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1.
J Med Microbiol ; 70(7)2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34259621

RESUMO

Introduction. Contamination of specimens and overuse of broad spectrum antibiotics contribute to false positives and false negatives, respectively. Therefore, useful and applicable biomarkers of bacteremia are still required.Hypothesis/Gap Statement. IL-6 can be used as a serum biomarker to discriminate among bacterial infections and fungal infections in febrile patients with a bloodstream infection.Aim. We aimed to evaluate the diagnostic efficiency of neutrophil/lymphocyte ratio (NLR), procalcitonin (PCT) and interleukin-6 (IL-6) in discriminating Gram-negative (G-) bacteria from Gram-positive (G+) bacteria and fungi in febrile patients.Methodology. A total of 567 patients with fever were evaluated. Serum levels of IL-6, PCT, NLR and CRP were compared among a G- group (n=188), a G+ group (n=168), a fungal group (n=38) and a culture negative group (n=173). Sensitivity, specificity, Yuden's index and area under the Receiver operating characteristic (ROC) curve (AUC) were obtained to analyse the diagnostic abilities of these biomarkers in discriminating bloodstream infection caused by different pathogens.Results. Serum IL-6 and PCT in the G- group increased significantly when compared with both the G+ group and fungal group (P <0.05). AUC of IL-6 (0.767, 95 % CI:0.725-0.805) is higher than AUC of PCT (0.751, 95 % CI:0.708-0.796) in discriminating the G- group from G+ group. When discriminating the G- group from fungal group, the AUC of IL-6 (0.695, 95 % CI:0.651-0.747) with a cut-off value of 464.3 pg ml-1 was also higher than the AUC of PCT (0.630, 95 % CI:0.585-0.688) with a cut-off value of 0.68 ng ml-1. Additionally, AUC of NLR (0.685, 95 % CI:0.646-0.727) in discriminating the fungal group from G+ group at the cut-off value of 9.03, was higher than AUC of IL-6, PCT and CRP.Conclusion. This study suggests that IL-6 could be used as a serum biomarker to discriminate among bacterial infections and fungal infections in febrile patients with a bloodstream infection. In addition, NLR is valuable to discriminate fungal infections from Gram-positive infections in febrile patients with a bloodstream infection.


Assuntos
Biomarcadores/sangue , Febre/sangue , Infecções por Bactérias Gram-Negativas/sangue , Infecções por Bactérias Gram-Positivas/sangue , Micoses/sangue , Adolescente , Adulto , Idoso , Contagem de Células Sanguíneas , Proteína C-Reativa/análise , Calcitonina/sangue , Análise Discriminante , Febre/diagnóstico , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Positivas/diagnóstico , Humanos , Interleucina-6/sangue , Linfócitos/citologia , Masculino , Pessoa de Meia-Idade , Micoses/diagnóstico , Neutrófilos/citologia , Curva ROC , Estudos Retrospectivos , Adulto Jovem
2.
Int J Mol Sci ; 22(11)2021 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-34205910

RESUMO

To overcome the limitations of the Limulus amebocyte lysate (LAL) assay method for the diagnosis of invasive fungal infection, we applied a reaction system combining recombinant ß-glucan binding proteins and a scanning single-molecule counting (SSMC) method. A novel (1→3)-ß-D-glucan recognition protein (S-BGRP) and a (1→6)-ß-glucanase mutant protein were prepared and tested for the binding of (1→6)-branched (1→3)-ß-D-glucan from fungi. S-BGRP and (1→6)-ß-glucanase mutant proteins reacted with ß-glucan from Candida and Aspergillus spp. Although LAL cross-reacted with plant-derived ß-glucans, the new detection system using the SSMC method showed low sensitivity to plant (1→3)-ß-D-glucan, which significantly improved the appearance of false positives, a recognized problem with the LAL method. Measurement of ß-glucan levels by the SSMC method using recombinant ß-glucan-binding proteins may be useful for the diagnosis of fungal infections. This study shows that this detection system could be a new alternative diagnostic method to the LAL method.


