case_images: PMC10928390_01
case_images: (a) The Computerized Axial Tomography (CAT) scan exhibited a consolidation in the right maxillary sinus and notable thickening of the mucosa in both the left maxillary and sphenoid sinuses. (b) Microscopic examination of Bipolaris spp. microculture (lactophenol blue stain, 400x) and microculture on sabouraud agar.
case_images: acmi-6-743.v3-g001.jpg
case_images: PMC10928390_01_acmi-6-743.v3-g001.jpg
case_images: F1
case_images: A computerized axial tomography (CAT) scan disclosed complete consolidation in the right maxillary sinus and partial involvement of the ethmoidal sinuses (Fig. 1a). A sample was collected to investigate a suspected opportunistic infection, revealing the presence of septate hyphae (Fig. 1b).
case_images: (a) Quellung's reaction: isolate 25 041, classified as NT-Spn, exhibited no reactivity in any of the anti-serum tests. In contrast, isolate 26 378, identified as S. pneumoniae serotype 3 (positive control), yielded a positive reaction with the specific anti-serum. (b) Gel electrophoresis of S. pneumoniae-specific PCR products targeting the 295 bp lytA gene. Lane 1: negative control; lane 2: isolate 25041 NT-Spn; lane 3: isolate 26 378 S. pneumoniae serotype 3. And 160 bp cpsA gene Lane 1: negative control; lane 2: isolate 25041 NT-Spn; lane 3: isolate 26 378 S. pneumoniae serotype 3. (c) Negative stain (phosphotungstic acid). On the left the Isolate 25041 NT-Spn demonstrates loss of the capsule compared with the image on the right, isolate 26 378 S. pneumoniae serotype 3, with the polysaccharide capsule surrounding the cell of the pneumococcal isolate.
case_images: acmi-6-743.v3-g002.jpg
case_images: PMC10928390_01_acmi-6-743.v3-g002.jpg
case_images: F2
case_images: In our case, the isolate did not react with any of the pools described (Fig. 2a), therefore in order to realize a full analysis of the isolate, a molecular approach was applied. Firstly, a conventional PCR targeting the lytA gene was conducted, yielding a positive result (Fig. 2a). The result was negative (Fig. 2b). Following the introduction of a counterstain (methylene blue), pneumococcal cells become dark blue, enclosed by a well-defined halo representing the outer boundary of the capsule (Fig. 2a). Negative staining with phosphotungstic acid provided high contrast images, enabling the visualization of fine structural details of the samples at the nanometer scale (Fig. 2c).