ABSTRACT
Background: Pseudomonas aeruginosa is an important bacterial pathogen, particularly as a cause of nosocomial infections in hospitalized patients. Only few reports exist in which cystoscopes were implicated as an outbreak source. We describe an investigation into the cause of a sudden increase in the number of urinary tract infections (UTI) with P. aeruginosa in patients after cystoscopy. In addition, we share the lessons learned and measures taken to reduce the risk of similar infections in the future. Presentation of Case: Over a period of two weeks the urology outpatient department noticed a UTI in four patients following cystoscopy. An investigation was started for a common source of the outbreak in the urological treatment room. Additional screening of patients revealed a total of eleven males with P. aeruginosa UTI following cystoscopy. The infections were found to be due to a defective drying cabinet, which lacked an alarm signaling in case of loss of airflow. Amplified fragment length polymorphism (AFLP) analysis revealed that P. aeruginosa isolates from three patients and six isolates from environmental cultures (including cystoscopes from the drying cabinet) genotypically belonged to one strain. Discussion: The AFLP results suggest that contaminated cystoscopes caused P. aeruginosa UTI in 11 patients, with the drying cabinet as site of transfer of the infective strain. To our knowledge, this is the first report describing a malfunctioning drying cabinet as source of an outbreak following cystoscopy. Conclusion: In case of concomitant P. aeruginosa infections, cystoscopes and drying cabinets should be suspected as a potential source. Molecular techniques are helpful in investigating the epidemiology of an outbreak.
ABSTRACT
Currently, there is a nationwide outbreak of Mycoplasma pneumoniae infections. M. pneumoniae is a bacterium that can cause atypical pneumonia, especially in children and young adults, and does not respond to the standard antibiotics prescribed for pneumonia. In addition, the bacterium regularly causes extra-pulmonary symptoms. In our hospitals, we have admitted 100 patients (including 20 children) with M. pneumoniae since the fall of 2023, many of which were young and had severe clinical symptoms. It is important to recognize the clinical picture to start effective antibiotic treatment. In this clinical lesson, we will provide two examples of recently admitted patients and discuss the characteristics of all inpatients who have presented to our hospitals during this epidemic. Finally, we pay attention to antibiotic policy and antibiotic resistance.
Subject(s)
Anti-Bacterial Agents , Mycoplasma pneumoniae , Pneumonia, Mycoplasma , Humans , Netherlands/epidemiology , Anti-Bacterial Agents/therapeutic use , Mycoplasma pneumoniae/drug effects , Pneumonia, Mycoplasma/epidemiology , Pneumonia, Mycoplasma/drug therapy , Pneumonia, Mycoplasma/history , Child , Drug Resistance, Bacterial , Disease Outbreaks , Male , Female , AdultABSTRACT
The bioMérieux BIOFIRE Joint Infection (JI) Panel is a multiplex in vitro diagnostic test for the simultaneous and rapid (~1 h) detection of 39 potential pathogens and antimicrobial resistance (AMR) genes directly from synovial fluid (SF) samples. Thirty-one species or groups of microorganisms are included in the kit, as well as several AMR genes. This study, performed to evaluate the BIOFIRE JI Panel for regulatory clearance, provides data from a multicenter evaluation of 1,544 prospectively collected residual SF samples with performance compared to standard-of-care (SOC) culture for organisms or polymerase chain reaction (PCR) and sequencing for AMR genes. The BIOFIRE JI Panel demonstrated a sensitivity of 90.9% or greater for all but six organisms and a positive percent agreement (PPA) of 100% for all AMR genes. The BIOFIRE JI Panel demonstrated a specificity of 98.5% or greater for detection of all organisms and a negative percent agreement (NPA) of 95.7% or greater for all AMR genes. The BIOFIRE JI Panel provides an improvement over SOC culture, with a substantially shorter time to result for both organisms and AMR genes with excellent sensitivity/PPA and specificity/NPA, and is anticipated to provide timely and actionable diagnostic information for joint infections in a variety of clinical scenarios.
