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SUMMARYThis guidance presents recommendations for clinical microbiology laboratories for processing respiratory samples from people with cystic fibrosis (pwCF). Appropriate processing of respiratory samples is crucial to detect bacterial and fungal pathogens, guide treatment, monitor the epidemiology of cystic fibrosis (CF) pathogens, and assess therapeutic interventions. Thanks to CF transmembrane conductance regulator modulator therapy, the health of pwCF has improved, but as a result, fewer pwCF spontaneously expectorate sputum. Thus, the collection of sputum samples has decreased, while the collection of other types of respiratory samples such as oropharyngeal and bronchoalveolar lavage samples has increased. To optimize the detection of microorganisms, including Pseudomonas aeruginosa, Staphylococcus aureus, Haemophilus influenzae, and Burkholderia cepacia complex; other less common non-lactose fermenting Gram-negative bacilli, e.g., Stenotrophomonas maltophilia, Inquilinus, Achromobacter, Ralstonia, and Pandoraea species; and yeasts and filamentous fungi, non-selective and selective culture media are recommended for all types of respiratory samples, including samples obtained from pwCF after lung transplantation. There are no consensus recommendations for laboratory practices to detect, characterize, and report small colony variants (SCVs) of S. aureus, although studies are ongoing to address the potential clinical impact of SCVs. Accurate identification of less common Gram-negative bacilli, e.g., S. maltophilia, Inquilinus, Achromobacter, Ralstonia, and Pandoraea species, as well as yeasts and filamentous fungi, is recommended to understand their epidemiology and clinical importance in pwCF. However, conventional biochemical tests and automated platforms may not accurately identify CF pathogens. MALDI-TOF MS provides excellent genus-level identification, but databases may lack representation of CF pathogens to the species-level. Thus, DNA sequence analysis should be routinely available to laboratories for selected clinical circumstances. Antimicrobial susceptibility testing (AST) is not recommended for every routine surveillance culture obtained from pwCF, although selective AST may be helpful, e.g., for unusual pathogens or exacerbations unresponsive to initial therapy. While this guidance reflects current care paradigms for pwCF, recommendations will continue to evolve as CF research expands the evidence base for laboratory practices.
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Solid organ transplant recipients (SOTRs) frequently receive adjunctive glucocorticoid therapy (AGT) for Pneumocystis jirovecii pneumonia (PJP). This multicenter cohort of SOTRs with PJP admitted to 20 transplant centers in Canada, the United States, Europe, and Australia, was examined for whether AGT was associated with a lower rate of all-cause intensive care unit (ICU) admission, 90-day death, or a composite outcome (ICU admission or death). Of 172 SOTRs with PJP (median [IQR] age: 60 (51.5-67.0) years; 58 female [33.7%]), the ICU admission and death rates were 43.4%, and 20.8%, respectively. AGT was not associated with a reduced risk of ICU admission (adjusted odds ratio [aOR] [95% CI]: 0.49 [0.21-1.12]), death (aOR [95% CI]: 0.80 [0.30-2.17]), or the composite outcome (aOR [95% CI]: 0.97 [0.71-1.31]) in the propensity score-adjusted analysis. AGT was not significantly associated with at least 1 unit of the respiratory portion of the Sequential Organ Failure Assessment score improvement by day 5 (12/37 [32.4%] vs 39/111 [35.1%]; P = .78). We did not observe significant associations between AGT and ICU admission or death in SOTRs with PJP. Our findings should prompt a reevaluation of routine AGT administration in posttransplant PJP treatment and highlight the need for interventional studies.
