Diagnosis of Spontaneous Bacterial Peritonitis and an In Situ Hybridization Approach to Detect an “Unidentified” Pathogen

Figure 2

Schematic representation of the in situ hybridization (ISH) method used to assess the ascitic samples. Floating leukocytes in the ascitic fluid were collected via centrifugation and used for the ISH tests. DIG- (digoxigenin-) labeled probes were used for hybridization, and positive signals were detected with NBT (nitro-blue tetrazolium chloride) and BCIP (5-bromo-4-chloro-3′-indolyphosphate p-toluidine salt). Positive (purple brown) signals were observed in the leukocytes with intracellular bacterial DNA (arrows).