Subject(s)
Anti-Infective Agents , Arthritis, Infectious , Humans , Saccharomyces cerevisiae/genetics , Synovial Fluid/microbiology , Multiplex Polymerase Chain Reaction , Bacteria/genetics , Arthritis, Infectious/diagnosisABSTRACT
OBJECTIVE: Detection of the intracellular bacterium Coxiella burnetii, causative agent of chronic Q fever, is notoriously difficult. Diagnosis of and duration of antibiotic treatment for chronic Q fever is partly determined by detection of the bacterium with polymerase chain reaction (PCR). Fluorescence in situ hybridization (FISH) might be a promising technique for detecting C. burnetii in tissue samples from chronic Q fever patients, but its value in comparison with PCR is uncertain. We aim to assess the value of FISH for detecting C. burnetii in tissue of chronic Q fever patients. METHODS: FISH and PCR were performed on tissue samples from Dutch chronic Q fever patients collected during surgery or autopsy. Sensitivity, specificity, and overall diagnostic accuracy were calculated. Additionally, data on patient and disease characteristics were collected from electronic medical records. RESULTS: In total, 49 tissue samples from mainly vascular walls, heart valves, or placentas, obtained from 39 chronic Q fever patients, were examined by FISH and PCR. The sensitivity and specificity of FISH compared to PCR for detecting C. burnetii in tissue samples from chronic Q fever patients was 45.2% (95% confidence interval (CI), 27.3% - 64.0%) and 84.6% (95% CI, 54.6% - 98.1%), respectively. The overall diagnostic accuracy was 56.8% (95% CI, 42.2% - 72.3%). Two C. burnetii PCR negative placentas were FISH positive. Four FISH results (8.2%) were deemed inconclusive because of autofluorescence. CONCLUSION: With an overall diagnostic accuracy of 57.8%, we conclude that FISH has limited value in the routine diagnostics of chronic Q fever.
Subject(s)
Coxiella burnetii , Q Fever , Pregnancy , Female , Humans , Coxiella burnetii/genetics , Q Fever/diagnosis , Q Fever/microbiology , In Situ Hybridization, Fluorescence/methods , Heart Valves/microbiology , Anti-Bacterial AgentsABSTRACT
BACKGROUND: During the coronavirus disease 2019 (COVID-19) pandemic in the Netherlands it was noticed that very few blood cultures from COVID-19 patients turned positive with clinically relevant bacteria. This was particularly evident in comparison to the number of positive blood cultures during previous seasonal epidemics of influenza. This observation raised questions about the occurrence and causative microorganisms of bacteraemia in COVID-19 patients, especially in the perspective of the widely reported overuse of antibiotics and the rising rate of antibiotic resistance. METHODS: We conducted a retrospective cohort study on blood culture results in influenza A, influenza B and COVID-19 patients presenting to two hospitals in the Netherlands. Our main outcome consisted of the percentage of positive blood cultures. The percentage of clinically relevant blood cultures, isolated bacteria and 30-day all-cause mortality served as our secondary outcomes. RESULTS: A total of 1331 viral episodes were analysed in 1324 patients. There was no statistically significant difference (p = 0.47) in overall occurrence of blood culture positivity in COVID-19 patients (9.0, 95% CI 6.8-11.1) in comparison to influenza A (11.4, 95% CI 7.9-14.8) and influenza B patients (10.4, 95% CI 7.1-13.7,). After correcting for the high rate of contamination, the occurrence of clinically relevant bacteraemia in COVID-19 patients amounted to 1.0% (95% CI 0.3-1.8), which was statistically significantly lower (p = 0.04) compared to influenza A patients (4.0, 95% CI 1.9-6.1) and influenza B patients (3.0, 95% CI 1.2-4.9). The most frequently identified bacterial isolates in COVID-19 patients were Escherichia coli (n = 2) and Streptococcus pneumoniae (n = 2). The overall 30-day all-cause mortality for COVID-19 patients was 28.3% (95% CI 24.9-31.7), which was statistically significantly higher (p = <.001) when compared to patients with influenza A (7.1, 95% CI 4.3-9.9) and patients with influenza B (6.4, 95% CI 3.8-9.1). CONCLUSIONS: We report a very low occurrence of community-acquired bacteraemia amongst COVID-19 patients in comparison to influenza patients. These results reinforce current clinical guidelines on antibiotic management in COVID-19, which only advise utilization of antibiotics when a bacterial co-infection is suspected.