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Transplante de Órgãos , Pneumocystis carinii , Pneumonia por Pneumocystis , Feminino , Humanos , Pessoa de Meia-Idade , Europa (Continente) , Glucocorticoides/uso terapêutico , Transplante de Órgãos/efeitos adversos , Pneumonia por Pneumocystis/tratamento farmacológico , Pneumonia por Pneumocystis/etiologia , Estudos Retrospectivos , Transplantados , Masculino , IdosoRESUMO
PURPOSE: Although 5% povidone-iodine (PVP-I) is frequently used as an ocular antiseptic agent, there is a lack of consensus regarding the effects of PVP-I concentration, storage after opening, and compounded preparation on PVP-I antisepsis. We performed a series ofâ¯in-vitro experiments to determine the impact of these factors on PVP-I's inhibition of common causes of post-procedural eye infection. METHODS: Inhibition of microorganism growth was measured in-vitroâ¯as a function of active PVP-I exposure time. In control experiments, PVP-I was inactivated before microorganism exposure. Tested PVP-I solutions varied in concentration (0.6%, 5%, or 10%), length of storage after opening (0, 7, or 30 days), and preparation (commercial vs.compounded from stock PI solution). Tested pathogens includedâ¯S. epidermidis, S. viridans, P. aeruginosa, methicillin-resistant S. aureus, methicillin-sensitive S. aureus, andâ¯C. albicans. RESULTS: PVP-I solutions inhibited all bacterial growth by 3 min and fungal growth by 15 s. Compared to 5% PVP-I, the 0.6% PVP-I was less effective in inhibiting S. viridans growth (200 ± 0 colonies vs. 7 ± 8 at 30 s, P = 0.0004; 183 ± 21 vs. 0 ± 0 at 1 min, P = 0.018), but more effective in inhibiting P. aeruginosa (30 ± 20 vs. 200 ± 0 at 15 s, P = 0.019). Compared to commercial and newly-opened PVP-I solutions, compounded preparations and solutions stored for 7 or 30 days after bottle opening either preserved or improved antiseptic efficacy against tested microorganisms. CONCLUSIONS: Concentration of PVP-I solution affects antiseptic efficacy within 1 min of exposure, but all solutions performed equivalently at 3 min. In contrast to results of prior studies investigating dilute PVP-I, the 0.6% PVP-I did not demonstrate a uniformly equivalent or superior anti-septic effect. Compounded preparation and storage length after bottle opening did not decrease PVP-I antiseptic activity.
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Histoplasmosis, the most common endemic mycosis in North America, presents in a myriad of ways, spanning the spectrum from self-limiting pneumonia to progressive disseminated histoplasmosis (PDH). Toward better describing contemporary histoplasmosis syndromes, risks, and outcomes, this single-center retrospective cohort study was performed (2009-2019). The population who developed PDH was similar to that with other forms of histoplasmosis (OFH) except for higher rates of preexisting immunocompromising conditions (91.3% vs. 40%, P < .001) and a trend toward receiving more chronic immunosuppression (65.2% vs. 33.3%, P = .054) compared to those with OFH. Diagnosis was most frequently achieved by urinary or serum antigen positivity. People with PDH more frequently tested positive compared to those with OFH, but negative tests did not rule out histoplasmosis. Median time to diagnosis was prolonged among people with both PDH and OFH (32 vs. 31 days, respectively). Following diagnosis, people with PDH received more liposomal amphotericin (78.3% vs. 20%, P < .001). Subsequent survival at 90 and 365 days and treatment response were similar in both groups. Patients with PDH were more often hospitalized (95.7% vs. 60%, P = .006); however, once admitted, there were no differences in hospital length of stay or intensive care unit admission rate. The challenges of diagnosing histoplasmosis based on clinical presentation alone highlight the need for heightened awareness of these entities especially given the recent reports on expanded endemicity and delays in diagnosis.
Histoplasmosis is the most common endemic mycosis in North America. This article summarizes the clinical features, risk factors, and outcomes in patients who developed disseminated disease compared to more localized forms of histoplasmosis.
