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1.
Rev Chilena Infectol ; 38(3): 384-392, 2021 Jun.
Artigo em Espanhol | MEDLINE | ID: mdl-34479296

RESUMO

This narrative review includes published studies of stillbirth classification methods and their efficiency in identifying ascending bacterial infection (ABI), as a cause of fetal death (FD), by searching PubMed, Cochrane, Embase, ScienceDirect, Wiley Online Library, Scielo. Many children die before birth around the world and it has not been possible to reduce FD because the methods used have not been adequate and because ABI, the most frequent cause of FD in a public hospital in Chile, is not diagnosed. Systems using clinical, laboratory and placental study data, INCODE, CORM, are more efficient in identifying ABI as the origin of FD. Specific markers of infection/placental inflammation, histologic chorioamnionitis/acute funitis have been shown to be more efficient in diagnosing ABI than fetal autopsy, that amniotic fluid culture is more efficient than placental culture for detect microbial invasion of the amniotic cavity and that the cord blood sample is efficient for the etiological diagnosis of the infection. The knowledge of the ABI as the initial cause of FD helps to develop guidelines and norms for preventing FD due to this condition.


Assuntos
Infecções Bacterianas , Placenta , Líquido Amniótico , Infecções Bacterianas/diagnóstico , Criança , Feminino , Morte Fetal , Humanos , Laboratórios , Gravidez
2.
Crit Care ; 25(1): 281, 2021 08 05.
Artigo em Inglês | MEDLINE | ID: mdl-34353339

RESUMO

BACKGROUND: Procalcitonin (PCT) and C-reactive protein (CRP) were previously shown to have value for the detection of secondary infections in critically ill COVID-19 patients. However, since the introduction of immunomodulatory therapy, the value of these biomarkers is unclear. We investigated PCT and CRP kinetics in critically ill COVID-19 patients treated with dexamethasone with or without tocilizumab, and assessed the value of these biomarkers to detect secondary bacterial infections. METHODS: In this prospective study, 190 critically ill COVID-19 patients were divided into three treatment groups: no dexamethasone, no tocilizumab (D-T-), dexamethasone, no tocilizumab (D+T-), and dexamethasone and tocilizumab (D+T+). Serial data of PCT and CRP were aligned on the last day of dexamethasone treatment, and kinetics of these biomarkers were analyzed between 6 days prior to cessation of dexamethasone and 10 days afterwards. Furthermore, the D+T- and D+T+ groups were subdivided into secondary infection and no-secondary infection groups to analyze differences in PCT and CRP kinetics and calculate detection accuracy of these biomarkers for the occurrence of a secondary infection. RESULTS: Following cessation of dexamethasone, there was a rebound in PCT and CRP levels, most pronounced in the D+T- group. Upon occurrence of a secondary infection, no significant increase in PCT and CRP levels was observed in the D+T- group (p = 0.052 and p = 0.08, respectively). Although PCT levels increased significantly in patients of the D+T+ group who developed a secondary infection (p = 0.0003), this rise was only apparent from day 2 post-infection onwards. CRP levels remained suppressed in the D+T+ group. Receiver operating curve analysis of PCT and CRP levels yielded area under the curves of 0.52 and 0.55, respectively, which are both markedly lower than those found in the group of COVID-19 patients not treated with immunomodulatory drugs (0.80 and 0.76, respectively, with p values for differences between groups of 0.001 and 0.02, respectively). CONCLUSIONS: Cessation of dexamethasone in critically ill COVID-19 patients results in a rebound increase in PCT and CRP levels unrelated to the occurrence of secondary bacterial infections. Furthermore, immunomodulatory treatment with dexamethasone and tocilizumab considerably reduces the value of PCT and CRP for detection of secondary infections in COVID-19 patients.


Assuntos
Anticorpos Monoclonais Humanizados/uso terapêutico , Infecções Bacterianas/diagnóstico , COVID-19/tratamento farmacológico , Coinfecção/diagnóstico , Dexametasona/uso terapêutico , Idoso , Proteína C-Reativa/análise , COVID-19/complicações , Estado Terminal , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Países Baixos , Pró-Calcitonina/análise , Estudos Prospectivos
3.
Pan Afr Med J ; 38: 383, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34381527