Assuntos
Técnicas Biossensoriais , Endotoxinas/isolamento & purificação , Micoses/diagnóstico , beta-Glucanas/isolamento & purificação , Aspergillus/química , Aspergillus/isolamento & purificação , Aspergillus/patogenicidade , Candida/química , Candida/isolamento & purificação , Candida/patogenicidade , Endotoxinas/química , Humanos , Micoses/microbiologia , Imagem Individual de Molécula , beta-Glucanas/química
3.
Int J Mol Sci ; 22(13)2021 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-34281276

RESUMO

Extracellular vesicles (EVs) are membranous, rounded vesicles released by prokaryotic and eukaryotic cells in their normal and pathophysiological states. These vesicles form a network of intercellular communication as they can transfer cell- and function-specific information (lipids, proteins and nucleic acids) to different cells and thus alter their function. Fungi are not an exception; they also release EVs to the extracellular space. The vesicles can also be retained in the periplasm as periplasmic vesicles (PVs) and the cell wall. Such fungal vesicles play various specific roles in the lives of these organisms. They are involved in creating wall architecture and maintaining its integrity, supporting cell isolation and defence against the environment. In the case of pathogenic strains, they might take part in the interactions with the host and affect the infection outcomes. The economic importance of fungi in manufacturing high-quality nutritional and pharmaceutical products and in remediation is considerable. The analysis of fungal EVs opens new horizons for diagnosing fungal infections and developing vaccines against mycoses and novel applications of nanotherapy and sensors in industrial processes.


Assuntos
Vesículas Extracelulares/fisiologia , Fungos/fisiologia , Transporte Biológico Ativo , Vesículas Extracelulares/genética , Vesículas Extracelulares/imunologia , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Fungos/genética , Fungos/patogenicidade , Genes Fúngicos , Interações entre Hospedeiro e Microrganismos/imunologia , Interações entre Hospedeiro e Microrganismos/fisiologia , Humanos , Modelos Biológicos , Micoses/diagnóstico , Micoses/microbiologia , RNA Fúngico/genética , RNA Fúngico/metabolismo
4.
Jt Dis Relat Surg ; 32(2): 556-559, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34145840

RESUMO

Osteomyelitis of the phalanx caused by Candida species are rare. A 49-year-old female patient was admitted to an external center with a splinter injury of the third phalanx of the middle finger of her left hand about 45 days ago. She was referred to our clinic with persistent pain and discharge, despite four-week antibiotherapy. Debridement and curettage were performed and partial excision of the distal phalanx at an appropriate level was done. Her complaints gradually resolved postoperatively with prescribed antibiotics for the pathogen identified as Candida lusitaniae based on the intraoperative cultures. At her three-month follow-up visit, treatment yielded near-excellent results. To the best of our knowledge, this is the first case of osteomyelitis of the distal phalanx caused by Candida lusitaniae in the literature, highlighting the importance of definitive diagnosis and pathogen-specific treatment, rather than empirical treatment, to achieve favorable results with cure.


Assuntos
Falanges dos Dedos da Mão/fisiopatologia , Micoses/diagnóstico , Osteomielite/diagnóstico , Saccharomycetales/isolamento & purificação , Candidíase/diagnóstico , Candidíase/diagnóstico por imagem , Candidíase/microbiologia , Feminino , Humanos , Pessoa de Meia-Idade , Micoses/diagnóstico por imagem , Micoses/microbiologia , Osteomielite/diagnóstico por imagem , Osteomielite/microbiologia , Turquia
5.
BMC Infect Dis ; 21(1): 514, 2021 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-34074256