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Histoplasmose , Humanos , Histoplasmose/diagnóstico , Histoplasmose/tratamento farmacológico , Histoplasmose/epidemiologia , Histoplasmose/veterinária , Estudos Retrospectivos , Hospedeiro Imunocomprometido , Terapia de Imunossupressão/veterinária , HospitaisRESUMO
BACKGROUND: Coronavirus disease 2019 (COVID-19)-associated pulmonary aspergillosis (CAPA) occurs in critically ill patients with COVID-19. Risks and outcomes remain poorly understood. METHODS: A retrospective cohort study of mechanically ventilated adult patients with COVID-19 admitted to 5 Johns Hopkins hospitals was conducted between March and August 2020. CAPA was defined using composite clinical criteria. Fine and Gray competing risks regression was used to analyze clinical outcomes and, multilevel mixed-effects ordinal logistic regression was used to compare longitudinal disease severity scores. RESULTS: In the cohort of 396 people, 39 met criteria for CAPA. Patients with CAPA were more likely than those without CAPA to have underlying pulmonary vascular disease (41% vs 21.6%, respectively; Pâ =â .01), liver disease (35.9% vs 18.2%; Pâ =â .02), coagulopathy (51.3% vs 33.1%; Pâ =â .03), solid tumors (25.6% vs 10.9%; Pâ =â .02), multiple myeloma (5.1% vs 0.3%; Pâ =â .03), and corticosteroid exposure during the index admission (66.7% vs 42.6%; Pâ =â .005), and had lower body mass indexes (median, 26.6 vs 29.9 [calculated as weight in kilograms divided by height in meters squared]; Pâ =â .04). Patients with CAPA had worse outcomes, as measured by ordinal severity of disease scores, requiring longer time to improvement (adjusted odds ratio, 1.081.091.1; Pâ <â .001), and advancing in severity almost twice as quickly (subhazard ratio, 1.31.82.5; Pâ <â .001). They were intubated twice as long as those without CAPA (subhazard ratio, 0.40.50.6; Pâ <â .001) and had longer hospital stays (median [interquartile range], 41.1 [20.5-72.4) vs 18.5 [10.7-31.8] days; Pâ <â .001). CONCLUSION: CAPA is associated with poor outcomes. Attention to preventive measures (screening and/or prophylaxis) is warranted in people with high risk of CAPA.
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COVID-19 , Aspergilose Pulmonar Invasiva , Aspergilose Pulmonar , Adulto , Humanos , Aspergilose Pulmonar Invasiva/diagnóstico , Aspergilose Pulmonar Invasiva/epidemiologia , Respiração Artificial/efeitos adversos , Estudos Retrospectivos , SARS-CoV-2RESUMO
PURPOSE: Increased ophthalmology-specific risk of novel coronavirus 2019 (SARS-CoV-2) transmission is well-established, increasing the fear of infection and causing associated decreased rates of procedures known to save vision. However, the potential transmission from exposure to clinic instrumentation is unknown, including which additional pathogens may be spreading in this context. This study seeks to fill this gap by characterizing the microbiota of instrumentation in ophthalmology clinics during the COVID-19 pandemic and identifying potential sources of pathogenic spread encountered by patients and healthcare workers. METHODS: Thirty-three samples were captured using standard cultures and media. Ten positive and negative controls were used to confirm proper technique. Descriptive statistics were calculated for all samples. Samples were collected from the retina (N = 17), glaucoma (N = 6), cornea (N = 6), and resident (N = 4) clinics with rigorous disinfection standards at a tertiary academic medical center. Standard media cultures and/or polymerase chain reaction (PCR) was performed for each sample. RESULTS: From 33 samples, more than half (17/33, 51.5%) yielded bacterial growth. Using two different molecular methods, three samples (3/33, 9%) tested positive for SARS-CoV-2 (cycle thresholds 36.48, 37.14, and 37.83). There was no significant difference in bacterial growth (95% confidence interval [95% CI]: - 0.644-0.358, p = 0.076) among different clinics (retina, glaucoma, cornea, resident). Staphylococcus (S.) epidermidis grew most frequently (12/35, 34%), followed by S. capitis (7/35, 20%), Micrococcus luteus (2/35, 5.7%), Corynebacterium tuberculostearicum (2/35, 5.7%), and Cutibacterium ([C.], Propionibacterium) acnes (2/35, 5.7%). C. acnes growth was more frequent with imaging device forehead rests (2/7, 28.6%) than other surfaces (0/26, 0%, 95% CI: 0.019-0.619, p = 0.040). No samples isolated fungus or adenovirus. CONCLUSIONS: Most samples across subspecialty clinic instrumentation grew bacteria, and several tested positive for SARS-CoV-2. Many isolated pathogens have been implicated in causing infections such as endophthalmitis, conjunctivitis, uveitis, and keratitis. The clinical implications of the ophthalmology microbiome for transmitting nosocomial infections warrant optimization of disinfection practices, strategies for mitigating spread, and additional study beyond the pandemic.