RESUMO

Introduction: lower respiratory tract infections (LRTIs) are infections involving the trachea, primary bronchi and lungs. People with LRTIs typically experience coughs as the primary symptoms; however, shortness of breath, weakness, fever and fatigue may be coupled with the cough. It is common among the aged, children under five and the immune-suppressed. Persons with symptoms suggestive of pulmonary tuberculosis (TB) may have tuberculosis, other respiratory tract infection or co-infection of tuberculosis and other respiratory pathogens. This study aimed to identify the presence of pathogens in sputum of suspected tuberculosis cases and their antimicrobial resistance patterns. Methods: this was a retrospective study conducted from September 2018 to November 2019 at Tamale Public Health Laboratory. Sputum or gastric lavage samples were collected from persons with suspected clinical presentations of TB and/or LRTI. These samples were cultured using standard microbiological protocols and antimicrobial susceptibility test performed on the positive cultures by Kirby-Bauer disc diffusion method. Molecular identification of M. tuberculosis was performed on all the suspected TB cases using GeneXpert mycobacterium tuberculosis/rifampin (MTB/RIF) assay. Results: during the study period, there were 264 cases of which 49.2% were males and 50.8% were females. Positive cases for culture were 47.3%. Out of the 264 cases, 186 (70.5%) were suspected TB with 51.6% being positive for culture, 6.5% positive for M. tuberculosis (GeneXpert confirmed) and 3.8% co-infection of TB with other bacteria pathogens. Klebsiella spp. (35/125; 28%) and Pseudomonas spp. (19/125; 15.2%) were the most predominant pathogens isolated. There was no significant difference in detection of bacteria in males and females (p=0.89), however individuals with suspected TB were significantly infected with other bacterial species than the unsuspected individuals (p=0.03). Almost all the isolates showed high susceptibility towards carbapenem (meropenem) and high resistance towards the third generation cephalosporins (cefotaxime and ceftriaxone). Conclusion: this study highlights the need to test individuals with classical symptoms of LRTIs for other bacterial infections other than TB only. Sputum culture is recommended for all suspected tuberculosis cases to provide accurate laboratory diagnosis to LRTIs and mitigate unnecessary use of antimicrobials.


Assuntos
Bactérias/isolamento & purificação , Infecções Bacterianas/microbiologia , Infecções Respiratórias/microbiologia , Escarro/microbiologia , Adulto , Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/epidemiologia , Estudos Transversais , Feminino , Lavagem Gástrica , Humanos , Masculino , Testes de Sensibilidade Microbiana , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/epidemiologia , Estudos Retrospectivos , Adulto Jovem
4.
Appl Microbiol Biotechnol ; 105(16-17): 6245-6255, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34415392

RESUMO

Diagnosis of bacterial infections until today mostly relies on conventional microbiological methods. The resulting long turnaround times can lead to delayed initiation of adequate antibiotic therapy and prolonged periods of empiric antibiotic therapy (e.g., in intensive care medicine). Therewith, they contribute to the mortality of bacterial infections and the induction of multidrug resistances. The detection of species specific volatile organic compounds (VOCs) emitted by bacteria has been proposed as a possible diagnostic approach with the potential to serve as an innovative point-of-care diagnostic tool with very short turnaround times. A range of spectrometric methods are available which allow the detection and quantification of bacterial VOCs down to a range of part per trillion. This narrative review introduces the application of spectrometric analytical methods for the purpose of detecting VOCs of bacterial origin and their clinical use for diagnosing different infectious conditions over the last decade. KEY POINTS: • Detection of VOCs enables bacterial differentiation in various medical conditions. • Spectrometric methods may function as point-of-care diagnostics in near future.


Assuntos
Infecções Bacterianas , Compostos Orgânicos Voláteis , Bactérias , Infecções Bacterianas/diagnóstico , Humanos , Técnicas Microbiológicas , Análise Espectral
5.
Talanta ; 233: 122610, 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34215094

RESUMO

Recently, antibiotic resistant has become a serious public health concern, which warrants new generations of antibiotics to be developed. Pharmacodynamic evaluation is crucial in drug discovery processes. Despite numerous advanced imaging systems are available nowadays, technologies for the sensitive in vivo diagnosis of bacterial infections and direct visualization of drug efficacy are yet to be developed. In this study, we have developed novel near-infrared (NIR) fluorogenic probes. These probes are dark in solution but highly fluorescent when bound to the cognate reporter, fluorogen-activating protein (FAP). We established the in vivo bacterial infection model using FAP_dH6.2 recombinantly expressed E. coli and applied this NIR fluoromodule-based system for diagnosing bacterial infections and monitoring disease progressions and its responses to a type of antibiotics through classic mechanism of membrane lysis. This NIR fluoromodule-based system will discover new information on bacterial infections and identify newer antibacterial entities.


