Howie Rose wrote:
> Has anybody had experience of using RNA probes
>for the detection of pathogenic bacteria without an amplification
>system? As there are many copies of RNA floating around a cell, I
>would have thought that a detection system with the appropriate
>sensitivity highlight these without having to amplify as is necessary
>with DNA.
Dear Howie
We have experience with fluorescently labelled oligonucleotide
riboprobes (both DNA and RNA constructs) targeted to either 16S rRNA
genomic DNA (as a taxonomic tool) or to expressed 16S rRNA (as a
diagnostic/viability indicator). These probes have been used
successfully in the detection of Legionella spp in biofilm systems. As
there are some 10>4 - 10>5 ribosomes/cell (ie ~40-50% cell mass in
rapidly growing cells) then an 'amplification' effect is elicited in
growing cells (obviously reduced in VNC status). Please contact me if
you need more information
Kevin Bown
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Kevin Bown,
Centre for Applied Microbiology & Research,
Porton,
Wilts, SP4 OJG. UK.
Tel: +44(0)1980 612666
Fax: +44(0)1980 612731
e-mail: kevin.bown at camr.org.uk
pager: 01426 157893
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