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COVID-19 , Glaucoma , Microbiota , Oftalmologia , COVID-19/epidemiologia , Humanos , Pandemias , SARS-CoV-2RESUMO
Aspergillosis complicating severe influenza infection has been increasingly detected worldwide. Recently, coronavirus disease-associated pulmonary aspergillosis (CAPA) has been detected through rapid reports, primarily from centers in Europe. We provide a case series of CAPA, adding 20 cases to the literature, with review of pathophysiology, diagnosis, and outcomes. The syndromes of pulmonary aspergillosis complicating severe viral infections are distinct from classic invasive aspergillosis, which is recognized most frequently in persons with neutropenia and in other immunocompromised persons. Combined with severe viral infection, aspergillosis comprises a constellation of airway-invasive and angio-invasive disease and results in risks associated with poor airway fungus clearance and killing, including virus- or inflammation-associated epithelial damage, systemic immunosuppression, and underlying lung disease. Radiologic abnormalities can vary, reflecting different pathologies. Prospective studies reporting poor outcomes in CAPA patients underscore the urgent need for strategies to improve diagnosis, prevention, and therapy.
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COVID-19/complicações , Aspergilose Pulmonar/complicações , SARS-CoV-2 , Humanos , Fatores de RiscoRESUMO
Establishing the diagnosis of invasive mold infections (IMI) in immunocompromised children is challenging due to nonspecific clinical presentations and the limited sensitivity of traditional culture-based methods. Rapid non-culture-based diagnostics such as the 1,3-beta-d-glucan and galactomannan assays have emerged as promising adjuncts to conventional diagnostic tests in adults. Available data suggest that 1,3-beta-d-glucan has limited accuracy in the pediatric population and is not recommended to be used for the diagnosis of IMI in children. On the other hand, the diagnostic performance of the serum and bronchoalveolar lavage galactomannan in immunocompromised children is comparable to results observed in adults and can be used as a screening tool in children at high risk of developing invasive aspergillosis (IA) who are not receiving mold-active antifungal prophylaxis and as a diagnostic tool in symptomatic children suspected of having IA. Herein, we summarize the available evidence for the use of these rapid non-culture-based diagnostics in immunocompromised children. We also summarize potential causes of false positivity for the 1,3-beta-d-glucan and galactomannan assays.
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Aspergilose , beta-Glucanas , Adulto , Aspergilose/diagnóstico , Criança , Galactose/análogos & derivados , Glucanos , Humanos , Hospedeiro Imunocomprometido , Mananas , Sensibilidade e EspecificidadeRESUMO
Fungal infections are a rising threat to our immunocompromised patient population, as well as other nonimmunocompromised patients with various medical conditions. However, little progress has been made in the past decade to improve fungal diagnostics. To jointly address this diagnostic challenge, the Fungal Diagnostics Laboratory Consortium (FDLC) was recently created. The FDLC consists of 26 laboratories from the United States and Canada that routinely provide fungal diagnostic services for patient care. A survey of fungal diagnostic capacity among the 26 members of the FDLC was recently completed, identifying the following diagnostic gaps: lack of molecular detection of mucormycosis; lack of an optimal diagnostic algorithm incorporating fungal biomarkers and molecular tools for early and accurate diagnosis of Pneumocystis pneumonia, aspergillosis, candidemia, and endemic mycoses; lack of a standardized molecular approach to identify fungal pathogens directly in formalin-fixed paraffin-embedded tissues; lack of robust databases to enhance mold identification with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; suboptimal diagnostic approaches for mold blood cultures, tissue culture processing for Mucorales, and fungal respiratory cultures for cystic fibrosis patients; inadequate capacity for fungal point-of-care testing to detect and identify new, emerging or underrecognized, rare, or uncommon fungal pathogens; and performance of antifungal susceptibility testing. In this commentary, the FDLC delineates the most pressing unmet diagnostic needs and provides expert opinion on how to fulfill them. Most importantly, the FDLC provides a robust laboratory network to tackle these diagnostic gaps and ultimately to improve and enhance the clinical laboratory's capability to rapidly and accurately diagnose fungal infections.