RESUMO

BACKGROUND: Talaromyces marneffei (TM) bloodstream infection is common in Acquired Immunodeficiency Syndrome (AIDS) patients with extreme immunodeficiency in Southeast Asia and South China, however, clinical case study on TM bloodstream infection is scarce. We retrospectively analyzed the clinical characteristics of TM bloodstream infection in hospitalized AIDS patients and determined the outcomes of hospitalization after diagnosis in our hospital over the past 5 years. METHODS: From January 2015 to July 2020, 87 cases of TM detected by blood culture in patients admitted to our center were collected. The admission complaints, blood cells, biochemistry, CD4 and CD8 cell counts and 1,3-ß-D-glucan (BDG), procalcitonin (PCT), CRP level on the day of blood culture test, and outcomes during hospitalization were analyzed. Logistic regression analysis was performed for the risk factors for poor prognosis (60 cases). Spearman correlation analysis was used to analyze the correlation between peripheral blood cells, albumin and the time required for TM turnaround in blood culture. The difference was statistically significant when the P value was < 0.05. RESULTS: A total of 87 patients were collected, with a median age of 34 years, a median hemoglobin of 94 g/L and CD4 count of 7/µl. The rate of TM bloodstream infection among all in-hospital patients increased from 0.99% in 2015 to 2.09% in 2020(half year). Patients with TM bloodstream infection with CD8 count < 200/µl had a 12.6-fold higher risk of poor prognosis than those with CD8 count > 200/µl (p = 0.04), and those with BDG < 100 pg/mL had a 34.9-fold higher risk of poor prognosis than those with BDG > 100 pg/mL (p = 0.01). CONCLUSIONS: TM bloodstream infection is becoming more common in advanced AIDS patients in endemic areas. For those patients with extremely low CD4 and CD8 cell counts below 200/µl is with an increased risk of poor prognosis.


Assuntos
Infecções Oportunistas Relacionadas com a AIDS/epidemiologia , Fungemia/epidemiologia , Micoses/epidemiologia , Talaromyces/isolamento & purificação , Infecções Oportunistas Relacionadas com a AIDS/diagnóstico , Adulto , China/epidemiologia , Feminino , Fungemia/diagnóstico , Hospitalização , Humanos , Masculino , Pessoa de Meia-Idade , Micoses/diagnóstico , Prognóstico , Estudos Retrospectivos , Fatores de Risco
7.
PLoS Negl Trop Dis ; 15(5): e0009058, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33945531

RESUMO

Talaromyces marneffei is a thermally dimorphic fungus that causes opportunistic systemic mycoses in patients with AIDS or other immunodeficiency syndromes. The purpose of this study was to develop an immunochromatographic strip test (ICT) based on a solid phase sandwich format immunoassay for the detection of T. marneffei antigens in clinical urine specimens. The T. marneffei yeast phase specific monoclonal antibody 4D1 (MAb4D1) conjugated with colloidal gold nanoparticle was used as a specific signal reporter. Galanthus nivalis Agglutinin (GNA) was adsorbed onto nitrocellulose membrane to serve as the test line. Similarly, a control line was created above the test line by immobilization of rabbit anti-mouse IgG. The immobilized GNA served as capturing molecule and as non-immune mediated anti-terminal mannose of T. marneffei antigenic mannoprotein. The MAb4D1-GNA based ICT showed specific binding activity with yeast phase antigen of T. marneffei, and it did not react with other common pathogenic fungal antigens. The limit of detection of this ICT for T. marneffei antigen spiked in normal urine was approximately 0.6 µg/ml. The diagnostic performance of the ICT was validated using 341 urine samples from patents with culture- confirmed T. marneffei infection and from a control group of healthy individuals and patients with other infections in an endemic area. The ICT exhibited 89.47% sensitivity, 100% specificity, and 97.65% accuracy. Our results demonstrate that the urine-based GNA-MAb4D1 based ICT produces a visual result within 30 minutes and that the test is highly specific for the diagnosis of T. marneffei infection. The findings validate the deployment of the ICT for clinical use.


