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"... Scientists at the Ulsan National Institute of Science & Technology (UNIST) in South Korea have unveiled their novel diagnostic technique known as fluorescence in situ hybridization (FISH), using artificial polymers – peptide nucleic acid (PNA) – that act as probes to bind to different genetic sequences within bacteria. When the two probe molecules bind to the target, fluorescent signals are emitted, which essentially reveal the fingerprint of different pathogens. ...
And FISH can produce these results in less than three hours. Conventional diagnostic tools – blood tests and polymerase chain reaction (PCR) analyses can take days to weeks. ..."
"... We developed a novel set of peptide nucleic acid (PNA)-based FISH probes with optimal target specificity by analyzing the variations in 16S ribosomal RNA sequence across all bacterial species." ...
This innovative FISH method employs two PNA molecules simultaneously. By analyzing the genomic sequences of 20,000 bacterial species, the research team designed PNA sequences that specifically target the ribosomal RNA of particular species. PNA exhibits a higher sensitivity to sequence mismatches compared to conventional DNA-based probes and demonstrates superior penetration through bacterial cell walls. Furthermore, the requirement for both PNA molecules to bind to their target site before generating a signal significantly reduces the likelihood of crosstalk, thereby enhancing accuracy in situations involving multiple overlapping bacterial strains. ..."
From the abstract:
"Fast and accurate identification of pathogenic microbes in patient samples is crucial for the timely treatment of acute infectious diseases such as sepsis.
The fluorescence in situ hybridization (FISH) technique allows the rapid detection and identification of microbes based on their variation in genomic sequence without time-consuming culturing or sequencing. However, the recent explosion of microbial genomic data has made it challenging to design an appropriate set of probes for microbial mixtures.
We developed a novel set of peptide nucleic acid (PNA)-based FISH probes with optimal target specificity by analyzing the variations in 16S ribosomal RNA sequence across all bacterial species. Owing to their superior penetration into bacteria and higher mismatch sensitivity, the PNA probes distinguished seven bacterial species commonly observed in bacteremia with 96–99.9% accuracy using our optimized FISH procedure.
Detection based on Förster resonance energy transfer (FRET) between pairs of adjacent binding PNA probes eliminated crosstalk between species. Rapid sequential species identification was implemented, using chemically cleavable fluorophores, without compromising detection accuracy. Owing to their outstanding accuracy and enhanced speed, this set of techniques shows great potential for clinical use."
Fast and accurate multi-bacterial identification using cleavable and FRET-based peptide nucleic acid probes (no public access)
Figure 1. Schematic representation illustrating the key findings of the study.
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