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Laboratórios , Mucorales , Canadá , Técnicas de Laboratório Clínico , Prova Pericial , HumanosRESUMO
Scedosporium aurantiacum is one of the emergent opportunistic fungal pathogens among immunocompromised hosts. Colonization of S. aurantiacum can also occur in patients with underlying lung diseases such as cystic fibrosis. S. aurantiacum is highly resistant to multiple antifungal agents, and management of the infected patients can be very challenging compared to those infected with other species of Scedosporium. Clinical cases have been geographically restricted mostly in Australia with a small number of cases identified in Europe and Japan. Although clinical isolates of S. aurantiacum from the USA have been included in several research studies, no clinical case of S. aurantiacum infection from the USA has been described. We report a case of S. aurantiacum infection acquired in the SA. Awareness of S. aurantiacum among healthcare providers and the species-level identification for Scedosporium are critically important for appropriate selection of antifungal agents and improvement of treatment outcomes.
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Infecções Fúngicas Invasivas , Scedosporium , Antifúngicos/uso terapêutico , Humanos , Estados UnidosRESUMO
BACKGROUND: Cryptococcosis is increasingly recognized in people without human immunodeficiency virus (HIV). METHODS: A multicenter, prospective cohort study was performed in 25 US centers. Consenting patients were prospectively followed for ≤2 years. Neurological morbidities were assessed with longitudinal event depiction and functional scores (Montreal Cognitive Assessment [MoCA]). Risks of death were analyzed using Cox regression. RESULTS: One hundred forty-five subjects were enrolled. Most were male (95; 65.5%) and had immunosuppression (120; 82.8%), including solid organ transplant (SOT; 33.8%), autoimmunity (15.9%), and hematologic malignancies (11.7%). Disease involved the central nervous system (CNS) in 71 subjects (49%). Fever was uncommon, documented in 40 (27.8%) subjects, and absence was associated with diagnostic delay (mean: 48.2 vs 16.5 days; P = .007). Abnormal MoCA scores (<26) were predictive of CNS disease; low scores (<22) were associated with poor long-term cognition. Longitudinal event depiction demonstrated frequent complications in people with CNS disease; 25 subjects (35.2%) required >1 lumbar puncture and 8 (11.3%) required ventriculostomies. In multivariable models, older age (>60 years) was associated with higher risks of death (hazard ratio [HR], 2.14; 95% confidence interval [CI], 1.05-4.38; P = .036), and lower risks were noted with underlying hematologic malignancy (HR, 0.29; 95% CI, 0.09-0.98; P = .05) and prior SOT (HR, 0.153; 95% CI, 0.05-0.44; P = .001). CONCLUSIONS: Despite aggressive antifungal therapies, outcomes of CNS cryptococcosis in people without HIV are characterized by substantial long-term neurological sequelae. Studies are needed to understand mechanism(s) of cognitive decline and to enable better treatment algorithms.
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Criptococose , Diagnóstico Tardio , Idoso , Estudos de Coortes , Criptococose/epidemiologia , HIV , Humanos , Estudos Longitudinais , Masculino , Estudos Prospectivos , Estados Unidos/epidemiologiaRESUMO
Lomentospora prolificans is an opportunistic fungal pathogen with low susceptibility to current antifungal drugs. Here, we tested the in vitro susceptibility of 8 drugs against 42 clinical L. prolificans isolates. All isolates showed high MICs to voriconazole (MIC90>16 µg/ml), itraconazole (MIC90>16 µg/ml), posaconazole (MIC90>16 µg/ml), isavuconazole (MIC90>16 µg/ml), amphotericin B (MIC90>16 µg/ml), and terbinafine (MIC90>64 µg/ml) and high minimum effective concentrations (MECs) to micafungin (MEC90>8 µg/ml), with the exception of miltefosine showing an MIC90 value of 4 µg/ml. We examined six different in vitro drug combinations and found that the combination of voriconazole and terbinafine achieved the most synergistic effort against L. prolificans We then annotated the L. prolificans whole genome and located its Cyp51 and Fks1 genes. We completely sequenced the two genes to determine if any mutation would be related to azole and echinocandin resistance in L. prolificans We found no amino acid changes in Cyp51 protein and no tandem repeats in the 5' upstream region of the Cyp51 gene. However, we identified three intrinsic amino acid residues (G138S, M220I, and T289A) in the Cyp51 protein that were linked to azole resistance. Likewise, two intrinsic amino acid residues (F639Y, W695F) that have reported to confer echinocandin resistance were found in Fks1 hot spot regions. In addition, three new amino acid alterations (D440A, S634R, and H1245R) were found outside Fks1 hot spot regions, and their contributions to echinocandin resistance need future investigation. Overall, our findings support the notion that L. prolificans is intrinsically resistant to azoles and echinocandins.
