Two rapid dual color fluorescence in situ hybridization (FISH) assays were evaluated for detecting M. tuberculosis and related pathogens in cultures. The MN Genus-MTBC FISH assay uses an orange fluorescent probe specific for the Mycobacterium tuberculosis complex (MTBC) and a green fluorescent probe specific for the Mycobacterium and Nocardia genera (MN Genus) to detect and distinguish MTBC from other Mycobacteria and Nocardia. A complementary MTBC-MAC FISH assay uses green and orange fluorescent probes specific for the MTBC and M. avium complex (MAC) respectively to identify and differentiate the two species complexes. The assays are performed on acid-fast staining bacteria from liquid or solid cultures in less than two hours. Forty-three of 44 reference mycobacterial isolates were correctly identified by the MN Genus-specific probe as Mycobacterium species, with six of these correctly identified as MTBC with the MTBC-specific probe and 14 correctly as MAC by the MAC-specific probe. Of the 25 reference isolates of clinically relevant pathogens of other genera tested, only four isolates representing two species of Corynebacterium gave a positive signal with the MN Genus probe. None of these 25 isolates were detected by the MTBC and MAC specific probes. A total of 248 cultures of clinical mycobacterial isolates originating in India, Peru and the USA were also tested by FISH assays. DNA sequence of a part of the 23S ribosomal RNA gene amplified by PCR was obtained from 243 of the 248 clinical isolates. All 243 were confirmed by DNA sequencing as Mycobacterium species, with 157 and 50 of these identified as belonging to the MTBC and the MAC, respectively. The accuracy of the MN Genus-, MTBC-and MAC -specific probes in identifying these 243 cultures in relation to their DNA sequence-based identification was 100%. All ten isolates of Nocardia, (three reference strains and seven clinical isolates) tested were detected by the MN Genus-specific probe but not the MTBC- or MAC-specific probes. The limit of detection for M. tuberculosis was determined to be 5.1x104 cfu per ml and for M. avium 1.5x104 cfu per ml in liquid cultures with the respective MTBC- and MAC-specific probes in both the MN Genus-MTBC and MTBC-MAC FISH assays. The only specialized equipment needed for the FISH assays is a standard light microscope fitted with a LED light source and appropriate filters. The two FISH assays meet an important diagnostic need in peripheral laboratories of resource-limited tuberculosis-endemic countries.
Oral mucositis (OM)-related outcomes constituting a meaningful clinical advance in bone marrow transplant patients were considered by an interdisciplinary panel. Meaningful outcomes are essential in product development for OM, a condition without effective prevention or treatment. The most important outcomes to measure, the feasibility of measuring these in a clinical trial, and clinically meaningful differences in these outcomes were determined by the panel. Most important are reduction in oral pain and use of opioid analgesics, improvement in oral intake and quality of life, and reduction of hospitalization duration. Reduction in the severity of OM measured by an objective evaluation of oral mucosa could provide insight regarding the biologic activity of an intervention. Further data are required to define the precise relationship between reduction in visible OM and improvement in outcome. Minimally, clinical trials for OM should assess oral pain, opioid use, oral intake, and include objective assessment of OM.
dRapidly growing mycobacteria are rarely found in central nervous system infections. We describe a case of polymicrobial infection in a brain abscess including two rapidly growing Mycobacterium species, M. immunogenum and M. llatzerense. The Mycobacterium isolates were distinguishable by molecular methods, and whole-genome sequencing showed <60% pairwise nucleotide identity. CASE REPORTA 40-year-old woman with a past medical history notable for migraines and frequent childhood sinopulmonary infections presented with 7 days of unremitting right frontal headache and 2 days of night sweats, chills, vomiting, left-sided vision loss, and right leg numbness. Unlike her prior migraines, this headache was more intense and not relieved by rest in a dark room; it was also associated with visual impairments and sensation abnormalities. Throughout childhood, the patient had suffered from recurrent upper and lower respiratory tract infections, for which she had received frequent antibiotics, chest percussion, and sinus irrigation using a neti pot with unfiltered, unsterilized tap water. As an adult, the frequency of her respiratory infections decreased to approximately annually, with her last one being 4 months prior to admission. The patient was born and raised in rural Pennsylvania and had regularly consumed unpasteurized milk and drunk well water. In the preceding months, the patient had swum in a river in northern California and traveled to France, where she had consumed various soft cheeses.Upon presentation, the patient was moderately distressed but alert and oriented to person, place, and time. She had a temperature of 37°C, a heart rate of 51 beats per minute, a blood pressure of 122/66 mm Hg, a respiratory rate of 12 breaths per minute, and an oxygen saturation of 100% on room air. Her physical exam was remarkable for a left lower homonymous quandrantanopia and reduced sensation to light touch on her right lower leg. She had symmetric facial musculature, a clear oropharynx, no paranasal or frontal sinus tenderness, and no neck stiffness. Her lungs were clear to auscultation bilaterally, cardiac exam demonstrated sinus rhythm with a soft systolic murmur, musculoskeletal strength and reflexes were within normal limits, and there were no skin lesions. A complete blood count was notable for a white blood cell count of 11.3 ϫ 10 9 /liter (90% neutrophils), hemoglobin at 11.2 g/dl, and a platelet count of 287 ϫ 10 9 /liter. The C-reactive protein level was 7.4 mg/liter, and an erythrocyte sedimentation rate was 30 mm/h. T1-weighted magnetic resonance imaging (MRI) of the brain demonstrated a 3.7-cm by 2.8-cm by 3.0-cm rim-enhancing lesion in the right parietal/occipital lobe, with restricted diffusion upon diffusion-weighted MRI (Fig. 1). The patient underwent emergent craniotomy and drainage of pressurized purulent fluid contained within the cavity.Gram staining demonstrated Gram-positive cocci and Grampositive rods, and the patient was started on empirical antibiotic therapy with vancomycin, metronidazole, and ceftriaxo...
We report the draft genome sequence of Mycobacterium obuense strain UC1 from a patient sputum sample. This is the first draft genome sequence of Mycobacterium obuense, a rapidly growing scotochromogenic mycobacterium.
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