RESUMEN
Several false positive low serum cryptococcal antigen (SCrAg) reports by lateral flow assay (LFA) were identified in late 2016 at our tertiary care hospital. After the recall and correction of the problem in the reagent, we studied the significance of SCrAg LFA ≤ 1:10 from January 2017 to October 2023. Of 20 patients with 31 samples of SCrAg LFA ≤ 1:10, 14 patients (70%) were classified as true positives, four (20%) were indeterminate, and only two (10%) patients were false positives. If a new SCrAg LFA ≤ 1:10 is detected, it should be repeated, and additional workup should be pursued.
We studied the significance of low serum cryptococcal antigen (SCrAg) titer lateral flow assay (LFA) ≤ 1:10 from January 2017 to October 2023. Of 20 patients with SCrAg LFA ≤ 1:10, only two patients (10%) were false positives. If a new SCrAg ≤ 1:10 is detected, it should be repeated, and additional workup should be done.
Asunto(s)
Antígenos Fúngicos , Criptococosis , Cryptococcus , Centros de Atención Terciaria , Humanos , Antígenos Fúngicos/sangre , Antígenos Fúngicos/inmunología , Criptococosis/diagnóstico , Criptococosis/sangre , Masculino , Femenino , Cryptococcus/inmunología , Persona de Mediana Edad , Reacciones Falso Positivas , Adulto , Anciano , Estudios RetrospectivosRESUMEN
An adult female rhesus macaque presented during routine annual physical examination for evaluation of a 2.5-cm diameter superficial ulcerated dermal lesion that was subsequently diagnosed as a systemic fungal infection caused by Cryptococcus gattii. Cryptococcus gattii is one of several basidiomycetic yeasts responsible for pulmonary, neurologic, and disseminated infections in humans and animals. This report describes the diagnosis, management, and clinical resolution of a C. gattii infection in an immunocompetent 5-year-old female rhesus macaque.
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Antifúngicos , Criptococosis , Cryptococcus gattii , Macaca mulatta , Enfermedades de los Monos , Animales , Cryptococcus gattii/aislamiento & purificación , Criptococosis/veterinaria , Criptococosis/tratamiento farmacológico , Criptococosis/diagnóstico , Criptococosis/microbiología , Femenino , Enfermedades de los Monos/microbiología , Enfermedades de los Monos/diagnóstico , Enfermedades de los Monos/tratamiento farmacológico , Antifúngicos/uso terapéutico , InmunocompetenciaRESUMEN
OBJECTIVE: Cryptococcosis predominantly presents as a meningoencephalitis in Thailand. Early and expeditious diagnosis is essential for reducing both mortality and morbidity associated with cryptococcal meningitis. We aim to define and establish the diagnostic performances between the benchmark commercially available diagnostic kit (CrAg® LFA) and the large-scale prototype of an inexpensive in-house immunochromatographic test (ICT) based on monoclonal antibody (MAb) 18B7. METHODS: We have developed the large-scale prototype for the rapid detection of cryptococcal polysaccharide antigens by utilizing a single antibody sandwich ICT format employing MAb 18B7, which is highly specific to Cryptococcus neoformans glucuronoxylomannan (GXM) antigens. An in-house MAb18B7 ICT was manufactured in accordance with industry standards under the control of the International Organization for Standardization (ISO) 13485. RESULTS: The diagnostic sensitivity, specificity, and accuracy for the in-house MAb 18B7 ICT were 99.10%, 97.61%, and 97.83%, respectively. The agreement kappa (κ) coefficient was 0.968 based on the retrospective evaluation of 580 specimens from patients living in northern Thailand with clinically suspected cryptococcosis. CONCLUSION: The data suggest that this in-house MAb 18B7 ICT will be highly beneficial for addressing the issue of cryptococcal infection in Thailand. Moreover, it is anticipated that this inexpensive ICT can play a pivotal role in various global strategies aimed at eradicating cryptococcal meningitis among individuals living with HIV by 2030.