Assuntos
Anticorpos Monoclonais/imunologia , Antígenos de Fungos/urina , Imunoensaio/métodos , Micoses/diagnóstico , Testes Imediatos , Talaromyces/imunologia , Antígenos de Superfície/urina , Ensaio de Imunoadsorção Enzimática/métodos , Coloide de Ouro/química , Humanos , Limite de Detecção , Lectina de Ligação a Manose/imunologia , Lectinas de Ligação a Manose/imunologia , Nanopartículas Metálicas/química , Doenças Negligenciadas/diagnóstico , Doenças Negligenciadas/microbiologia , Lectinas de Plantas/imunologia , Talaromyces/isolamento & purificação
9.
Front Cell Infect Microbiol ; 11: 605589, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33791233

RESUMO

Objective: This study presents a relatively rare case of disseminated Talaromyces marneffei (T. marneffei) infection in an HIV-negative patient. Methods: An 8-month-old girl was hospitalized because of uncontrollable fever and cough for 6 days. Routine laboratory tests, biochemical detection, immunological tests, pathogenic examination, and imaging inspection were performed. Genetic tests of trio whole genome sequencing (Trio-WES), trio copy number sequencing (Trio-CNVseq), and Sanger sequencing were conducted to identify pathogenic variants. In silico analysis of the sequence alignment and structural modeling results was carried out to study the possible pathogenicity of the identified variant. Western blotting was performed to investigate the expression of the identified gene at the protein level. Results: Enhanced CT and MRI scanning demonstrated thymic dysplasia, diffuse pulmonary and liver nodules, and many balloon-like air sacs in both lungs. The white blood cell count, neutrophil count, and neutrophil ratio were normal or elevated. The patient was HIV-negative and bone marrow and blood culture showed T. marneffei infection. Total lymphocyte count, CD3+ T lymphocyte count, CD3+CD4+ T lymphocyte count, CD3+CD8+ T lymphocyte count, and NK cell count decreased, while the number of CD19 positive B cells increased. However, the ratio of CD3+CD4+:CD3+CD8+ T cells increased. Trio-WES identified a homozygous private variant of NM_006509: c.400_c.401insAGC/p.Lys134 delinsLysGln in RELB and Sanger sequencing validated the result. Structural modeling indicated that the variant may be pathogenic. Reverse transcription-polymerase chain reaction and Western blot analysis showed that the expression of RelB in the patient was lower than that in the healthy controls at mRNA and protein levels. Conclusion: This is the first report on disseminated T. marneffei infection in a patient with a homozygous private variant of RELB.


Assuntos
Micoses , Talaromyces , Fator de Transcrição RelB/genética , Feminino , Soronegatividade para HIV , Humanos , Lactente , Pulmão , Micoses/diagnóstico
10.
Vet Clin North Am Exot Anim Pract ; 24(2): 397-418, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33892893

RESUMO

This article is aimed to help the reader to understand better how to diagnose and treat different respiratory diseases in Psittaciformes (parrot-like birds). The article starts from a review of avian respiratory anatomy and physiology, and then moves forward into diagnostic techniques, most common diseases, split in species and anatomic location, and common treatment regimens.


Assuntos
Infecções Bacterianas/veterinária , Doenças das Aves/diagnóstico , Micoses/veterinária , Papagaios/anatomia & histologia , Viroses/veterinária , Animais , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/microbiologia , Infecções Bacterianas/patologia , Doenças das Aves/microbiologia , Doenças das Aves/patologia , Micoses/diagnóstico , Micoses/microbiologia , Micoses/patologia , Papagaios/fisiologia , Viroses/diagnóstico , Viroses/patologia , Viroses/virologia
11.
Clin Infect Dis ; 72(Suppl 2): S109-S113, 2021 03 12.
Artigo em Inglês | MEDLINE | ID: mdl-33709128