Subject(s)
Bacteremia/epidemiology , COVID-19/microbiology , Community-Acquired Infections/epidemiology , Influenza A virus , Influenza B virus , Influenza, Human/microbiology , SARS-CoV-2 , Adult , Aged , Aged, 80 and over , Anti-Bacterial Agents/therapeutic use , COVID-19/mortality , Female , Humans , Male , Middle Aged , Netherlands/epidemiology , Retrospective StudiesABSTRACT
Our study aim was to determine how a new clinical pathway, including PCR-based influenza point-of-care test (POCT), influences the hospitalisation costs of patients suspected of influenza presenting at the emergency department of a Dutch hospital during two consecutive influenza epidemics (2016-2017 and 2017-2018). Compared to mean costs per patient of 3661 in 2016-2017, the implementation of this new clinical pathway with influenza POCT in 2017 was associated with mean costs per influenza-positive patient of 2495 in 2017-2018 (P = .3). Our study suggests favourable economic results regarding a new clinical pathway with influenza POCT, reflecting a more efficient care of patients suspected of influenza presenting at the emergency department.
Subject(s)
Epidemics , Influenza, Human , Critical Pathways , Emergency Service, Hospital , Hospitals , Humans , Influenza, Human/diagnosis , Influenza, Human/epidemiology , Point-of-Care Systems , Point-of-Care TestingABSTRACT
INTRODUCTION: Coxiella burnetii, the causative pathogen of Q fever, is regularly detected in throat swabs from patients without serological evidence of Q fever infection. C. burnetii is also frequently found in bulk tank milk from dairy cows. We evaluated the false positivity rate of polymerase chain reaction (PCR) for C. burnetii DNA on throat swabs and investigated whether recent consumption of C. burnetii DNA-positive cow milk could contribute to this phenomenon. METHODS: C. burnetii PCR was performed on throat swabs obtained from patients in whom a throat swab was ordered for other diagnostic purposes; patients with community-acquired pneumonia (CAP); and healthy volunteers after consumption of commercial C. burnetii-containing cow milk products. RESULTS: C. burnetii DNA was found in 5.0% of throat swabs ordered for other diagnostic purposes and in 15.3% of throat swabs from CAP patients without serological evidence of Q fever pneumonia. The positive and negative predictive value of C. burnetii PCR on throat swabs for Q fever pneumonia were 66.7% (95% CI, 38.0-88.2) and 48.9% (95% CI, 41.3-54.6), respectively. After consumption of commercial C. burnetii-containing cow milk products, C. burnetii DNA could be detected in throat swabs for as long as 30â¯min after ingestion. CONCLUSION: C. burnetii PCR on throat swabs is of low diagnostic value for Q fever pneumonia and was false positive in 15.3% of CAP patients without Q fever pneumonia. Recent consumption of C. burnetii-containing products can influence the outcome of C. burnetii PCR on throat swabs. Therefore, diagnosis of C. burnetii infection should be made in combination with serology or PCR performed on blood.
Subject(s)
Coxiella burnetii/isolation & purification , DNA, Bacterial/analysis , Milk/microbiology , Pharynx/microbiology , Polymerase Chain Reaction/methods , Q Fever/diagnosis , Aged , Animals , False Positive Reactions , Female , Humans , Male , Middle AgedABSTRACT
PURPOSE: Coxiella burnetii has been suggested as a potential cause of B-cell non-Hodgkin lymphoma (B-NHL), as C. burnetii was detected in B-NHL tissues. To further investigate this potential relationship, we hypothesized that among subjects previously exposed to C. burnetii, the bacterium is more frequently detectable in tissues of patients with B-NHL (cases) compared to patients without B-NHL (controls). METHODS: We aimed to evaluate this hypothesis by assessing the presence of C. burnetii with polymerase chain reaction (PCR), immunofluorescence staining (IF) and fluorescent in-situ hybridization (FISH). Eligible patients were those previously exposed to C. burnetii. RESULTS: Samples were available for 13 cases and 16 controls. C. burnetii was demonstrated in tissues of 8/29 patients in total (28%), with either PCR, IF or FISH: in 5/13 cases (38%) and 3/16 controls (19%), p = 0.41. Negative and positive control samples were all negative and positive appropriately for all three diagnostic methods. CONCLUSIONS: In patients previously exposed to C. burnetii the bacterium was detected in tissue samples from subjects with and without B-NHL, without significant differences in the proportion positive samples. Therefore, we conclude that detection of C. burnetii in tissues of patients previously exposed to C. burnetii is a non-specific finding.