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Antifúngicos , Scedosporium , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Aspergillus fumigatus , Azóis , Farmacorresistência Fúngica/genética , Equinocandinas/farmacologia , Proteínas Fúngicas/genética , Testes de Sensibilidade MicrobianaRESUMO
Routine identification of fungal pathogens from positive blood cultures by culture-based methods can be time-consuming, delaying treatment with appropriate antifungal agents. The GenMark Dx ePlex investigational use only blood culture identification fungal pathogen panel (BCID-FP) rapidly detects 15 fungal targets simultaneously in blood culture samples positive for fungi by Gram staining. We aimed to determine the performance of the BCID-FP in a multicenter clinical study. Blood culture samples collected at 10 United States sites and tested with BCID-FP at 4 sites were compared to the standard-of-care microbiological and biochemical techniques, fluorescence in situ hybridization using peptide nucleic acid probes (PNA-FISH) and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Discrepant results were analyzed by bi-directional PCR/sequencing of residual blood culture samples. A total of 866 clinical samples, 120 retrospectively and 21 prospectively collected, along with 725 contrived samples were evaluated. Sensitivity and specificity of detection of Candida species (C. albicans, C. auris, C. dubliniensis, C. famata, C. glabrata, C. guilliermondii, C. kefyr, C. krusei, C. lusitaniae, C. parapsilosis, and C. tropicalis) ranged from 97.1 to 100% and 99.8 to 100%, respectively. For the other organism targets, sensitivity and specificity were as follows: 100% each for Cryptococcus neoformans and C. gattii, 98.6% and 100% for Fusarium spp., and 96.2% and 99.9% for Rhodotorula spp., respectively. In 4 of the 141 clinical samples, the BCID-FP panel correctly identified an additional Candida species, undetected by standard-of-care methods. The BCID-FP panel offers a faster turnaround time for identification of fungal pathogens in positive blood cultures that may allow for earlier antifungal interventions and includes C. auris, a highly multidrug-resistant fungus.
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Hemocultura , Microfluídica , Fungos/genética , Humanos , Hibridização in Situ Fluorescente , Reação em Cadeia da Polimerase , Estudos RetrospectivosAssuntos
Infecções Fúngicas Invasivas , Meningite Fúngica , beta-Glucanas , Humanos , Glucanos , Sistema Nervoso Central , Meningite Fúngica/diagnóstico , Meningite Fúngica/tratamento farmacológico , Infecções Fúngicas Invasivas/diagnóstico , Infecções Fúngicas Invasivas/tratamento farmacológico , Líquido CefalorraquidianoAssuntos
Hemocultura , Reação em Cadeia da Polimerase Multiplex , Rhodotorula , Humanos , Reação em Cadeia da Polimerase Multiplex/métodos , Hemocultura/métodos , Rhodotorula/genética , Rhodotorula/isolamento & purificação , Reações Falso-Positivas , Técnicas de Diagnóstico Molecular/métodos , Fungemia/diagnóstico , Fungemia/microbiologiaRESUMO
BACKGROUND: Pulmonary Cryptococcosis (PC) is diagnosed with increasing incidence in recent years, but it does not commonly involve the pleural space. Here, we report a HIV-negative case with advanced stage IIIB non-small cell lung cancer (NSCLC) treated with radiation therapy presented with dyspnea, a new PET-positive lung mass and bilateral pleural effusion suspecting progressive cancer. However, the patient has been diagnosed as pulmonary cryptococcal infection and successfully treated with oral fluconazole therapy. CASE PRESENTATION: A 77-year-old male with advanced stage non-small cell lung cancer treated with combined chemo-radiation therapy who presented with progressive dyspnea, a new PET-positive left lower lobe lung mass and bilateral pleural effusions. Initial diagnostic thoracentesis and bronchoscopy yielded no cancer, but instead found yeast forms consistent with cryptococcal organisms in the transbronchial biopsies of the left lower lobe lung mass. Subsequent to this, the previously collected pleural fluid culture showed growth of Cryptococcus neoformans. The same sample of pleural effusion was tested and was found to be positive for crytococcal antigen (CrAg) by a lateral flow assay (LFA). The patient has been treated with oral fluconazole therapy resulting in gradual resolution of the nodular infiltrates. CONCLUSION: PC should be considered in immunosuppressed cancer patients. Additionally, concomitant pleural involvement in pulmonary cryptococcal infections may occur. The incidence of false positive 18FDG-PET scans in granulomatous infections and the use of CrAg testing in pleural fluid to aid in diagnosis are reviewed.
