Potential clinical benefit of the in situ hybridization method for the diagnosis of sepsis

Development of a new in situ hybridization method for the detection of global bacterial DNA to provide early evidence of a bacterial infection in spontaneous bacterial peritonitis

“…In addition, leukocytes which ingested fungus were also prepared and used as negative control samples. The procedures used for the ISH test were performed as previously described [9][10][11], and positive signals were detected in the cellular cytoplasm as purple-brown stains.…”

“…In addition, a representative smear sample of a fungus (C. albicans: NBRC1594) was also prepared as a negative control sample. The slides were fixed for 20 min in Carnoy's solution, and ISH was performed according to the methods described previously [9][10][11].…”

“…In order to identify the causal bacteria in sepsis, we previously reported a novel strategy using an in situ hybridization (ISH) method to detect the genomic DNA of bacteria phagocytized in neutrophils and macrophages [9][10][11]. Our previous studies established the utility of the ISH method by demonstrating evidence supporting the presence of a bacterial infection.…”

A New Colorimetric Peptide Nucleic Acid-Based Assay for the Specific Detection of Bacteria

“…It was observed that shorter periods allowed an easy discrimination of a positive signal from the background noise, but the highest S/N was obtained for 45 min of hybridization (Figure 3). This hybridization period is actually very short comparing to hybridization times used for other ISH procedures using EUB338 or other DNA probes, which usually take between 6 and 18 h [2,28].…”

“…However, rapid detection is still difficult to achieve, especially using conventional techniques. It has been demonstrated that pathogenic microorganisms could be detectable with in situ hybridization (ISH) and that this method is useful for the accurate and rapid diagnosis of infectious agents, especially in blood samples [1,2]. A number of labeled DNA or RNA probes and hybridization protocols are available to detect target nucleic acids of pathogens, especially in clinical specimens, but these techniques are usually laborious [3].…”