Subject(s)
Coxiella burnetii , Lymphoma, Non-Hodgkin/etiology , Q Fever/complications , Q Fever/microbiology , Adult , Aged , Aged, 80 and over , Comorbidity , Coxiella burnetii/genetics , Disease Susceptibility , Female , Fluorescent Antibody Technique , Humans , In Situ Hybridization, Fluorescence , Male , Middle Aged , Polymerase Chain ReactionABSTRACT
OBJECTIVES AND DESIGN: Non-Hodgkin lymphoma has been linked to infection with Coxiella burnetii, potentially through overproduction of IL-10 during infection with C. burnetii. MATERIALS AND METHODS: Description of a case report. RESULTS: We describe a patient with retroperitoneal non-Hodgkin lymphoma and vascular infection with C. burnetii. Immunofluorescence staining and fluorescence in situ hybridization targeting specific C. burnetii 16S rRNA were performed on the retroperitoneal lymphoma tissue sample obtained at diagnosis of NHL. Both were strongly positive for the presence of C. burnetii. CONCLUSIONS: This case provokes questions regarding a potential association between C. burnetii and NHL, and underlines the importance of further exploration of this association.
Subject(s)
Coxiella burnetii/isolation & purification , Lymphoma, Non-Hodgkin/diagnosis , Q Fever/diagnosis , Retroperitoneal Neoplasms/diagnosis , Coxiella burnetii/genetics , Fluorescent Antibody Technique , Humans , In Situ Hybridization, Fluorescence , Lymphoma, Non-Hodgkin/microbiology , Male , Middle Aged , Netherlands , Q Fever/microbiology , RNA, Bacterial/analysis , RNA, Ribosomal, 16S/analysis , Retroperitoneal Neoplasms/microbiologySubject(s)
Chlamydia Infections/transmission , Chlamydia/isolation & purification , Pneumonia, Bacterial/transmission , Zoonoses/transmission , Adult , Animals , Chlamydia/genetics , Community-Acquired Infections/transmission , DNA, Bacterial/isolation & purification , Female , Guinea Pigs , Humans , Male , Respiratory Insufficiency/etiologyABSTRACT
Trichomonas vaginalis is the most common sexually transmitted parasitical infection worldwide. T. vaginalis can carry a virus: Trichomonas vaginalis virus (TVV). To date, four TVV species have been described. Few studies have investigated TVV prevalence and its clinical importance. We have developed a nested reverse-transcriptase PCR, with novel, type specific primers to directly detect TVV RNA in T. vaginalis positive clinical samples. A total of 119T. vaginalis positive clinical samples were collected in Amsterdam and "s-Hertogenbosch, the Netherlands, from 2012 to 2016. For all samples T. vaginalis was genotyped using multi-locus sequence typing. The T. vaginalis positive samples segregated into a two-genotype population: type I (n=64) and type II (n=55). All were tested for TVV with the new TVV PCR. We detected 3 of the 4 TVV species. Sequencing of the amplified products showed high homology with published TVV genomes (82-100%). Half of the T. vaginalis clinical samples (n=60, 50.4%) were infected with one or more TVV species, with a preponderance for TVV infections in T. vaginalis type I (n=44, 73.3%). Clinical data was available for a subset of samples (n=34) and we observed an association between testing positive for (any) TVV and reporting urogenital symptoms (p=0.023). The nested RT-PCR allowed for direct detection of TVV in T. vaginalis positive clinical samples. This may be helpful in studies and clinical settings, since T. vaginalis disease and/or treatment outcome may be influenced by the protozoa"s virus.