RESUMO

The EORTC/MSGERC have revised the definitions for proven, probable, and possible fungal diseases. The tissue diagnosis subcommittee was tasked with determining how and when species can be determined from tissue in the absence of culture. The subcommittee reached a consensus decision that polymerase chain reaction (PCR) from tissue, but not immunohistochemistry or in situ hybridization, can be used for genus or species determination under the new EORTC/MSGERC guidelines, but only when fungal elements are identified by histology. Fungal elements seen in tissue samples by histopathology and identified by PCR followed by sequencing should fulfill the definition of a proven fungal infection, identified to genus/species, even in the absence of culture. This summary discusses the issues that were deliberated by the subcommittee to reach the consensus decision and outlines the criteria a laboratory should follow in order to produce data that meet the EORTC/MSGERC definitions.


Assuntos
Infecções Fúngicas Invasivas , Micoses , Formaldeído , Fungos/genética , Humanos , Infecções Fúngicas Invasivas/diagnóstico , Micoses/diagnóstico , Inclusão em Parafina
12.
Int J Mol Sci ; 22(4)2021 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-33669963

RESUMO

Because Japanese cedar pollen (JCP) contains beta-1,3-d-glucan (BG), there is concern that its lingering presence in the atmosphere, especially during its scattering period, may cause false positives in the factor-G-based Limulus amebocyte lysate (LAL) assay used to test for deep mycosis (i.e., G-test). Hence, we examined whether the LAL assay would react positively with substances contained in JCP by using the G-test to measure JCP particles and extracts. BG was purified from the JCP extract on a BG-specific affinity column, and the percentage extractability was measured using three different BG-specific quantitative methods. The G-test detected 0.4 pg BG in a single JCP particle and 10 fg from a single particle in the extract. The percentage extractability of JCP-derived BG was not significantly different among the three quantitative methods. As the JCP particles should technically have been removed during serum separation, they should be less likely to be a direct false-positive factor. However, given that the LAL-assay-positive substances in the JCP extract were not distinguishable by the three BG-specific quantitative methods, we conclude that they may cause the background to rise. Therefore, in Japan false positives arising from JCP contamination should be considered when testing patients for deep mycosis.


Assuntos
Cryptomeria/imunologia , Micoses/diagnóstico , Pólen/imunologia , Reações Falso-Positivas , Concentração de Íons de Hidrogênio , Lectinas Tipo C/metabolismo , beta-Glucanas/metabolismo
13.
BMC Infect Dis ; 21(1): 287, 2021 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-33743629

RESUMO

BACKGROUND: Talaromyces marneffei (T. marneffei) is an opportunistic pathogen that infects immunodeficient children. The aim of the study is to determine the clinical features and peripheral immune state of Talaromyces marneffei (T. marneffei) infections in children for early detection and diagnosis. METHODS: We retrospectively reviewed 21 pediatric patients who were diagnosed with T. marneffei infections and were followed up in the Guangzhou Women and Children's Medical Center from January 2010 to January 2020. For each patient, we collected and analyzed clinical characteristics, peripheral immunological results, genetic tests, complications and prognosis. RESULTS: Common clinical features of the patients included fever (20/21, 95.24%), cough (17/21, 80.95%) and hepatomegaly (17/21, 80.95%). Severe complications included septic shock (12/21, 57.14%), hemophagocytic lymphohistiocytosis (HLH) (11/21, 52.38%), acute respiratory distress syndrome (ARDS) (10/21, 47.62%), multiple organ dysfunction syndrome (MODS) (9/21, 42.86%), and disseminated intravascular coagulation (DIC) (7/21, 33.33%). Eleven children (11/21, 52.38%) eventually died of T. marneffei infections. All patients were HIV negative. Seven cases revealed reduced antibody levels, especially IgG. Higher levels of IgE were detected in 9 cases with an obvious increase in two patients. Ten patients showed decreased complement C3 levels, some of whom had low C4 levels. Three patients displayed decreased absolute T lymphocyte counts, including the CD 4+ and CD 8+ subsets. A reduction in NK cells was present in most patients. No patient had positive nitro blue tetrazolium (NBT) test results. Nine patients were screened for common genetic mutations. Of the cases, one case had no disease-specific gene mutation. Four children had confirmed hyperimmunoglobulin M syndrome (HIGM) with CD40LG variation, one case had severe combined immunodeficiency (SCID), and one case had hyper-IgE syndrome (HIES). One patient was identified as having a heterozygous mutation in STAT3 gene; however, he showed no typical clinical manifestations of HIES at his age. One patient had a mutated COPA gene with uncertain pathogenic potential. Another patient was diagnosed with HIES that depended on her clinical features and the National Institutes of Health (NIH) scoring system. CONCLUSIONS: T. marneffei infections in HIV-negative children induced severe systemic complications and poor prognosis. Children with T. marneffei infections commonly exhibited abnormal immunoglobulin levels in peripheral blood, particularly decreased IgG or increased IgE levels, further suggesting possible underlying PIDs in these patients.