Asunto(s)
Anticuerpos Monoclonales , Antígenos Fúngicos , Cromatografía de Afinidad , Criptococosis , Cryptococcus neoformans , Sensibilidad y Especificidad , Humanos , Tailandia , Anticuerpos Monoclonales/inmunología , Cromatografía de Afinidad/métodos , Criptococosis/diagnóstico , Cryptococcus neoformans/inmunología , Cryptococcus neoformans/aislamiento & purificación , Antígenos Fúngicos/análisis , Antígenos Fúngicos/inmunología , Estudios Retrospectivos , Anticuerpos Antifúngicos/sangre , Polisacáridos/análisis , Polisacáridos/inmunología , Masculino , Femenino , Adulto , Pruebas Diagnósticas de Rutina/métodos , Persona de Mediana Edad , Anciano , Adulto JovenRESUMEN
Cryptococcosis is a systemic zoonotic disease that is challenging to diagnose based on clinical findings in cats and dogs due to the nonspecific nature of its clinical presentation. This case report aims to document the first confirmed case of disseminated cryptococcosis caused by Cryptococcus neoformans in a dog in Turkey and to highlight the potential link between natural disasters such as earthquakes and the emergence of zoonotic diseases in domestic animals. A two-and-a-half-year-old spayed female Cocker Spaniel presented with increased respiratory sounds, skin lesions, facial swelling and enlarged lymph nodes. These symptoms appear to be a complication of Demodex infestation due to the stress experienced by the dog following exposure to a severe earthquake. Diagnostic procedures including cytologic examination, fungal culture and DNA sequence analysis, which confirmed the infection was caused by C. neoformans. Due to the delay in the correct diagnosis of the disease, which, contrary to common data, started as an allergic reaction on the skin and was later diagnosed as a Demodex infestation, the dog died of severe respiratory failure during the treatment with itraconazole. The case highlights the critical role of veterinary emergency and critical care in the diagnosis and management of zoonotic diseases post-natural disasters. It also highlights the need for increased awareness and preparedness among veterinary professionals to address animal health challenges following such events.
Asunto(s)
Criptococosis , Enfermedades de los Perros , Zoonosis , Animales , Perros , Femenino , Criptococosis/diagnóstico , Criptococosis/veterinaria , Criptococosis/tratamiento farmacológico , Criptococosis/microbiología , Enfermedades de los Perros/microbiología , Enfermedades de los Perros/diagnóstico , Enfermedades de los Perros/parasitología , Enfermedades de los Perros/tratamiento farmacológico , Zoonosis/microbiología , Zoonosis/diagnóstico , Zoonosis/parasitología , Turquía , Cryptococcus neoformans/aislamiento & purificación , Desastres , Humanos , Resultado Fatal , Antifúngicos/uso terapéuticoRESUMEN
Cryptococcal infection of central nervous system commonly involves meningitis or meningoencephalitis, but rarely mimics inflammatory myelitis. We present short segment myelitis as a dominant manifestation caused by Cryptococcus neoformans in a patient with nephrotic syndrome under immunosuppressive therapy. This case report highlights Cryptococcus neoformans as a potential etiological factor for short segment myelitis in immunocompromised hosts.
Asunto(s)
Criptococosis , Cryptococcus neoformans , Mielitis , Humanos , Mielitis/microbiología , Mielitis/tratamiento farmacológico , Mielitis/diagnóstico , Criptococosis/tratamiento farmacológico , Criptococosis/microbiología , Criptococosis/diagnóstico , Cryptococcus neoformans/aislamiento & purificación , Masculino , Huésped Inmunocomprometido , Persona de Mediana Edad , Síndrome Nefrótico/complicaciones , Inmunosupresores/uso terapéutico , Imagen por Resonancia MagnéticaRESUMEN
Pulmonary cryptococcosis (PC) is a common opportunistic fungal infection caused by Cryptococcus neoformans or Cryptococcus gattii. PC primarily invades the respiratory system, followed by the central nervous system. Few clinical reports have examined the coexistence of PC and lung cancer. This study reports the case of a 54-year-old immunocompetent PC patient with lung adenocarcinoma. Chest CT revealed multiple nodules in the right lung, with the largest nodule located in the dorsal segment of the right lower lobe. 18 FFDG positron emission tomography-computed tomography (PET-CT) revealed elevated glucose metabolism in the dorsal segment of the right lower lobe, which suggested lung cancer. The metabolism level of the nodule in the basal segment of the right lower lobe and the anterior segment of the right upper lobe was not abnormally increased, but the possibility of a malignant tumour could not be excluded. The pulmonary nodules in the dorsal segment and the basal segment of the right lower lobe were simultaneously resected via video-assisted thoracic surgery (VATS), and the final histopathology revealed primary lung adenocarcinoma and pulmonary cryptococcal infection, respectively. After surgery, antifungal treatment was administered for 3 months. Over the 3-year follow-up, contrast-enhanced computed tomography (CT) revealed no recurrence of either disease. This case study highlights the possibility of dualism in the diagnosis of multiple pulmonary nodules on chest CT, such as the coexistence of lung cancer and PC. Surgical resection is recommended for micronodules that are not easy to diagnose via needle biopsy; in addition, early diagnosis and treatment are helpful for ensuring a good prognosis. This paper reports the clinical diagnosis and treatment of one patient with pulmonary cryptococcal infection of the right lung complicated with lung adenocarcinoma, including 3 years of follow-up, providing a reference for clinical practice.