Subject(s)
Polymerase Chain Reaction/methods , Totiviridae/isolation & purification , Trichomonas Vaginitis/parasitology , Trichomonas vaginalis/virology , Adult , DNA Primers , Female , Genome, Viral , Genotype , Humans , Male , Multilocus Sequence Typing , Netherlands/epidemiology , Phylogeny , RNA, Double-Stranded , Totiviridae/genetics , Trichomonas Vaginitis/epidemiologyABSTRACT
A disseminated peritoneal dialysis-related Mycobacterium abscessus infection is very rare. M. abscessus belongs to the rapidly growing mycobacteria and can be misidentified as a diphtheroid bacterium, which in our case delayed diagnosis and optimal treatment. Due to intrinsic resistance to most antimicrobials, therapeutic options in M. abscessus infections are limited. Infection often leads to catheter loss. A fatal outcome, like in our case, is not exceptional.
ABSTRACT
In the Netherlands, the number of cases of infection with New Delhi metallo-beta-lactamase (NDM)-positive Enterobacteriaceae is low. Here, we report an outbreak of NDM-1-producing Klebsiella pneumoniae infection in a Dutch hospital with interspecies transfer of the resistance plasmid and unexpected occurrence in other unrelated health care centers (HCCs). Next-generation sequencing was performed on 250 carbapenemase-producing Enterobacteriaceae isolates, including 42 NDM-positive isolates obtained from 29 persons at the outbreak site. Most outbreak isolates were K. pneumoniae (n = 26) and Escherichia coli (n = 11), but 5 isolates comprising three other Enterobacteriaceae species were also cultured. The 26 K. pneumoniae isolates had sequence type 873 (ST873), as did 7 unrelated K. pneumoniae isolates originating from five geographically dispersed HCCs. The 33 ST873 isolates that clustered closely together using whole-genome multilocus sequence typing (wgMLST) carried the same plasmids and had limited differences in the resistome. The 11 E. coli outbreak isolates showed great variety in STs, did not cluster using wgMLST, and showed considerable diversity in resistome and plasmid profiles. The blaNDM-1 gene-carrying plasmid present in the ST873 K. pneumoniae isolates was found in all the other Enterobacteriaceae species cultured at the outbreak location and in a single E. coli isolate from another HCC. We describe a hospital outbreak with an NDM-1-producing K. pneumoniae strain from an unknown source that was also found in patients from five other Dutch HCCs in the same time frame without an epidemiological link. Interspecies transfer of the resistance plasmid was observed in other Enterobacteriaceae species isolated at the outbreak location and in another HCC.
Subject(s)
Cross Infection/epidemiology , Disease Outbreaks , Enterobacteriaceae/enzymology , Gene Transfer, Horizontal , Klebsiella Infections/epidemiology , Plasmids/analysis , beta-Lactamases/genetics , Cross Infection/microbiology , Enterobacteriaceae/classification , Enterobacteriaceae/genetics , Enterobacteriaceae/isolation & purification , Genotype , Health Facilities , Humans , Klebsiella Infections/microbiology , Multilocus Sequence Typing , Netherlands/epidemiologyABSTRACT
BACKGROUND: Breast cancer is a heterogeneous disease with various histological features and molecular markers. These are utilized for the prediction of clinical outcome and therapeutic decision making. In addition to well established markers such as HER2 overexpression and estrogen and progesterone receptor (ER and PR) status, chromosomal instability is evolving as an important hallmark of cancers. The HER2/TOP2A locus is of great importance in breast cancer. The copy number variability at this locus has been proposed to be a marker for the degree of chromosomal instability. We therefore developed a Single Nucleotide Polymorphism (SNP) assay to evaluate allelic imbalance at the HER2/TOP2A locus in three different entities of primary breast tumors. METHODS: Eleven SNPs were carefully selected and detected by real time PCR using DNA extracted from paired (histologically normal and tumor) paraffin-embedded tissues. Primary breast tumors of 44 patients were included, 15 tumors with HER2 overexpression, 16 triple negative tumors, defined by the absence of HER2 overexpression and a negative ER and PR status and 13 ER and PR positive tumors without HER2 overexpression. As controls, histologically normal breast tissues from 10 patients with no breast tumor were included. RESULTS: Allelic imbalance was observed in 13/15 (87 %) HER2 positive tumors, the remaining 2 being inconclusive. Of the 16 triple negative tumors, 12 (75 %) displayed instability, 3 (19 %) displayed no instability, and 1 was inconclusive. Of the 13 hormone receptor positive tumors, 5 (38 %) displayed allelic imbalance, while 8 did not. CONCLUSIONS: We conclude that the SNP assay is suitable for rapid testing of allelic (im)balance at the HER2/TOP2A locus using paraffin-embedded tissues. Based on allelic imbalance at this locus, both triple negative and ER and PR positive breast tumors can be subcategorized. The clinical relevance of the allelic (im)balance status at the HER2/TOP2A locus in breast cancer is subject of future study. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/2086062232155220.