Assuntos
Infecções Bacterianas , Corantes Fluorescentes , Antibacterianos/farmacologia , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/tratamento farmacológico , Escherichia coli/genética , Humanos , Proteínas
6.
Medicine (Baltimore) ; 100(27): e26596, 2021 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-34232210

RESUMO

ABSTRACT: The objectives of this study were to understand the clinical presentations of febrile young infants with severe bacterial infection (SBI), and to investigate the pathogen variations throughout the vaccine era and after antenatal group B Streptococcus (GBS) screening.All infants < 90 days old with a body temperature of ≥38.0°C and admitted to the emergency department were retrospectively enrolled in our study. SBI was defined as a positive culture of urine, blood, or cerebrospinal fluid. All clinical variables were analyzed and compared between the SBI group and the non-SBI group, to identify the relevant risk factors for SBI in infants with pyrexia.A total of 498 infants were studied, 279 of whom (56%) had SBI. The body temperature at triage was higher in the SBI group, and the difference was highly obvious in the neonatal group. White blood cell count and C-reactive protein levels were both significantly higher in the SBI group (P < .05), whereas neutrophil percentage and band percentage demonstrated no significant differences. Escherichia coli was the most common pathogen and plasmid-mediated extended-spectrum lactamases were detected in up to 9.1%. GBS was detected in 16 cases of bacteremia (6 cases with concurrent meningitis).The body temperature at triage may provide a clue for differentiating sick babies, especially in the neonatal group. Complete serum analysis is required for infection survey, especially white blood cell and C-reactive protein. Escherichia coli is the most common pathogen, and clinician should raise awareness of drug resistance in some patients. The prevalence of GBS infection in the young infant group remains high after routine antenatal GBS screening.


Assuntos
Bactérias/isolamento & purificação , Infecções Bacterianas/epidemiologia , Serviço Hospitalar de Emergência/estatística & dados numéricos , Febre/etiologia , Infecções Bacterianas/complicações , Infecções Bacterianas/diagnóstico , Biomarcadores/sangue , Proteína C-Reativa/metabolismo , Feminino , Febre/sangue , Febre/epidemiologia , Seguimentos , Humanos , Incidência , Lactente , Recém-Nascido , Masculino , Estudos Retrospectivos , Taiwan/epidemiologia
8.
Emerg Med J ; 38(9): 685-691, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34289966

RESUMO

BACKGROUND: Guidelines recommend maximal efforts to obtain blood and sputum cultures in patients with COVID-19, as bacterial coinfection is associated with worse outcomes. The aim of this study was to evaluate the yield of bacteriological tests, including blood and sputum cultures, and the association of multiple biomarkers and the Pneumonia Severity Index (PSI) with clinical and microbiological outcomes in patients with COVID-19 presenting to the emergency department (ED). METHODS: This is a substudy of a large observational cohort study (PredictED study). The PredictED included adult patients from whom a blood culture was drawn at the ED of Haga Teaching Hospital, The Netherlands. For this substudy, all patients who tested positive for SARS-CoV-2 by PCR in March and April 2020 were included. The primary outcome was the incidence of bacterial coinfection. We used logistic regression analysis for associations of procalcitonin, C reactive protein (CRP), ferritin, lymphocyte count and PSI score with a severe disease course, defined as intensive care unit admission and/or 30-day mortality. The area under the receiver operating characteristics curve (AUC) quantified the discriminatory performance. RESULTS: We included 142 SARS-CoV-2 positive patients. On presentation, the median duration of symptoms was 8 days. 41 (29%) patients had a severe disease course and 24 (17%) died within 30 days. The incidence of bacterial coinfection was 2/142 (1.4%). None of the blood cultures showed pathogen growth while 6.3% was contaminated. The AUCs for predicting severe disease were 0.76 (95% CI 0.68 to 0.84), 0.70 (0.61 to 0.79), 0.62 (0.51 to 0.74), 0.62 (0.51 to 0.72) and 0.72 (0.63 to 0.81) for procalcitonin, CRP, ferritin, lymphocyte count and PSI score, respectively. CONCLUSION: Blood cultures appear to have limited value while procalcitonin and the PSI appear to be promising tools in helping physicians identify patients at risk for severe disease course in COVID-19 at presentation to the ED.


