ABSTRACT
AIMS: We established a real-time PCR assay for the detection and strain identification of Candida species and demonstrated the ability to differentiate between Candida albicans the most common species, and also Candida parapsilosis, Candida glabrata, Candida tropicalis and Candida dubliniensis by LightCycler PCR and melting curve analysis. METHODS AND RESULTS: The DNA isolation from cultures and serum was established using the QIAmp Tissue Kit. The sensitivity of the assay was ≥ 2 genome equivalents/assay. It was possible to differentiate all investigated Candida species by melting curve analysis, and no cross-reaction to human DNA or Aspergillus species could be observed. CONCLUSIONS: The established real-time PCR assay is a useful tool for the rapid identification of Candida species and a base technology for more complex PCR assays. SIGNIFICANCE AND IMPACT OF THE STUDY: We carried out initial steps in validation of a PCR assay for the detection and differentiation of medically relevant Candida species. The PCR was improved by generating PCR standards, additional generation of melting curves for species identification and the possibility to investigate different specimens simultaneously.
Subject(s)
Candida/classification , Polymerase Chain Reaction/methods , Candida/genetics , Candida/isolation & purification , Candida albicans/genetics , Candida albicans/isolation & purification , Candida glabrata/genetics , Candida glabrata/isolation & purification , Candida tropicalis/genetics , Candida tropicalis/isolation & purification , DNA, Fungal/isolation & purification , HumansABSTRACT
Skin involvement due to a systemic infection with Aspergillus species in the course of allogeneic hematopoietic cell transplantation is extremely rare. We report the clinical course of two patients with hematologic malignancies who underwent allogeneic hematopoietic cell transplantation (HCT) and developed disseminated skin involvement as the first clinical symptom of a proven systemic Aspergillus infection.
Subject(s)
Aspergillosis/pathology , Hematopoietic Stem Cell Transplantation/adverse effects , Skin Diseases/etiology , Adult , Aspergillosis/etiology , Aspergillosis/immunology , Fatal Outcome , Hematologic Neoplasms/complications , Hematologic Neoplasms/therapy , Humans , Immunocompromised Host/physiology , Male , Middle Aged , Necrosis , Skin Diseases/microbiology , Skin Diseases/pathology , Transplantation, Homologous/adverse effectsABSTRACT
A 37-year-old male patient with a diffuse pleomorphic B-cell-lymphoma, which has been diagnosed two month earlier with the primary site at the pterygopalatine fossa on both sides with infiltration of the clivus and cavernous sinus was referred to our hospital for continuation of the third course of CHOP chemotherapy. At admission he reported about a recent history of painful swallowing and intermittent substernal chest pain. Alleviation of the pain on swallowing and the chest pain was apparently only possible by drinking 10 to 15 l of cold coca cola throughout the day and night, a regimen that resulted in polyuria. Physical examination revealed extensive thrush stomatitis and soor esophagitis. Despite successful treatment with fluconazole, polydipsia continued unabated. The classic osmotic test of dehydration and exogenous vasopressin revealed hypothalamic diabetes insipidus (DI). Basal hormones and stimulated endocrine function tests of the adenohypophysis were found to be normal. MRI-scan revealed lymphoma infiltration of the neurohypophysis. After the third course of CHOP chemotherapy the patient surprisingly recovered completely from his excessive thirst. The present report shows that clinical disorders such as thrush stomatitis can mask diabetes insipidus caused by an early relapsing lymphoma.
Subject(s)
Diabetes Insipidus/diagnosis , Lymphoma, B-Cell/complications , Pituitary Neoplasms/complications , Adult , Antifungal Agents/therapeutic use , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Candidiasis, Oral/drug therapy , Candidiasis, Oral/etiology , Cyclophosphamide/therapeutic use , Diabetes Insipidus/etiology , Diabetes Insipidus/therapy , Diagnosis, Differential , Doxorubicin/therapeutic use , Esophagitis/etiology , Fluconazole/therapeutic use , Humans , Lymphoma, B-Cell/diagnosis , Lymphoma, B-Cell/drug therapy , Magnetic Resonance Imaging , Male , Neoplasm Recurrence, Local , Pituitary Gland, Posterior , Pituitary Neoplasms/diagnosis , Polyuria , Prednisone/therapeutic use , Stomatitis/etiology , Thirst , Vincristine/therapeutic useABSTRACT
Acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) are thought to arise from malignant hematopoietic progenitor cells representing early and undifferentiated stem cell clones. In CML there is evidence for a progenitor cell subset free of leukemic clones, depending on the course of the disease. Additionally, it has been suggested that in AML, the early stem cell compartment (CD34+/90+) does not harbor the malignant clone. We analyzed white blood cells from leukemia patients for the presence of aberrant cells in stem cell subfractions. Sixteen patients with CML, six patients with AML, two patients with acute lymphatic leukemia (ALL) and one with chronic myelomonocytic leukemia (CMMOL), all with known cytogenetic abnormalities, were evaluated according to their CD90 (Thy-1)-positive or -negative phenotype. Subsets were sorted on to slides and further characterized by FISH and/or standard cytogenetic testing. The bcr-abl translocation or gross chromosomal abnormalities could be detected in equally high amounts of 92.2% and 89.2% in both stem cell subsets. We conclude, that in progressed AML and CML cells characterized by specific genetic aberrations implicated in the malignant state can be found in the CD34+/CD90+ and CD90- population, thus making CD90 an inappropriate marker to distinguish benign from malignant cells in these leukemias.
Subject(s)
Antigens, CD34/analysis , Chromosome Aberrations/genetics , Hematopoietic Stem Cells/immunology , Hematopoietic Stem Cells/pathology , Leukemia, Myeloid/genetics , Thy-1 Antigens/analysis , Acute Disease , Flow Cytometry , Humans , In Situ Hybridization, Fluorescence , Karyotyping , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/blood , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/pathology , Leukemia, Myeloid/blood , Leukemia, Myeloid/pathologyABSTRACT
We report on two elderly female patients who experienced musical hallucinations. Both were suffering from deafness in addition to non-specific organic brain changes. In both patients the musical hallucinations were successfully treated with carbamazepine.