Subject(s)
Allelic Imbalance , Antigens, Neoplasm/genetics , Biomarkers, Tumor/genetics , Breast Neoplasms/genetics , DNA Topoisomerases, Type II/genetics , DNA-Binding Proteins/genetics , Polymorphism, Single Nucleotide , Receptor, ErbB-2/genetics , Triple Negative Breast Neoplasms/genetics , Adult , Aged , Biomarkers, Tumor/analysis , Biopsy , Breast Neoplasms/chemistry , Breast Neoplasms/pathology , Female , Genetic Predisposition to Disease , Humans , Immunohistochemistry , In Situ Hybridization , Middle Aged , Paraffin Embedding , Phenotype , Poly-ADP-Ribose Binding Proteins , Real-Time Polymerase Chain Reaction , Receptor, ErbB-2/analysis , Receptors, Estrogen/analysis , Receptors, Progesterone/analysis , Retrospective Studies , Triple Negative Breast Neoplasms/chemistry , Triple Negative Breast Neoplasms/pathology , Young AdultABSTRACT
A large community outbreak of Q fever occurred in the Netherlands in the period 2007 to 2010. Some of the infected patients developed chronic Q fever, which typically includes pathogen dissemination to predisposed cardiovascular sites, with potentially fatal consequences. To identify the immune mechanisms responsible for ineffective clearance of Coxiella burnetii in patients who developed chronic Q fever, we compared serum concentrations of 47 inflammation-associated markers among patients with acute Q fever, vascular chronic Q fever, and past resolved Q fever. Serum levels of gamma interferon were strongly increased in acute but not in vascular chronic Q fever patients, compared to past resolved Q fever patients. Interleukin-18 levels showed a comparable increase in acute as well as vascular chronic Q fever patients. Additionally, vascular chronic Q fever patients had lower serum levels of gamma interferon-inducible protein 10 (IP-10) and transforming growth factor ß (TGF-ß) than did acute Q fever patients. Serum responses for these and other markers indicate that type I immune responses to C. burnetii are affected in chronic Q fever patients. This may be attributed to an affected immune system in cardiovascular patients, which enables local C. burnetii replication at affected cardiovascular sites.
Subject(s)
Interferon-gamma/blood , Q Fever/immunology , Q Fever/pathology , Serum/chemistry , Adult , Aged , Aged, 80 and over , Chemokine CXCL10/blood , Female , Humans , Male , Middle Aged , Netherlands/epidemiology , Q Fever/epidemiology , Retrospective Studies , Transforming Growth Factor beta/blood , Young AdultABSTRACT
The aim of this study was to describe specific histological findings of the Coxiella burnetii-infected aneurysmal abdominal aortic wall. Tissue samples of the aneurysmal abdominal aortic wall from seven patients with chronic Q fever and 15 patients without evidence of Q fever infection were analysed and compared. Chronic Q fever was diagnosed using serology and tissue PCR analysis. Histological sections were stained using haematoxylin and eosin staining, Elastica van Gieson staining and immunohistochemical staining for macrophages (CD68), T lymphocytes (CD3), T lymphocyte subsets (CD4 and CD8) and B lymphocytes (CD20). Samples were scored by one pathologist, blinded for Q fever status, using a standard score form. Seven tissue samples from patients with chronic Q fever and 15 tissue samples from patients without Q fever were collected. Four of seven chronic Q fever samples showed a necrotizing granulomatous response of the vascular wall, which was characterized by necrotic core of the arteriosclerotic plaque (P = 0.005) and a presence of high numbers of macrophages in the adventitia (P = 0.007) distributed in typical palisading formation (P = 0.005) and surrounded by the presence of high numbers of T lymphocytes located diffusely in media and adventitia. Necrotizing granulomas are a histological finding in the C. burnetii-infected aneurysmal abdominal aortic wall. Chronic Q fever should be included in the list of infectious diseases with necrotizing granulomatous response, such as tuberculosis, cat scratch disease and syphilis.