Assuntos
Infecções Bacterianas/diagnóstico , Técnicas Bacteriológicas/métodos , COVID-19/diagnóstico , Coinfecção/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Infecções Bacterianas/sangue , Infecções Bacterianas/complicações , Infecções Bacterianas/microbiologia , Técnicas Bacteriológicas/estatística & dados numéricos , Proteína C-Reativa/análise , COVID-19/sangue , COVID-19/complicações , COVID-19/virologia , Teste de Ácido Nucleico para COVID-19 , Coinfecção/sangue , Coinfecção/epidemiologia , Coinfecção/microbiologia , Serviço Hospitalar de Emergência , Feminino , Ferritinas/sangue , Humanos , Incidência , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Países Baixos/epidemiologia , Pró-Calcitonina/sangue , Prognóstico , Curva ROC , Estudos Retrospectivos , SARS-CoV-2/genética , SARS-CoV-2/isolamento & purificação , Índice de Gravidade de Doença
10.
Lancet Digit Health ; 3(8): e507-e516, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34325854

RESUMO

BACKGROUND: Acute febrile illness is one of the main reasons for outpatient hospital visits worldwide. However, differential diagnosis between bacterial and viral causes is challenging and misdiagnosis can result in antimicrobial overuse and hinder prompt treatment. We aimed to build and validate a diagnostic model to discriminate bacterial from viral infection in acute febrile illness by evaluating the expression of potential classifier host genes. METHODS: In this multicentre discovery and validation study, we included patients aged 14-85 years with acute febrile illness (fever for ≤14 days, axillary temperature of ≥38°C, and confirmed bacterial infection, viral infection, or non-infectious inflammatory disease), and healthy control participants (no significant medical history and no fever within the past 90 days) from four hospitals in Shandong province, China. Patients from the first hospital were divided into the screening, discovery, and internal validation groups, and patients from the three other hospitals comprised the external validation group. We measured expression of candidate genes in peripheral blood by RT-PCR, and patients for whom a successful RT-PCT result was recorded were included in the next-step analysis. For patients from the first hospital, those enrolled during the early phase of the study were assigned to the screening group, which was used to identify the optimal transcripts (IFI44L and PI3) for discrimination between bacterial and viral infections by screening four candidate genes (FAM89A, IFI44L, PI3, and ITGB2) by RT-PCR. The remaining patients were then randomly assigned (1:1) to discovery and internal validation groups by time of admission and blood drawing via the equidistant random sampling method. A logistic regression model integrating the mRNA levels of IFI44L and PI3 was built by use of the discovery group, and the diagnostic performance of the model was evaluated in the internal and external validation groups using area under the receiver operating curve (AUC), sensitivity, and specificity. FINDINGS: Between March 1, 2018, and Aug 31, 2019, we assessed 1658 individuals for inclusion in the study. After exclusion of ineligible participants, 458 participants were enrolled (178 patients with acute febrile illness caused by bacterial infection, 212 with acute febrile illness caused by viral infection, 38 with non-infectious inflammatory diseases, and 30 healthy controls). The 390 patients with bacterial or viral infections were assigned to one of four groups: screening (n=64, 33 with bacterial infections and 31 with viral infections), discovery (n=124, 55 with bacterial infections and 69 with viral infections), internal validation (n=124, 55 with bacterial infections and 69 with viral infections), and external validation (n=78, 35 with bacterial infections and 43 with viral infections). Of the four candidate host genes (FAM89A, IFI44L, PI3, and ITGB2), IFI44L and PI3 showed the most discriminative expression pattern and were used to build the logistic regression model. We established the optimal cutoff of the bacterial infection likelihood score to be 0·547598. With the diagnostic result from the gold standard tests (culture and PCR) as the reference, the two-transcript classifier model had an AUC of 0·969 (95% CI 0·937-1·000), sensitivity of 0·891 (0·782-0·949), and specificity of 0·971 (0·900-0·992) to discriminate bacterial and viral infections in the internal validation group. The model showed similar results in the external validation group (AUC 0·986, 95% CI 0·968-1·000; sensitivity 0·857, 0·706-0·937; and specificity 0·954, 0·845-0·987). INTERPRETATION: IFI44L and PI3 transcripts, measured by RT-PCR, are robust classifiers to discriminate bacterial from viral infection in acute febrile illness. This two-transcript biomarker has the potential to be transformed into a commercial panel and applied universally. FUNDING: None.


Assuntos
Bactérias , Infecções Bacterianas/diagnóstico , Febre/diagnóstico , Programas de Rastreamento/métodos , Modelos Biológicos , Viroses/diagnóstico , Vírus , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Área Sob a Curva , Bactérias/crescimento & desenvolvimento , Infecções Bacterianas/metabolismo , Infecções Bacterianas/microbiologia , Biomarcadores/metabolismo , China , Diagnóstico Diferencial , Feminino , Febre/metabolismo , Febre/microbiologia , Febre/virologia , Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Curva ROC , Reprodutibilidade dos Testes , Viroses/metabolismo , Viroses/virologia , Vírus/crescimento & desenvolvimento , Adulto Jovem
11.
BMC Infect Dis ; 21(1): 509, 2021 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-34059003