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Criptococose/diagnóstico por imagem , Criptococose/microbiologia , Pneumopatias Fúngicas/diagnóstico por imagem , Pneumopatias Fúngicas/microbiologia , Derrame Pleural/microbiologia , Administração Oral , Idoso , Antifúngicos/administração & dosagem , Antifúngicos/uso terapêutico , Carcinoma Pulmonar de Células não Pequenas/complicações , Carcinoma Pulmonar de Células não Pequenas/radioterapia , Criptococose/tratamento farmacológico , Cryptococcus neoformans/patogenicidade , Fluconazol/administração & dosagem , Fluconazol/uso terapêutico , Humanos , Hospedeiro Imunocomprometido , Pneumopatias Fúngicas/tratamento farmacológico , Neoplasias Pulmonares/complicações , Neoplasias Pulmonares/radioterapia , Masculino , Derrame Pleural/diagnóstico por imagem , Derrame Pleural/etiologia , Tomografia por Emissão de PósitronsRESUMO
Colletotrichum is an important fungal plant pathogen, yet an uncommon cause of human disease. Herein we report a case of invasive, cutaneous infection in a stem cell transplant recipient due to Colletotrichum species, with accompanying review of the literature. The infection was successfully treated with a combination of liposomal amphotericin B and voriconazole. Multilocus phylogenetic analysis revealed that the distinct isolate belongs to Colletotrichum siamense, a member of the Colletotrichum gloeosporioides species complex not previously described as a human pathogen. Colletotrichum infection remains in the differential for skin lesions in the immune compromised host.
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Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Infecções Fúngicas Invasivas/etiologia , Pele/microbiologia , Adulto , Idoso , Pré-Escolar , Colletotrichum/genética , Feminino , Humanos , Hospedeiro Imunocomprometido , Infecções Fúngicas Invasivas/microbiologia , Masculino , Pessoa de Meia-Idade , Pele/patologiaRESUMO
Clinical tissues are prepared for histological analysis and long-term storage via formalin fixation and paraffin embedding (FFPE). The FFPE process results in fragmentation and chemical modification of RNA, rendering it less suitable for analysis by techniques that rely on reverse transcription (RT) such as RT-qPCR and RNA-Seq. Here we describe a broadly applicable technique called 'Ligation in situ Hybridization' ('LISH'), which is an alternative methodology for the analysis of FFPE RNA. LISH utilizes the T4 RNA Ligase 2 to efficiently join adjacent chimeric RNA-DNA probe pairs hybridized in situ on fixed RNA target sequences. Subsequent treatment with RNase H releases RNA-templated ligation products into solution for downstream analysis. We demonstrate several unique advantages of LISH-based assays using patient-derived FFPE tissue. These include >100-plex capability, compatibility with common histochemical stains and suitability for analysis of decade-old materials and exceedingly small microdissected tissue fragments. High-throughput DNA sequencing modalities, including single molecule sequencing, can be used to analyze ligation products from complex panels of LISH probes ('LISH-seq'), which can be amplified efficiently and with negligible bias. LISH analysis of FFPE RNA is a novel methodology with broad applications that range from multiplexed gene expression analysis to the sensitive detection of infectious organisms.
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Hibridização In Situ/métodos , Inclusão em Parafina/métodos , RNA/genética , Fixação de Tecidos/métodos , Perfilação da Expressão Gênica/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Microscopia de Fluorescência , RNA/análise , RNA/metabolismo , RNA Ligase (ATP)/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Ribonuclease H/metabolismo , Proteínas Virais/metabolismoRESUMO
Among clinician-ordered vaginal cultures positive for Candida albicans , 30% exhibited fluconazole resistance. Resistance did not reliably predict future susceptibility. Prospective studies to verify associations with demographic and clinical factors as well as to correlate in vitro resistance with treatment response and longitudinal resistance patterns are needed.