Bacterial ennum. by PCR

Zophonias Oddur Jonsson zjons at rhi.hi.is
Tue Dec 14 08:42:46 EST 1993


In <dowe.22.2D0D3D42 at ace.acadiau.ca> dowe at ace.acadiau.ca (MARK DOWE) writes:

>Hi all,

>     I'm working on a procedure to enumerate a species of bacteria in soil 
>using PCR, and I was wondering if anyone had any experience/advice that 
>they would be willing to share?
>     Specifically, I'm working with Listeria monocytogenes and primers for 
>the listeriolysn gene.  I was hoping to use the procedure in studies of a 
>possible viable but nonculturable state in the organism. 

PCR (and LCR) has been used to detect and discriminate betwen Listeria strains
In all the work that I am aware of the 16S rDNA is used but I guess the
listeriolysin could be used as well.  

Mabe you already have these references but it can´t do any harm to post them:

 1.	Golsteyn-Thomas, Elizabeth J., Robin K. King, et al., Sensitive and Specific
Detection of Listeria monocytogenes in Milk and Ground Beef with the Polymerase
Chain Reaction. Applied and Environmental Microbiology, 1991. 57:9(September):
p. 2576-2580.  

2.	Deneer, Harry G. and Irene Boychuk, Species-Specific Detection of listeria
monocytogenes By DNA Amplification. Applied and Environmental Microbiology,
1991. 57(2): p. 606-609.  

3.	Bubert, Andreas, Stephan Kohler, et al., The Homologous and Hererologous
Regions within the ap Gene Allow Genus- and Species-Specific Identification of
Listeria spp. by Polymerase Chain Reaction. Applied and Environmental
Microbiology, 1992. 58(8): p. 2625-2632. 

4.	Czajka, Joh, Nada Bsat, et al., Differentiation of Listeria monocytogenes
and Listeria innocua by 16S rRNA Genes and Intraspecies Discrimination of
Listeria monocytogenes Strains by Random Amplified Polymorphic DNA
Polymorphisms. Applies and Environmental Microbiology, 1993. 59(January): p.
304-308.  

---->  Here is another Idea, use the Ligase Chain Reaction.  If you want more
info on that there are some more references I can give you.  

5.	Wiedmann, Martin, John Czajka, et al., Discrimination of Listeria
monocytogenes from Other Listeria Species by Ligase Chain Reaction. Applied and
Environmental Microbiology, 1992. 58:11(November): p. 3443-3447.  


I hope this helps
 
Zophonias O. Jonsson
University of Iceland




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