RESUMO

BACKGROUND: Readily-available diagnostics do not reliably discriminate between viral and bacterial pediatric uncomplicated pneumonia, both of which are common. Some have suggested that assessment of pneumococcal carriage could be used to identify those children with bacterial pneumonia. The objective of this study was to determine if nasopharyngeal pneumococcal colonization patterns differed between children with definite viral disease, definite bacterial disease, and respiratory disease of indeterminate etiology. METHODS: Three groups of subjects were recruited: children with critical respiratory illness, previously healthy children with respiratory illness admitted to the ward, and previously healthy children diagnosed in the emergency department with non-severe pneumonia. Subjects were categorized as follows: a) viral infection syndrome (eg. bronchiolitis), b) bacterial infection syndrome (ie. pneumonia complicated by effusion/empyema), or c) 'indeterminate' pneumonia. Subjects' nasopharyngeal swabs underwent quantitative PCR testing for S. pneumoniae. Associations between categorical variables were determined with Fisher's exact, chi-square, or logistic regression, as appropriate. Associations between quantitative genomic load and categorical variables was determined by linear regression. RESULTS: There were 206 children in Group 1, 122 children in Group 2, and 179 children in Group 3. Only a minority (227/507, 45%) had detectable pneumococcal carriage; in those subjects, there was no association of quantitative genomic load with age, recruitment group, or disease category. In multivariate logistic regression, pneumococcal colonization > 3 log copies/mL was associated with younger age and recruitment group, but not with disease category. CONCLUSIONS: The nasopharyngeal S. pneumoniae colonization patterns of subjects with definite viral infection were very similar to colonization patterns of those with definite bacterial infection or indeterminate pneumonia. Assessment and quantification of nasopharyngeal pneumococcal colonization does not therefore appear useful to discriminate between acute viral and bacterial respiratory disease; consequently, this diagnostic testing is unlikely to reliably determine which children with indeterminate pneumonia have a bacterial etiology and/or require antibiotic treatment.


Assuntos
Nasofaringe/microbiologia , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/etiologia , Streptococcus pneumoniae/isolamento & purificação , Antibacterianos/uso terapêutico , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/tratamento farmacológico , Infecções Bacterianas/epidemiologia , Canadá/epidemiologia , Criança , Pré-Escolar , Estudos de Coortes , Contagem de Colônia Microbiana , Estudos Transversais , Humanos , Lactente , Infecções Respiratórias/tratamento farmacológico , Infecções Respiratórias/epidemiologia , Streptococcus pneumoniae/genética , Viroses/diagnóstico , Viroses/tratamento farmacológico , Viroses/epidemiologia
12.
J Zoo Wildl Med ; 52(2): 638-647, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34130407

RESUMO

To determine the diagnostic value of fecal bacterial enteric pathogen cultures (FBEPC) as part of routine preventive medicine protocols in terrestrial mammals housed in a zoological collection, this study investigated the clinical utility of FBEPC results in context of subsequent clinical actions and how its use was rationalized after adjunct use of fecal cytology as a first-line diagnostic tool. Retrospective results (n = 692) that included a routine FBEPC panel of a commercial diagnostic laboratory, including Aeromonas, Salmonella, Campylobacter, Plesiomonas, Shigella, Yersinia, and Edwardsiella, of 417 mammals were organized into preventive (P; n = 485), diagnostic (D; n = 177), or recheck (R; n = 30) samples; for P and D samples, findings were assigned a "clinical significance factor" of 1 to 5 according to culture results and subsequent clinical actions. A score of 3 or higher indicated positive growth of potentially pathogenic bacterial organisms, of which there were 50 FBEPC (P n = 27, D n = 20, R n = 3). The difference in mean clinical significance factor for P versus D samples was significant. Aeromonas spp. were most frequently isolated (n = 32), followed by Salmonella spp. (n = 8) and Plesiomonas shigelloides (n = 8), then Campylobacter spp. (n = 5). There was no growth of Yersinia enterocolitica, Shigella spp., or Edwardsiella spp. In the absence of clinical evidence of gastrointestinal disease, treatment was initiated in only two cases with isolated Campylobacter spp. Implementation of fecal cytology as an initial step in fecal evaluation resulted in a prompt, substantial reduction in number of ordered FBEPC (mean n = 12/month before and n = 5/month after implementation). The findings in this study suggest that FBEPC for these bacterial species has limited value as a screening tool in preventive medicine protocols for the mammalian orders best represented in this study. The use of fecal cytology led to a more targeted and cost-effective use of FBEPC. Fecal cytology as an initial step in preventative and diagnostic testing protocols is recommended.