Asunto(s)
Adenocarcinoma del Pulmón , Criptococosis , Enfermedades Pulmonares Fúngicas , Neoplasias Pulmonares , Tomografía Computarizada por Tomografía de Emisión de Positrones , Humanos , Persona de Mediana Edad , Adenocarcinoma del Pulmón/complicaciones , Adenocarcinoma del Pulmón/diagnóstico , Adenocarcinoma del Pulmón/patología , Adenocarcinoma del Pulmón/cirugía , Antifúngicos/administración & dosificación , Criptococosis/complicaciones , Criptococosis/diagnóstico , Criptococosis/patología , Criptococosis/terapia , Pulmón/diagnóstico por imagen , Pulmón/microbiología , Pulmón/patología , Pulmón/cirugía , Enfermedades Pulmonares Fúngicas/complicaciones , Enfermedades Pulmonares Fúngicas/diagnóstico , Enfermedades Pulmonares Fúngicas/patología , Enfermedades Pulmonares Fúngicas/terapia , Neoplasias Pulmonares/complicaciones , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/cirugía , Cirugía Torácica Asistida por Video , Tomografía Computarizada por Rayos XRESUMEN
BACKGROUND: Cryptococcosis is an infectious disease caused by encapsulated heterobasidiomycete yeasts. As an opportunistic pathogen, cryptococcal inhalation infection is the most common. While Primary cutaneous cryptococcosis is extremely uncommon. CASE PRESENTATION: A 61-year-old woman with a history of rheumatoid arthritis on long-term prednisone developed a red plaque on her left thigh. Despite initial antibiotic treatment, the erythema worsened, leading to rupture and fever. Microbiological analysis of the lesion's secretion revealed Candida albicans, Pseudomonas aeruginosa, and methicillin-resistant Staphylococcus epidermidis. Skin biopsy showed thick-walled spores, and culture confirmed primary cutaneous infection with Cryptococcus neoformans. Histopathological stains were positive, and mass spectrometry identified serotype A of the pathogen. The patient was treated with oral fluconazole and topical nystatin, resulting in significant improvement and near-complete healing of the skin lesion within 2.5 months. CONCLUSIONS: Primary cutaneous cryptococcosis was a primary skin infection exclusively located on the skin. It has no typical clinical manifestation of cutaneous infection of Cryptococcus, and culture and histopathology remain the gold standard for diagnosing. The recommended medication for Primary cutaneous cryptococcosis is fluconazole. When patients at risk for opportunistic infections develop skin ulcers that are unresponsive to antibiotic, the possibility of primary cutaneous cryptococcosis needs to be considered.
Asunto(s)
Antifúngicos , Criptococosis , Cryptococcus neoformans , Fluconazol , Humanos , Femenino , Persona de Mediana Edad , Cryptococcus neoformans/aislamiento & purificación , Cryptococcus neoformans/efectos de los fármacos , Criptococosis/tratamiento farmacológico , Criptococosis/microbiología , Criptococosis/diagnóstico , Criptococosis/patología , Antifúngicos/uso terapéutico , Fluconazol/uso terapéutico , Dermatomicosis/tratamiento farmacológico , Dermatomicosis/microbiología , Dermatomicosis/diagnóstico , Dermatomicosis/patología , Piel/patología , Piel/microbiología , Resultado del Tratamiento , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/complicacionesRESUMEN
BACKGROUND Cryptococcosis is an opportunistic fungal infection that typically occurs in patients with compromised immune systems, primarily affecting the respiratory and central nervous systems. However, cryptococcal osteomyelitis is a rare manifestation of cryptococcal infection, characterized by nonspecific clinical features. Here, we present a case of vertebral cryptococcal osteomyelitis in a middle-aged woman and discuss diagnostic approaches. CASE REPORT A 56-year-old woman presented with lower back pain and limited mobility, without fever, and with a history of pulmonary tuberculosis. Physical examination revealed enlarged lymph nodes and tenderness in the thoracic vertebrae. A computed tomography-guided biopsy confirmed granulomatous inflammation caused by Cryptococcus, with abundant 10 µm spherical microbial spores. After 4 weeks of treatment with amphotericin B and fluconazole, symptoms and lesions improved. Upon discharge, the patient was prescribed oral fluconazole. Follow-up examinations showed a stable condition and a negative serum cryptococcal capsular polysaccharide antigen test. CONCLUSIONS Given the rarity and lack of specificity of clinical features of cryptococcal spondylitis, clinicians encountering similar presentations should consider tuberculous spondylitis and spinal tumors as differential diagnoses. Additionally, tissue biopsy of the affected vertebral bodies should be performed early to establish the type of vertebral infection, aiding in diagnosis, treatment, and prognosis.