Assuntos
Anticorpos Antifúngicos/sangue , Micoses/imunologia , Criança , China , Diagnóstico Precoce , Feminino , Soronegatividade para HIV , Humanos , Masculino , Micoses/diagnóstico , Estudos Retrospectivos , Talaromyces
14.
Lancet Infect Dis ; 21(8): e246-e257, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-33606997

RESUMO

With increasing numbers of patients needing intensive care or who are immunosuppressed, infections caused by moulds other than Aspergillus spp or Mucorales are increasing. Although antifungal prophylaxis has shown effectiveness in preventing many invasive fungal infections, selective pressure has caused an increase of breakthrough infections caused by Fusarium, Lomentospora, and Scedosporium species, as well as by dematiaceous moulds, Rasamsonia, Schizophyllum, Scopulariopsis, Paecilomyces, Penicillium, Talaromyces and Purpureocillium species. Guidance on the complex multidisciplinary management of infections caused by these pathogens has the potential to improve prognosis. Management routes depend on the availability of diagnostic and therapeutic options. The present recommendations are part of the One World-One Guideline initiative to incorporate regional differences in the epidemiology and management of rare mould infections. Experts from 24 countries contributed their knowledge and analysed published evidence on the diagnosis and treatment of rare mould infections. This consensus document intends to provide practical guidance in clinical decision making by engaging physicians and scientists involved in various aspects of clinical management. Moreover, we identify areas of uncertainty and constraints in optimising this management.


Assuntos
Micoses/diagnóstico , Micoses/tratamento farmacológico , Animais , Gerenciamento Clínico , Fungos/efeitos dos fármacos , Fungos/genética , Fungos/isolamento & purificação , Fungos/fisiologia , Humanos , Micologia , Micoses/microbiologia , Guias de Prática Clínica como Assunto , Sociedades Médicas
15.
Clin Lab ; 67(2)2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-33616342

RESUMO

BACKGROUND: Rapid and accurate diagnosis of HIV-positive patients with Talaromyces marneffei (T. marneffei) infections remains challenging. A 60-year-old woman came to our inpatient department presenting with hematuria, abdominal pain, and diarrhea for one week. The patient had a past medical history of Acquired Immune Deficiency Syndrome (AIDS). The patient's stool was watery and the color of soy sauce. The patient was without fever, cough, and skin lesions. METHODS: The blood routine was performed with a Mindray BC-6900 hematology analyzer. RESULTS: Blood routine showed leukocytosis with neutrophilia and basophils and the WBC/DIFF scattergram showed a cluster of neutrophils connected with a monocyte and lymphocyte cluster and an additional cluster of immature granulocytes and heterotypic lymphocytes or primitive cells. Surprisingly, the peripheral blood film evaluation revealed small round-to-ovoid yeast cells within the cytoplasm of neutrophils. A T. marneffei infection was suspected and anti-fungal therapy was initiated. The patient's diarrhea improved after treatment with amphotericin B for two days. A second blood routine showed a normal number of leukocytes and basophils and a diminished cluster of immature granulocytes and heterotypic lymphocytes or primitive cells. After one week, blood cultures had grown T. marneffei. CONCLUSIONS: The WBC/DIFF scattergram obtained from a Mindray BC-6900 analyzer provided significant hints to enhance diagnosis of T. marneffei when combined with results of a peripheral blood smear.