Assuntos
Animais de Zoológico , Bactérias/isolamento & purificação , Infecções Bacterianas/veterinária , Fezes/microbiologia , Enteropatias/veterinária , Mamíferos , Animais , Bactérias/classificação , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/microbiologia , Fezes/citologia , Enteropatias/diagnóstico , Enteropatias/microbiologia , Estudos Retrospectivos
13.
Biosens Bioelectron ; 191: 113412, 2021 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-34153636

RESUMO

Phagocytic cells recognize and phagocytose invading microbes for destruction. However, bacterial pathogens can remain hidden at low levels from conventional detection or replicate intracellularly after being phagocytosed by immune cells. Current phagocytosis-detection approaches involve flow cytometry or microscopic search for rare bacteria-internalized phagocytes among large populations of uninfected cells, which poses significant challenges in research and clinical settings. Hence it is imperative to develop a rapid, non-disruptive, and label-free phagocytosis detection approach. Using deformability assays and microscopic imaging, we have demonstrated for the first time that the presence of intracellular bacteria in phagocytic blood cells led to aberrant physical properties. Specifically, human monocytes with internalized bacteria of various species were stiffer and larger compared with uninfected monocytes. Taking advantage of these physical differences, a novel microfluidics-based biosensor platform was developed to passively sort, concentrate and quantify rare monocytes with internalized pathogens (MIP) from uninfected monocyte populations for phagocytosis detection. The clinical utility of the MIP platform was demonstrated by enriching and detecting bacteria-internalized monocytes from spiked human blood samples within 1.5 h. Patient-derived clinical isolates were used to validate the utility of the MIP platform further. This proof-of-concept presents a phagocytosis detection platform that could be used to rapidly diagnose microbial infections, especially in bloodstream infections (BSIs), thereby improving the clinical outcomes for point-of-care management.


Assuntos
Infecções Bacterianas , Técnicas Biossensoriais , Infecções Bacterianas/diagnóstico , Humanos , Monócitos , Fagócitos , Fagocitose
14.
Transfusion ; 61(8): 2414-2420, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34181247

RESUMO

BACKGROUND: In the setting of suspected septic transfusion reactions, bacterial culture of both the transfused patient and the residual blood component is recommended. Primary bacterial contamination can occur at the time of component collection. Clinically insignificant "secondary contamination" can occur during post-transfusion component discard, retrieval for culture, or manipulation of the bag at the time of culture sampling. STUDY DESIGN AND METHODS: This retrospective, multi-center study analyzes positive residual component culture results and companion patient blood cultures from 15 hospitals, 1 blood center, and all cultured transfusion reactions within the province of Quebec, Canada, over a 5-year period. Imputability was assigned as "definite" (concordant growth), "possible" (discordant growth or lack of growth in patient culture), or "unable to assess" (patient not cultured). RESULTS: There were 373 positive component cultures from 360 unique transfusion reactions, with 276 (76.7%) companion patient blood cultures performed, of which 10 (2.8%) yielded the pathogen detected in the positive component. Of these 10 definite pathogens, 7 (2 Staphylococcus aureus, 3 other staphylococci, and 1 Streptococcus pyogenes and 1 Bacillus sp.) were associated with platelet and 3 (Aeromonas veronii, Staphylococcus epidermidis, and Enterococcus faecalis) with RBC transfusions. RBC and plasma components comprised 70% of positive component cultures. DISCUSSION: The process of performing residual component culture is vulnerable to secondary contamination. The significance of microorganisms recovered from component culture cannot be interpreted in isolation. In the context of low prevalence of primary contamination of blood components, the positive predictive value of a positive component culture result is very low.


Assuntos
Bactérias/isolamento & purificação , Infecções Bacterianas/etiologia , Transfusão de Componentes Sanguíneos/efeitos adversos , Segurança do Sangue , Sepse/etiologia , Reação Transfusional/etiologia , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/microbiologia , Hemocultura , Estudos Transversais , Humanos , Estudos Retrospectivos , Sepse/diagnóstico , Sepse/microbiologia , Reação Transfusional/diagnóstico , Reação Transfusional/microbiologia
15.
Auris Nasus Larynx ; 48(6): 1176-1180, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34172351