Asunto(s)
Criptococosis , Osteomielitis , Tuberculosis de la Columna Vertebral , Humanos , Femenino , Persona de Mediana Edad , Osteomielitis/diagnóstico , Osteomielitis/microbiología , Osteomielitis/tratamiento farmacológico , Criptococosis/diagnóstico , Criptococosis/tratamiento farmacológico , Diagnóstico Diferencial , Tuberculosis de la Columna Vertebral/diagnóstico , Vértebras Torácicas , Antifúngicos/uso terapéutico , Fluconazol/uso terapéutico , Tomografía Computarizada por Rayos XRESUMEN
BACKGROUND: Cryptococcosis is a life-threatening disease caused by Cryptococcus neoformans or C. gattii. Neutralizing autoantibodies (auto-Abs) against granulocyte-macrophage colony-stimulating factor (GM-CSF) in otherwise healthy adults with cryptococcal meningitis have been described since 2013. We searched for neutralizing auto-Abs in sera collected from Colombian patients with non-HIV-associated cryptococcosis in a retrospective national cohort from 1997 to 2016. METHODS: We reviewed clinical and laboratory records and assessed the presence of neutralizing auto-Abs against GM-CSF in 30 HIV negative adults with cryptococcosis (13 caused by C. gattii and 17 caused by C. neoformans). RESULTS: We detected neutralizing auto-Abs against GM-CSF in the sera of 10 out of 13 (77%) patients infected with C. gattii and one out of 17 (6%) patients infected with C. neoformans. CONCLUSIONS: We report eleven Colombian patients diagnosed with cryptococcosis who had auto-Abs that neutralize GM-CSF. Among these patients, ten were infected with C. gattii and only one with C. neoformans.
Asunto(s)
Anticuerpos Neutralizantes , Autoanticuerpos , Criptococosis , Cryptococcus gattii , Cryptococcus neoformans , Factor Estimulante de Colonias de Granulocitos y Macrófagos , Humanos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Autoanticuerpos/sangre , Autoanticuerpos/inmunología , Masculino , Colombia , Femenino , Adulto , Cryptococcus gattii/inmunología , Persona de Mediana Edad , Cryptococcus neoformans/inmunología , Criptococosis/inmunología , Criptococosis/diagnóstico , Anticuerpos Neutralizantes/sangre , Anticuerpos Neutralizantes/inmunología , Estudios Retrospectivos , Seronegatividad para VIH/inmunología , Adulto Joven , AncianoRESUMEN
Cryptococcosis is a fungal infection burdened by a high case-fatality rate in immunocompromised patients. Once limited to human immunodeficiency virus (HIV)-infected patients, the epidemiology of cryptococcosis has evolved in recent years and new risk factors have emerged. It is therefore essential to identify these risk factors in order to improve prevention and therapeutic efficacy. We conducted a retrospective observational study including all cases of cryptococcosis between January 2016 and December 2022, diagnosed at the University Hospital of Marseille. During the study period 15 cases of cryptococcosis were diagnosed. Six patients were HIV-infected. Nine patients had one or more comorbidities including liver cirrhosis, type 2 diabetes mellitus, primary immunodeficiency disorder, chronic lymphocytic leukemia and solid organ transplantation. Ten patients had central nervous system cryptococcosis, four had pulmonary cryptococcosis and one patient had extra-pulmonary disseminated cryptococcosis. Of the three patients with liver cirrhosis, two patients died with a post-mortem diagnosis. Our data suggest that emerging risk factors are probably underestimated by clinicians. It emphasizes the need for cryptococcal antigenemia as part of syndromic investigation of any unexplained fever or neurological symptoms in an at-risk patient. Early diagnosis and treatment are essential for patient's survival.