Subject(s)
Aorta, Abdominal/microbiology , Aorta, Abdominal/pathology , Aortic Aneurysm, Abdominal/microbiology , Aortic Aneurysm, Abdominal/pathology , Q Fever/microbiology , Q Fever/pathology , Aged , Aged, 80 and over , B-Lymphocytes/pathology , Coxiella burnetii/isolation & purification , Female , Granuloma/pathology , Humans , Macrophages/pathology , Male , Middle Aged , Necrosis , Prospective Studies , Retrospective Studies , T-Lymphocytes/pathologyABSTRACT
OBJECTIVES: The aim of this study was to estimate the seroprevalence of Q fever and prevalence of chronic Q fever in patients with abdominal aortic and/or iliac disease after the Q fever outbreak of 2007-2010 in the Netherlands. METHODS: In November 2009, an ongoing screening program for Q fever was initiated. Patients with abdominal aortic and/or iliac disease were screened for presence of IgM and IgG antibodies to phase I and II antigens of Coxiella burnetii using immunofluorescence assay and presence of C. burnetii DNA in sera and/or vascular wall tissue using polymerase chain reaction (PCR). RESULTS: A total of 770 patients with abdominal aortic and/or iliac disease were screened. Antibodies against C. burnetii were detected in 130 patients (16.9%), of which 40 (30.8%) patients showed a serological profile of chronic Q fever. Three patients presented with acute Q fever, one of which developed to chronic Q fever over time. The number of aneurysm-related acute complications in patients with chronic Q fever was significantly higher compared to patients negative for Q fever (p = 0.013); 9.0% (30/333) vs. 30.0% (6/20). Eight out of 46 patients with past resolved Q fever (8/46, 17.4%) presented with aneurysm-related acute complications (no significant difference). CONCLUSION: The prevalence of chronic Q fever in C. burnetii seropositive patients with abdominal aortic and/or iliac disease living in an epidemic area in the Netherlands is remarkably high (30.8%). Patients with an aneurysm and chronic Q fever present more often with an aneurysm-related acute complication compared to patients without evidence of Q fever infection.
Subject(s)
Aortic Aneurysm/epidemiology , Coxiella burnetii/isolation & purification , Iliac Aneurysm/epidemiology , Q Fever/epidemiology , Aged , Antibodies, Bacterial/blood , Aortic Aneurysm/diagnosis , Aortic Aneurysm/microbiology , Comorbidity , DNA, Bacterial/blood , DNA, Bacterial/isolation & purification , Disease Outbreaks , Female , Humans , Iliac Aneurysm/complications , Iliac Aneurysm/diagnosis , Iliac Aneurysm/microbiology , Immunoglobulin G/blood , Immunoglobulin M/blood , Logistic Models , Male , Netherlands/epidemiology , Prevalence , Q Fever/blood , Q Fever/diagnosis , Risk Factors , Seroepidemiologic StudiesABSTRACT
BACKGROUND: After the largest outbreaks of Q fever ever recorded in history occurred in the Netherlands, concern arose that Coxiella may be transmitted via donated tissues of latent or chronically infected donors. The Dutch Health Council recently advised to screen tissue donors, donating high risk tissues, for Coxiella infection. METHODS: After validation of an enzyme immunoassay (EIA) test for IgG antibodies against phase 2 of C. burnetii for use on post-mortem samples, serum samples of 1033 consecutive Dutch post-mortem tissue donors were tested for IgG antibodies against phase 2 of C. burnetii. Confirmation of reactive results was done by immunofluorescence assay (IFA). All available tissues (corneas, heart valves, skin and bone marrow) from donors with IgG reactivity were tested for presence of Coxiella DNA by PCR. Risk factors for IgG reactivity were investigated. RESULTS: After validation of the tests for use on post-mortem samples, 50/1033 donors (4.8%) screened positive for phase 2 anti-Coxiella IgG by EIA, and 31 were confirmed by IFA (3.0%). One donor showed a serological profile compatible with chronic infection. All tested tissues (25 corneas, 6 heart valves, 4 skin and 3 bone marrow) from donors with IgG reactivity tested negative for the presence of Coxiella DNA. Except for living in a postal code area with a high number of Q fever notifications, no risk factors for IgG reactivity were found. CONCLUSIONS: The strong correlation between notifications and seroprevalence confirms that the used assays are sufficiently specific for use on post-mortem samples, although one has to be aware of differences between batches. Thus, this study provides a validated method for screening tissue donors for infection with Coxiella burnetii that can be used in future outbreaks.