RESUMO

OBJECTIVE: In Japan, many otolaryngologists provide primary care for patients with coronavirus disease 2019 (COVID-19). The purpose of this study was to analyze the characteristics of otorhinolaryngological findings in order to improve COVID-19 diagnostic systems in a primary care setting. METHODS: A total of 351 patients (mean age, 36.0 ± 15.4 years) diagnosed with COVID-19 by otolaryngologists who belong to the Japan Otorhinolaryngologists Association were included in the study. A web-based questionnaire was used to collect information regarding the timing of positive identification of COVID-19, the route of infection, symptoms, and findings in the tonsils, nasal cavity, pharynx, ear, and neck. A modified Centor score was calculated for cases in which age, symptoms, and tonsil and neck findings were described. RESULTS: Symptoms included fever (56%), olfactory disturbance (46%), and a sore throat (56%). Of the individuals considered, 63% had ordinary rhinoscopic findings, 21% experienced watery rhinorrhea, and 12% had observable mucosal redness. Further, 87% had ordinary tonsillar findings, 13% displayed tonsillar redness, with enlargement and white mucus observe in 2% and 1% of participants, respectively. A total of 193 patients had a calculated Centor score of 3 points in 2%, and scores of the remaining participants were ≤2 points. CONCLUSION: Of all patients considered, 40% had nasal findings and 4% had purulent nasal discharge. In contrast, only 13% of the patients had tonsillar findings, and no patients had Centor scores ≥4 points. Symptom differentiation from that of bacterial infections is difficult. In areas where COVID-19 is prevalent, the disease should be considered in patients presenting with fever, olfactory disturbances, and sore throat with minimal or no clinical findings in the nasal cavity and pharynx.


Assuntos
COVID-19/diagnóstico , Otorrinolaringopatias/diagnóstico , Avaliação de Sintomas , Adulto , Infecções Bacterianas/diagnóstico , COVID-19/complicações , COVID-19/epidemiologia , Diagnóstico Diferencial , Feminino , Pesquisas sobre Serviços de Saúde , Humanos , Japão/epidemiologia , Masculino , Otorrinolaringologistas , Otorrinolaringopatias/epidemiologia , Otorrinolaringopatias/virologia
16.
Methods Mol Biol ; 2344: 151-161, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34115358

RESUMO

Infectious diseases represent a major cause of morbidity and mortality worldwide. Early detection of infections is capital for managing life-threatening cases. So far, traditional diagnostic methods such as microbiological cultures are slow and, sometimes, inaccurate. In the molecular era, high-throughput techniques are essential for providing tools that are able to diagnose in a fast and reliable way, as well as they can be used for monitoring the humoral response of groups of people in a program of epidemiological surveillance when an outbreak occurs, or when a vaccine is being evaluated. Antigen-based protein microarrays are an ideal means for these purposes, as they can carry up to thousands of protein antigens from pathogenic sources and be probed with sera from different human groups (acute or chronic infected people, convalescent, controls). For the diagnosis of bacterial infections, the best antigens are in principle the surface proteins, as they have the highest chances to raise an effective immune response. Here we describe a general protocol for fabricating a glass slide-based protein microarray using recombinant bacterial surface antigens, according to our own expertise in the study of pneumococcal disease. The probing with human sera aims to evaluate differences between diseased and healthy people, in order to discover discriminating antigens that can be used, after appropriate validation, in further easy-to-use formats such as immunostrips.


Assuntos
Antígenos de Bactérias/imunologia , Infecções Bacterianas/imunologia , Análise Serial de Proteínas , Testes Sorológicos , Anticorpos Antibacterianos/imunologia , Infecções Bacterianas/diagnóstico , Vidro/química , Humanos
17.
Eur J Clin Microbiol Infect Dis ; 40(9): 1983-1997, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34109500

RESUMO

SARS-CoV-2 antibody assays are used for epidemiological studies and for the assessment of vaccine responses in highly vulnerable patients. So far, data on cross-reactivity of SARS-CoV-2 antibody assays is limited. Here, we compared four enzyme-linked immunosorbent assays (ELISAs; Vircell SARS-CoV-2 IgM/IgA and IgG, Euroimmun SARS-CoV-2 IgA and IgG) for detection of anti-SARS-CoV-2 antibodies in 207 patients with COVID-19, 178 patients with serological evidence of different bacterial infections, 107 patients with confirmed viral respiratory disease, and 80 controls from the pre-COVID-19 era. In COVID-19 patients, the assays showed highest sensitivity in week 3 (Vircell-IgM/A and Euroimmun-IgA: 78.9% each) and after week 7 (Vircell-IgG: 97.9%; Euroimmun-IgG: 92.1%). The antibody indices were higher in patients with fatal disease. In general, IgM/IgA assays had only limited or no benefit over IgG assays. In patients with non-SARS-CoV-2 respiratory infections, IgG assays were more specific than IgM/IgA assays, and bacterial infections were associated with more false-positive results than viral infections. The specificities in bacterial and viral infections were 68.0 and 81.3% (Vircell-IgM/IgA), 84.8 and 96.3% (Euroimmun-IgA), 97.8 and 86.0% (Vircell-IgG), and 97.8 and 99.1% (Euroimmun-IgG), respectively. Sera from patients positive for antibodies against Mycoplasma pneumoniae, Chlamydia psittaci, and Legionella pneumophila yielded particularly high rates of unspecific false-positive results in the IgM/IgA assays, which was revealed by applying a highly specific flow-cytometric assay using HEK 293 T cells expressing the SARS-CoV-2 spike protein. Positive results obtained with anti-SARS-CoV-2 IgM/IgA ELISAs require careful interpretation, especially if there is evidence for prior bacterial respiratory infections.