Assuntos
Soropositividade para HIV , Micoses , Talaromyces , Feminino , Humanos , Pessoa de Meia-Idade , Micoses/diagnóstico , Micoses/tratamento farmacológico
17.
Infect Control Hosp Epidemiol ; 42(1): 84-88, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32703320
18.
Arch Pathol Lab Med ; 145(2): 145-167, 2021 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-32886738

RESUMO

CONTEXT.­: Point-of-care testing (POCT) is inherently spatial, that is, performed where needed, and intrinsically temporal, because it accelerates decision-making. POCT efficiency and effectiveness have the potential to facilitate antimicrobial resistance (AMR) detection, decrease risks of coinfections for critically ill patients with coronavirus infectious disease 2019 (COVID-19), and improve the cost-effectiveness of health care. OBJECTIVES.­: To assess AMR identification by using POCT, describe the United States AMR Diagnostic Challenge, and improve global standards of care for infectious diseases. DATA SOURCES.­: PubMed, World Wide Web, and other sources were searched for papers focusing on AMR and POCT. EndNote X9.1 (Clarivate Analytics) consolidated abstracts, URLs, and PDFs representing approximately 500 articles were assessed for relevance. Panelist insights at Tri•Con 2020 in San Francisco and finalist POC technologies competing for a US $20,000,000 AMR prize are summarized. CONCLUSIONS.­: Coinfections represent high risks for COVID-19 patients. POCT potentially will help target specific pathogens, refine choices for antimicrobial drugs, and prevent excess morbidity and mortality. POC assays that identify patterns of pathogen resistance can help tell us how infected individuals spread AMR, where geospatial hotspots are located, when delays cause death, and how to deploy preventative resources. Shared AMR data "clouds" could help reduce critical care burden during pandemics and optimize therapeutic options, similar to use of antibiograms in individual hospitals. Multidisciplinary health care personnel should learn the principles and practice of POCT, so they can meet needs with rapid diagnostic testing. The stakes are high. Antimicrobial resistance is projected to cause millions of deaths annually and cumulative financial loses in the trillions by 2050.


Assuntos
COVID-19/microbiologia , Coinfecção/microbiologia , Farmacorresistência Bacteriana , Farmacorresistência Fúngica , Testes de Sensibilidade Microbiana/métodos , Sistemas Automatizados de Assistência Junto ao Leito , Distinções e Prêmios , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/microbiologia , COVID-19/diagnóstico , COVID-19/mortalidade , Coinfecção/diagnóstico , Humanos , Testes de Sensibilidade Microbiana/normas , Micoses/diagnóstico , Micoses/microbiologia , Sistemas Automatizados de Assistência Junto ao Leito/normas , Análise Espacial , Estados Unidos/epidemiologia
19.
Clin Infect Dis ; 72(8): 1386-1391, 2021 04 26.
Artigo em Inglês | MEDLINE | ID: mdl-32155243