Asunto(s)
Criptococosis , Hospitales Universitarios , Huésped Inmunocomprometido , Humanos , Criptococosis/epidemiología , Criptococosis/diagnóstico , Masculino , Femenino , Estudios Retrospectivos , Hospitales Universitarios/estadística & datos numéricos , Persona de Mediana Edad , Adulto , Anciano , Factores de Riesgo , Francia/epidemiología , Infecciones por VIH/complicaciones , Infecciones por VIH/epidemiología , Cirrosis Hepática/complicaciones , Cirrosis Hepática/epidemiologíaRESUMEN
BACKGROUND Cryptococcosis is an invasive fungal infection caused by Cryptococcus species complex. C. neoformans is one of the pathogenic species within the genus. C. neoformans infections often present as an opportunistic infection in severely immunocompromised individuals. Infection of the pericardium in the setting of liver failure is uncommon. We present a case of cryptococcal pericarditis in a patient with liver failure. CASE REPORT A 47-year-old man with a past medical history of psoriatic arthritis, and alcohol use disorder presented to the emergency department with a 2-week history of progressively worsening generalized weakness, malaise, and yellowish skin changes. Physical examination revealed scleral icterus, jaundiced skin, and ascites. Initial laboratory workup revealed thrombocytopenia, transaminitis (aspartate transaminase (AST) level of 502 IU/L, alanine transaminase (ALT) level of 82 IU/L), hyperbilirubinemia (total bilirubin of 15.7 mg/dL), International Nationalized Ratio (INR) of 3.6, and lactic acidosis (lactic acid of 11.7 mmol/L). The patient developed encephalopathy and acute hypoxic respiratory failure requiring intubation. A bedside point-of-care cardiac ultrasound, performed following intubation, revealed a pericardial effusion without signs of tamponade. This finding was later confirmed by a formal transthoracic echocardiogram. Percutaneous pericardiocentesis was performed, and the pericardial fluid culture revealed the presence of C. neoformans. Human immunodeficiency virus (HIV) tests were negative. The patient received antifungal therapy. Due to his poor prognosis, he was transitioned to comfort care and eventually died. CONCLUSIONS This case report describes an unusual presentation of acute liver failure complicated by cryptococcal pericarditis, emphasizing the importance of considering atypical fungal infections in such patients.
Asunto(s)
Criptococosis , Pericarditis , Humanos , Masculino , Persona de Mediana Edad , Criptococosis/diagnóstico , Criptococosis/complicaciones , Pericarditis/microbiología , Pericarditis/diagnóstico , Fallo Hepático/etiología , Cryptococcus neoformans/aislamiento & purificación , Antifúngicos/uso terapéutico , Huésped Inmunocomprometido , Resultado FatalRESUMEN
A 2-year-old Norwegian Forest cat was presented for evaluation of bilateral purulent nasal discharge and stertorous breathing. A computed tomography (CT) scan of the head revealed an intranasal mass of the left nasal cavity extending behind the tube openings and completely obstructing the nasopharynx. Rhinoscopy confirmed a pinkish, shiny mass. CT scan showed both compartments of the right middle ear filled with abnormal soft tissue attenuating material. There was no change in the bony outline of the middle ear. In the endoscopic examination, after endoscopically assisted tympanocentesis, this material in the accessible dorsolateral compartment proved to be classic polypous tissue in addition to highly viscous glue-like secretions. A secondary otitis media due to a drainage disorder was suspected.Using an endoscopic-interventional approach through the nostril, the nasopharyngeal mass was removed for histopathological examination, in order to restore the nasal airway, and to allow tube drainage. In contrast to cats with classical malignant nasal cavity masses, the cat showed several attachment points of the mass and multiple undulating elevations bilaterally in the nasopharyngeal mucosa.Cytological and histopathological examination identified the mass as a fungal granuloma in the context of a cryptococcus infection only rarely observed in Germany. Molecular genetic analysis confirmed an infection with Cryptococcus neoformans var. grubii.A single intranasal and nasopharyngeal endoscopic debridement resulted in a significant improvement of the clinical signs and a complete healing of the right middle ear (including the tympanic membrane) within 14 days, but not in a complete cure of the disease. The cat was therefore treated with oral itraconazole solution for several weeks.The case report shows that nasal cryptococcosis can also affect cats in Germany. Rhinoscopy reveals a nasopharyngeal mass with multiple attachment points, which is unusual for a neoplasia. In addition to the recommended removal of the mass, oral administration of systemic antimycotics is strongly advised.