Assuntos
Anticorpos Antivirais/sangue , Infecções Bacterianas/diagnóstico , Teste Sorológico para COVID-19/métodos , COVID-19/diagnóstico , Infecções Respiratórias/diagnóstico , Anticorpos Antibacterianos/sangue , Infecções Bacterianas/sangue , COVID-19/sangue , COVID-19/virologia , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Infecções Respiratórias/sangue , SARS-CoV-2/imunologia , Sensibilidade e Especificidade , Glicoproteína da Espícula de Coronavírus/imunologia
18.
Front Immunol ; 12: 631308, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34079538

RESUMO

Febrile patients, suffering from an infection, inflammatory disease or autoimmunity may present with similar or overlapping clinical symptoms, which makes early diagnosis difficult. Therefore, biomarkers are needed to help physicians form a correct diagnosis and initiate the right treatment to improve patient outcomes following first presentation or admittance to hospital. Here, we review the landscape of novel biomarkers and approaches of biomarker discovery. We first discuss the use of current plasma parameters and whole blood biomarkers, including results obtained by RNA profiling and mass spectrometry, to discriminate between bacterial and viral infections. Next we expand upon the use of biomarkers to distinguish between infectious and non-infectious disease. Finally, we discuss the strengths as well as the potential pitfalls of current developments. We conclude that the use of combination tests, using either protein markers or transcriptomic analysis, have advanced considerably and should be further explored to improve current diagnostics regarding febrile infections and inflammation. If proven effective when combined, these biomarker signatures will greatly accelerate early and tailored treatment decisions.


Assuntos
Infecções Bacterianas/diagnóstico , Febre/etiologia , Inflamação/diagnóstico , Viroses/diagnóstico , Infecções Bacterianas/sangue , Infecções Bacterianas/complicações , Biomarcadores/sangue , Diagnóstico Diferencial , Febre/sangue , Febre/microbiologia , Febre/virologia , Perfilação da Expressão Gênica , Humanos , Inflamação/sangue , Inflamação/complicações , Índice de Gravidade de Doença , Viroses/sangue , Viroses/complicações
20.
Int J Mol Sci ; 22(11)2021 May 26.
Artigo em Inglês | MEDLINE | ID: mdl-34073389

RESUMO

The aetiology of Kawasaki disease (KD), an acute inflammatory disorder of childhood, remains unknown despite various triggers of KD having been proposed. Host 'omic profiles offer insights into the host response to infection and inflammation, with the interrogation of multiple 'omic levels in parallel providing a more comprehensive picture. We used differential abundance analysis, pathway analysis, clustering, and classification techniques to explore whether the host response in KD is more similar to the response to bacterial or viral infections at the transcriptomic and proteomic levels through comparison of 'omic profiles from children with KD to those with bacterial and viral infections. Pathways activated in patients with KD included those involved in anti-viral and anti-bacterial responses. Unsupervised clustering showed that the majority of KD patients clustered with bacterial patients on both 'omic levels, whilst application of diagnostic signatures specific for bacterial and viral infections revealed that many transcriptomic KD samples had low probabilities of having bacterial or viral infections, suggesting that KD may be triggered by a different process not typical of either common bacterial or viral infections. Clustering based on the transcriptomic and proteomic responses during KD revealed three clusters of KD patients on both 'omic levels, suggesting heterogeneity within the inflammatory response during KD. The observed heterogeneity may reflect differences in the host response to a common trigger, or variation dependent on different triggers of the condition.


Assuntos
Infecções Bacterianas , Perfilação da Expressão Gênica , Síndrome de Linfonodos Mucocutâneos , Proteômica , Viroses , Adolescente , Infecções Bacterianas/classificação , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/metabolismo , Criança , Pré-Escolar , Biologia Computacional , Feminino , Humanos , Masculino , Síndrome de Linfonodos Mucocutâneos/classificação , Síndrome de Linfonodos Mucocutâneos/diagnóstico , Síndrome de Linfonodos Mucocutâneos/metabolismo , Viroses/classificação , Viroses/diagnóstico , Viroses/metabolismo
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