RESUMO

BACKGROUND: The toxigenic mold Stachybotrys has controversially been linked to idiopathic pulmonary hemorrhage and "sick building syndrome." However, there are no previous clinical records of invasive stachybotryosis. METHODS: Sinus biopsy specimens from a 23-year-old male with refractory acute lymphocytic leukemia were obtained at 3 different time points during the patient's hospitalization (139 days) and examined by histopathology and immunohistochemistry (IHC). Antifungal susceptibility testing and fungal speciation using multilocus sequence typing were performed. RESULTS: Hemorrhage, fungal germination, and hyphal growth were observed in the first sinus biopsy tissues. Areas with fungal growth tested positive for Stachybotrys by IHC. Fungal isolates were genotyped and identified as Stachybotrys chlorohalonata. The patient was cured from Stachybotrys sinusitis following sinus surgery and antifungal treatment. While a subsequent second sinus biopsy and a bronchoscopy showed no signs of fungal infection, a later, third sinus biopsy tested positive for Aspergillus calidoustus, a rare human pathogen. CONCLUSIONS: Here, we report the first case of invasive S. chlorohalonata sinusitis that was surgically and medically cured but followed by invasive A. calidoustus sinusitis in the setting of refractory leukemia. Our findings emphasize the risk for unusual fungal infections in severely immunocompromised patients.


Assuntos
Micoses , Sinusite , Stachybotrys , Adulto , Aspergillus , Humanos , Masculino , Micoses/diagnóstico , Micoses/tratamento farmacológico , Sinusite/diagnóstico , Adulto Jovem
20.
J Med Microbiol ; 70(2)2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33252325

RESUMO

Introduction. Histopathological examination (HPE) of tissue helps in the diagnosis of invasive fungal infections (IFIs) but cannot identify the fungus to the genus/species levelGap Statement Available protocols for the molecular identification of fungi from formalin-fixed and paraffin-embedded (FFPE) tissues have limitations in terms of extraction and target selection, and standardisation.Aim. Development of sequence-based fungal identification protocol after extraction of DNA from formalin-fixed and paraffin-embedded (FFPE) tissues.Methodology. A total of 63 FFPE tissues from histopathology proven IFI cases were used to standardize the DNA extraction (commercial QIAamp kit-based extraction and conventional phenol-chloroform-isoamyl alcohol [PCI] method) and sequence-based fungal identification protocols. The PCR targeted different ribosomal DNA (rDNA) regions including complete internal transcribed spacer (ITS1-5.8S-ITS2), separate ITS1 and ITS2, 18S and D1/D2 of 28S regions. Semi-nested PCR targeting Mucorales-specific 18S rDNA region was performed in tissues having aseptate hyphae. The optimized ITS1-PCR protocol was evaluated in 119 FFPE tissues containing septate hyphae or yeast, and Mucorales-specific semi-nested PCR in 126 FFPE tissues containing aseptate hyphae.Results. The DNA yield by conventional PCI method was significantly higher (P<0.0001) than commercial kit, though the quality of DNA was similar by both protocols. The test accuracy was best while using ITS1 (61.9 %) as the target compared to 7.9, 29.9 and 22.2 % on targeting ITS1-5.8S-ITS2, ITS2, the D1/D2 region of 28S, respectively. The test accuracies of ITS1-PCR in tissues containing septate hyphae, aseptate hyphae and yeasts were 75.5, 18.7 and 100 %, respectively. The amplification (targeting ITS1 region) improved by increasing the thickness of tissue section (up to 50 µm) used for DNA extraction. ITS1-PCR protocol could amplify fungal DNA in 76 (63.8 %) tissues and Mucorales-specific semi-nested PCR in 86 (68.3 %) tissues.Conclusion. Conventional PCI-based DNA extraction from thick tissue (50 µm) may be used until optimal commercial fungal DNA extraction kit is developed. Subsequent ITS1-PCR for septate fungi and yeast, and semi-nested PCR targeting 18S rDNA for Mucorales are recommended to identify the fungus in FFPE tissues.


Assuntos
DNA Fúngico/genética , Fungos/classificação , Fungos/genética , Tipagem Molecular/métodos , Técnicas de Tipagem Micológica/métodos , DNA Espaçador Ribossômico/genética , Formaldeído , Humanos , Técnicas de Diagnóstico Molecular , Micoses/diagnóstico , Micoses/microbiologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Inclusão em Parafina , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 18S/genética , RNA Ribossômico 28S/genética
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