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Enfermedades de los Gatos , Criptococosis , Animales , Gatos , Enfermedades de los Gatos/diagnóstico , Enfermedades de los Gatos/microbiología , Enfermedades de los Gatos/patología , Criptococosis/veterinaria , Criptococosis/diagnóstico , Criptococosis/microbiología , Criptococosis/patología , Criptococosis/tratamiento farmacológico , Diagnóstico Diferencial , Neoplasias Nasofaríngeas/veterinaria , Neoplasias Nasofaríngeas/diagnóstico , Alemania , Tomografía Computarizada por Rayos X/veterinaria , Enfermedades Nasofaríngeas/veterinaria , Enfermedades Nasofaríngeas/diagnóstico , Enfermedades Nasofaríngeas/microbiología , Enfermedades Nasofaríngeas/patologíaRESUMEN
ABSTRACT: This article reports an elderly male patient with nodules and ulcers on the face and behind the left ear after trauma. Primary cutaneous cryptococcosis was confirmed using pathological biopsy, special staining, tissue culture, and fungal sequencing. The patient received a therapeutic intervention involving the administration of the antifungal agent itraconazole. Substantial amelioration of cutaneous manifestations was observed after a 3-month course of treatment. After an elapsed interval, the patient was diagnosed with esophageal tumor. Moreover, the literature on 33 patients with primary cutaneous cryptococcosis published in the past 10 years was also reviewed.
Asunto(s)
Antifúngicos , Criptococosis , Dermatomicosis , Humanos , Criptococosis/tratamiento farmacológico , Criptococosis/patología , Criptococosis/microbiología , Criptococosis/diagnóstico , Masculino , Antifúngicos/uso terapéutico , Dermatomicosis/tratamiento farmacológico , Dermatomicosis/microbiología , Dermatomicosis/patología , Dermatomicosis/diagnóstico , Anciano , Itraconazol/uso terapéutico , Neoplasias Esofágicas/patología , Neoplasias Esofágicas/microbiología , Neoplasias Esofágicas/tratamiento farmacológico , Resultado del Tratamiento , Biopsia , Cryptococcus neoformans/aislamiento & purificaciónRESUMEN
Cryptococcus neoformans is a fungal pathogen that causes cryptococcosis primarily in immunocompromised patients, such as those with HIV/AIDS. One survival mechanism of C. neoformans during infection is melanin production, which catalyzed by laccase and protects fungal cells against immune attack. Hence, the comparative assessment of laccase activity is useful for characterizing cryptococcal strains. We serendipitously observed that culturing C. neoformans with food coloring resulted in degradation of some dyes with phenolic structures. Consequently, we investigated the color changes for the food dyes metabolized by C. neoformans laccase and by using this effect explored the development of a colorimetric assay to measure laccase activity. We developed several versions of a food dye-based colorimetric laccase assay that can be used to compare the relative laccase activities between different C. neoformans strains. We found that phenolic color degradation was glucose-dependent, which may reflect changes in the reduction properties of the media. Our food color-based colorimetric assay has several advantages, including lower cost, irreversibility, and not requiring constant monitoring , over the commonly used 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay for determining laccase activity. This method has potential applications to bioremediation of water pollutants in addition to its use in determining laccase virulence factor expression.IMPORTANCECryptococcus neoformans is present in the environment, and while infection is common, disease occurs mostly in immunocompromised individuals. C. neoformans infection in the lungs results in symptoms like pneumonia, and consequently, cryptococcal meningitis occurs if the fungal infection spreads to the brain. The laccase enzyme catalyzes the melanization reaction that serves as a virulence factor for C. neoformans. Developing a simple and less costly assay to determine the laccase activity in C. neoformans strains can be useful for a variety of procedures ranging from studying the relative virulence of cryptococci to environmental pollution studies.
