numbers of IL-6 and TNF-[alpha] secreting blood MNC and decreased numbers of IL-10 secreting cells compared to healthy subjects. In CSF, similar numbers of IL-6 and IL-10 secreting cells were observed in patients with MS and other neurological diseases (OND). Patients with MS also had higher levels of MMP-3, MMP-9 and tissue inhibitor of metalloproteinases-1 (TIMP-1) mRNA expressing blood MNC compared to controls. When patients with MS examined before vs. during 1 year of treatment with IFN-[beta], treatment resulted in decreased numbers of IL-6 and TNF-[alpha] secreting MNC

Immunological mechanisms play a central role in the pathogenesis sclerosis (MS). In this study two groups of related immune variables are cytokines and metalloproteinases (MMPs), which regulate each other's production and activation. Cytokines play key role in pathogenesis and treatments of Cytokines are regulatory proteins secreted by a variety of cells. The pleiotropic action of cytokines include numerous effects on cells of the immune modulating immune responses. Cytokine receptors are crucial for the net effect of cytokines. The balance between Th1 and Th2 type cytokines might determine whether the immune response in MS is detrimental or beneficial. MMPs are a group of endopeptidases that degrade extracellular pro mechanisms in the genesis of inflammatory demyelination, such as recruitment, blood-brain barrier breakdown and myelin destruction are con be MMP-dependent processes. Aims of the study: 1. To investigate the levels of cytokine secreting blood and cerebrospinal fluid mononuclear cells (MNC) in patients with MS and controls; 2. To define further the cytokine disbalance in MS by examining the levels of cytokine receptors that are important in the net effect of cytokine function; 3. To study the levels of blood and CSF MNC in MS expressing mRNA of MMPs and their inhibitors; and 4. To examine the effects of IFN-[beta] on cytokines and MMPs in MS. Results: MS is associated with increased numbers of IL-6 and TNF-[alpha] secreting blood MNC and decreased numbers of IL-10 secreting cells compared to healthy subjects. In CSF, similar numbers of IL-6 and IL-10 secreting cells were observed in patients with MS and other neurological diseases (OND). Patients with MS also had higher levels of MMP-3, MMP-9 and tissue inhibitor of metalloproteinases-1 (TIMP-1) mRNA expressing blood MNC compared to controls. When patients with MS examined before vs. during 1 year of treatment with IFN-[beta], treatment resulted in decreased numbers of IL-6 and TNF-[alpha] secreting MNC