Subject(s)
Coxiella burnetii/isolation & purification , DNA, Bacterial/analysis , Tissue Donors , Adult , Aged , Autopsy , Chronic Disease , Communicable Diseases/epidemiology , Coxiella burnetii/immunology , Disease Outbreaks , Female , Humans , Immunoenzyme Techniques , Immunoglobulin G/analysis , Male , Middle Aged , Netherlands/epidemiology , Polymerase Chain Reaction/methods , Q Fever/epidemiology , Risk , Risk Factors , Seroepidemiologic StudiesABSTRACT
BACKGROUND: Resistance to carbapenem antibiotics is emerging worldwide among Enterobacteriaceae. To prevent hospital transmission due to unnoticed carriage of carbapenemase producing micro-organisms in newly admitted patients, or follow-up of patients in an outbreak setting, a molecular screening method was developed for detection of the most prevalent carbapenemase genes; blaOXA-48, blaVIM, blaIMP, blaNDM and blaKPC. METHODS: A real-time multiplex PCR assay was evaluated using a collection of 86 Gram negative isolates, including 62 carbapenemase producers. Seven different laboratories carried out this method and used the assay for detection of the carbapenemase genes on a selection of 20 isolates. RESULTS: Both sensitivity and specificity of the multiplex PCR assay was 100%, as established by results on the strain collection and the inter-laboratory comparisons. CONCLUSIONS: In this study, we present a multiplex real-time PCR that is a robust, reliable and rapid method for the detection of the most prevalent carbapenemases blaOXA-48, blaVIM, blaIMP, blaNDM and blaKPC, and is suitable for screening of broth cultured rectal swabs and for identification of carbapenemase genes in cultures.
Subject(s)
Bacterial Proteins/genetics , Carbapenems , Drug Resistance, Bacterial/genetics , Enterobacteriaceae/genetics , Multiplex Polymerase Chain Reaction , beta-Lactamases/genetics , Humans , Real-Time Polymerase Chain Reaction , Sensitivity and SpecificityABSTRACT
OBJECTIVES: The objective of this study was to investigate the antibiotic resistance patterns, including methicillin resistance, inducible macrolide-lincosamide-streptogramin B (MLSB ) resistance and Panton-Valentine leukocidin (PVL) toxin gene carriage among hospital-acquired Staphylococcus aureus (HA-SA) and community-acquired S. aureus (CA-SA), in Beira, Mozambique. METHODS: In 2010-2011, two prospective surveillance studies were conducted on post-operative and burn wound infections at the Central Hospital of Beira and on skin and soft tissue abscesses at the São Lucas Health Centre. We cultured pus samples, identified suspected S. aureus isolates and performed antimicrobial susceptibility testing, including detection of MLSB resistance. Real-time polymerase chain reaction was used to detect mecA, Martineau and PVL genes. RESULTS: The prevalence of hospital-acquired methicillin-resistant S. aureus (HA-MRSA) infection among 53 inpatients was 15.1%; the prevalence of community-acquired methicillin-resistant S. aureus (CA-MRSA) infection among 100 outpatients was 1.0%. Inducible MLSB resistance was present in 41.7% and 10.7% of HA-SA and CA-SA isolates, respectively. PVL toxin gene was detected in 81.1% of methicillin-susceptible S. aureus (MSSA) compared with 11.1% of methicillin-resistant S. aureus. CONCLUSIONS: Our study shows, for the first time in Mozambique, the emergence of HA-MRSA. The prevalence of CA-MRSA was low, whereas the rate of PVL toxin gene carriage in MSSA was high. The high rate of inducible MLSB resistance indicates the importance of performing routine D-tests. Overall, our results show the need of strengthening laboratory facilities to provide microbiological data for both directed therapy and surveillance.