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Cryptococcus neoformans , Colorantes de Alimentos , Lacasa , Humanos , Colorimetría/métodos , Criptococosis/microbiología , Criptococosis/diagnóstico , Cryptococcus neoformans/enzimología , Colorantes de Alimentos/metabolismo , Proteínas Fúngicas/metabolismo , Lacasa/metabolismo , Fenoles/metabolismoRESUMEN
BACKGROUND: Cryptococcosis is a global invasive mycosis associated with significant morbidity and mortality. Cryptococcal antigen (CrAg) testing from serum and cerebrospinal fluid (CSF) has been regarded as a gold standard for early diagnosis. This study aimed to develop and validate a rapid and sensitive sandwich chemiluminescent magnetic microparticle immunoassay (CMIA) for quantitative detection of CrAg in sera. RESEARCH DESIGN AND METHODS: CMIA is based on magnetic beads modified with capture antibodies and biotinylated antibodies and Streptavidin-polyHRP, where biotinylated antibodies functioned as the recognition element and Streptavidin-polyHRP as the signal component. Assay parameters were first optimized, and then assay performances were evaluated. RESULTS: Under optimized conditions, the total runtime of the CMIA was 22 min. The assay had a wide linear range (2 -10,000 ng/mL) and high analytical sensitivity (0.24 ng/mL), together with acceptable reproducibility, accuracy, and stability. Besides, it exhibited no cross-reactivity with other pathogens. Importantly, the assay showed 92.91% (95% CI, 80.97-93.02%) overall qualitative agreement with a commercial ELISA kit in a retrospective cohort of 55 cases with confirmed cryptococcal infection, and 72 controls without evidence of invasive fungal disease (IFD). CONCLUSION: These results demonstrated that the present study paved a novel strategy for reliable quantitative detection of CrAg in sera.
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Antígenos Fúngicos , Criptococosis , Mediciones Luminiscentes , Humanos , Antígenos Fúngicos/sangre , Antígenos Fúngicos/inmunología , Mediciones Luminiscentes/métodos , Inmunoensayo/métodos , Criptococosis/diagnóstico , Criptococosis/sangre , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Femenino , Masculino , Persona de Mediana Edad , Adulto , Cryptococcus/inmunología , Estudios RetrospectivosRESUMEN
BACKGROUND: In July 2023, our hospital confirmed one case of lumbar spine infected complicated by Mycobacterium tuberculosis and Cryptococcus neoformans. The patient was admitted due to lower back pain for 1 year and a hard lump for 3 months. Symptoms and signs: Dressing can be seen fixed on the lower back, with severe bleeding. When the dressing is removed, a hard and protruding lump with a size of 6 cm x 8 cm, a sinus tract can be seen near the mass, with a slightly red wound and a sinus depth of about 3 cm. Light red fluid can be seen flowing out. There are no symptoms such as redness, swelling, or heat in the rest of the lower back, and the patient has no other underlying diseases or surgical history. METHODS: Lumbar magnetic resonance imaging and lumbar CT examination; Percutaneous puncture lumbar vertebral biopsy was performed, and the biopsy tissue was subjected to pathological examination, mNGS (metagenomic next-generation sequencing), and acid-fast staining; Extract pus from the lump for fungal culture and ink staining, and identify the fungi through MALDI-TOF MS. RESULTS: Bone destruction and bone marrow edema in the L5 vertebral body, compression of the spinal canal at the L5 vertebral body level; The pathological results of the biopsy tissue indicate granulomatous lesions. The acid-fast staining of the tissue is positive, and the mNGS of the tissue indicates infection with Mycobacterium tuberculosis. A single fungus was cultured from pus and identified by MALDI-TOF MS as Cryptococcus neoformans. Clinically, isoniazid 0.3 g ivgtt + rifampicin 0.45 g qd po + ethambutol 0.25 g qd po + pyrazinamide 0.75 g qd po + fluconazole 0.3 g qd po was administered for treatment. After 11 days, there was slight pain at the incision site, and the original symptoms were significantly relieved. The wound dressing was fixed in place, dry and without obvious exudation. Improved and discharged, followed up for 3 months with no recurrence of the lesion. CONCLUSIONS: mNGS is an effective identification technique that can be used to accurately diagnose suspected infection cases. MALDI-TOF MS has significant advantages over traditional detection methods in shortening detection time. This case achieved satisfactory treatment results for patients through a reasonable treatment plan, which is of great significance for exploring the diagnosis and treatment of similar disease infections.
Asunto(s)
Criptococosis , Cryptococcus neoformans , Vértebras Lumbares , Mycobacterium tuberculosis , Humanos , Cryptococcus neoformans/aislamiento & purificación , Vértebras Lumbares/microbiología , Mycobacterium tuberculosis/aislamiento & purificación , Criptococosis/diagnóstico , Criptococosis/microbiología , Criptococosis/tratamiento farmacológico , Masculino , Tuberculosis de la Columna Vertebral/diagnóstico , Tuberculosis de la Columna Vertebral/microbiología , Imagen por Resonancia Magnética , Antituberculosos/uso terapéutico , Antituberculosos/administración & dosificación , Persona de Mediana Edad , Tomografía Computarizada por Rayos XAsunto(s)
Cryptococcus , Meningitis Criptocócica , Humanos , Antifúngicos/uso terapéutico , Criptococosis/diagnóstico , Criptococosis/epidemiología , Criptococosis/inmunología , Criptococosis/terapia , Cryptococcus/inmunología , Cryptococcus/patogenicidad , Huésped Inmunocomprometido , Meningitis Criptocócica/diagnóstico , Meningitis Criptocócica/epidemiología , Meningitis Criptocócica/inmunología , Meningitis Criptocócica/terapia , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/inmunología , Interacciones Huésped-Patógeno/inmunología , Factores de Riesgo , Inmunosupresores/efectos adversos , Antígenos Fúngicos/sangre , Antígenos Fúngicos/líquido cefalorraquídeo , Hipertensión Intracraneal/etiología , Hipertensión Intracraneal/terapia , Síndrome Inflamatorio de Reconstitución Inmune/etiologíaRESUMEN
In vivo models provide advantages to study the progression of disease and to identify potential biomarkers to detect and monitor infections. For the human fungal pathogen Cryptococcus neoformans, murine intranasal models aim to recapitulate natural infection from inhalation of desiccated fungal cells from the environment and permit monitoring of disease over time. In this chapter, we describe the establishment of a murine model for cryptococcosis and the subsequent collection of organs, tissues, and fluids for sampling. These samples may support novel diagnostic strategies and opportunities to monitor dissemination of the fungal cells throughout the host and propose new treatment options to combat disease.
Asunto(s)
Criptococosis , Cryptococcus neoformans , Modelos Animales de Enfermedad , Animales , Cryptococcus neoformans/patogenicidad , Criptococosis/microbiología , Criptococosis/diagnóstico , Ratones , Manejo de Especímenes/métodos , HumanosRESUMEN
One of the standard assays for the fungal pathogen Cryptococcus neoformans is the glucuronoxylomannan (GXM) ELISA. This assay utilizes monoclonal antibodies targeted against the critical virulence factor, the polysaccharide (PS) capsule. GXM ELISA is one of the most used assays in the field used for diagnosis of cryptococcal infection, quantification of PS content, and determination of binding specificity for antibodies. Here we present three variations of the GXM ELISA used by our group-indirect, capture, and competition ELISAs. We have also provided some history, perspective, and notes on these methods, which we hope will help the reader choose, and implement, the best assay for their research.While it has long been referred to as the GXM ELISA, we also suggest a name update to better reflect our updated understanding of the polysaccharide antigens targeted by this assay. The Cryptococcal PS ELISA is a more accurate description of this set of methodologies and the antigens they measure. Finally, we discuss the limitations of this assay and put forth future plans for expanding the antigens assayed by ELISA.
Asunto(s)
Criptococosis , Cryptococcus neoformans , Ensayo de Inmunoadsorción Enzimática , Polisacáridos , Ensayo de Inmunoadsorción Enzimática/métodos , Cryptococcus neoformans/inmunología , Criptococosis/diagnóstico , Criptococosis/microbiología , Criptococosis/inmunología , Polisacáridos/análisis , Polisacáridos/inmunología , Humanos , Antígenos Fúngicos/inmunología , Antígenos Fúngicos/análisis , Polisacáridos Fúngicos/inmunología , Polisacáridos Fúngicos/análisis , Anticuerpos Monoclonales/inmunología , Anticuerpos Antifúngicos/inmunologíaRESUMEN
The importance of humoral immunity to fungal infections remains to be elucidated. In cryptococcosis, patients that fail to generate antibodies against antigens of the fungus Cryptococcus neoformans are more susceptible to the disease, demonstrating the importance of these molecules to the antifungal immune response. Historically, antibodies against C. neoformans have been applied in diagnosis, therapeutics, and as important research tools to elucidate fungal biology. Throughout the process of generating monoclonal antibodies (mAbs) from a single B-cell clone and targeting a single epitope, several immunization steps might be required for the detection of responsive antibodies to the antigen of interest in the serum. This complex mixture of antibodies comprises the polyclonal antibodies. To obtain mAbs, B-lymphocytes are harvested (from spleen or peripheral blood) and fused with tumor myeloma cells, to generate hybridomas that are individually cloned and specifically screened for mAb production. In this chapter, we describe all the necessary steps, from the immunization to polyclonal antibody harvesting, hybridoma generation, and mAb production and purification. Additionally, we discuss new cutting-edge approaches for generating interspecies mAbs, such as humanized mAbs, or for similar species in distinct host backgrounds.