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Ways to Quickly Identify Antibiotic Resistance in Bacterial Cells (Non-tradition

Proteins Genes Detection Cell Lysate PCR

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#1 Fluoresce

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Posted 25 January 2014 - 04:56 PM

Hello All,

Does anyone know of (a) good and sensetive molecular method(s) to identify the antibiotic resistence or sensetivity in bacterial cells directly from microlitter amount of sample without going throuhg 24-48 hours of work to grow the bacterial culture or plates in the begining?

I would be very thankful if you could help me on this. 

Thanks much in advance!

FL.



#2 pito

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Posted 26 January 2014 - 05:45 AM

no

 

You could run a PCR to look for the presence of genes encoding resistence, but thats pretty much it.


If you don't know it, then ask it! Better to ask and look foolish to some then not ask and stay stupid.

#3 Phil Geis

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Posted 26 January 2014 - 08:46 AM

As ;oito said, you could find the genetic signal but that wouldn't confirm expression and effective resistance.



#4 Fluoresce

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Posted 26 January 2014 - 09:47 AM

Thanks guys, that's what I was suspecting.

 

Now I am trying to figure out a few things:

 

How do Vitek2 and Phoenix phenotypically identify the bacterial suscep. in a short time?  Still need to read more on this but can anyone help here?

 

Any info on  16S rRNA is also being studied to identify the suscep/resist. of the bacterial strains? 



#5 pito

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Posted 27 January 2014 - 09:25 AM

16S rRNA is just to determine bacteria, not to figure out if they are suscep or res.

Dont forget that many of the resistance comes from plasmid DNA rather than genomic.

 

 

The Phoenix and Vitek2 just use standard tests (Bergeys) to identify bacteria, its actually just an "improved" way of the API strips.

For the suscep/resis : its just the same as doing it manually, but its just more automated and I guess a bit faster...

But it will still take time: you will still need to grow the bacteria etc....

 

 

Thanks guys, that's what I was suspecting.

 

Now I am trying to figure out a few things:

 

How do Vitek2 and Phoenix phenotypically identify the bacterial suscep. in a short time?  Still need to read more on this but can anyone help here?

 

Any info on  16S rRNA is also being studied to identify the suscep/resist. of the bacterial strains? 


If you don't know it, then ask it! Better to ask and look foolish to some then not ask and stay stupid.

#6 El Crazy Xabi

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Posted 27 January 2014 - 04:48 PM

For Vitek2 and Phoenix you still need to grow and isolate the bugs. It is only fast if you have them isolated and because you don't have to do the profiling manually nor prepare all the media. if you wanted to do something like a molecular method for that, you would need to integrate all the known potential elements that may indicate resistance, either DNA/RNA sequences or proteins (through specific antibodies), into a microarray... not easy stuff... and you will not cover sequences that are not discovered or divergent mutations, while in the present methods you only need the antibiotics, which in comparison are very limited in number



#7 Fluoresce

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Posted 27 January 2014 - 09:14 PM

Thank you both for the useful info.  Meanwhile I also read the documents provided by Biomerieux and BD.  Yes as Pito said they are still going through traditional methods and they need you to isoalte a single colony which is time consuming.  I need to cut this time down. 

So, El Crazy Xabi, do you know of good papers on the microarray format for bacterial ID/AST? 

Thanks for your help guys!



#8 pito

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Posted 28 January 2014 - 09:08 AM

You want micro-arrays to identify bacteria?

 

If so: they are out there, but most of them (if not all) are designed for specific bacteria: fish pathogens, plant pathogans, human pathogens......

SO it all depends on what you want ......

 

If you are speaking about the antibiotic resistance: it would not be that hard to design micro-arrays yourself based on resistance genes, but I doubt this really would work for what you want...

I mean: the gene can be present, but not active, plasmid vs genomic DNA, you would not know the levels it can tolerate....

Pretty much what EL Crazy Xabi also told: mutations in the genes, proteins to detect... it would be very complicated.


If you don't know it, then ask it! Better to ask and look foolish to some then not ask and stay stupid.

#9 Fluoresce

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Posted 28 January 2014 - 09:13 PM

Thanks Pito...I need to read a little more in this then...



#10 squint

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Posted 08 March 2014 - 07:25 PM

i did some research on this topic last year. You can get some information from these two studies and search more from related articles
Development of a DNA Microarray to Detect Antimicrobial Resistance Genes Identifiedin the National Center for Biotechnology Information Database(JG Frye,2010)
Analysis of Antimicrobial Resistance Genes Detected In Multidrug-Resistant Salmonella enterica Serovar Typhimurium Isolated from Food Animals(LSM Glenn, 2011)



#11 Fluoresce

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Posted 08 March 2014 - 07:33 PM

Thanks Squint...I appreciate your help.  I found the first paper.  Briefly went through it and it definitely will be helpful.  however, I don't have access to the second one.  Do you have access to this paper?  If you do, I would appreciate it if could please send the paper to my PM.  Thanks again for the info. 



#12 pito

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Posted 09 March 2014 - 12:08 AM

paper is in the attachement.

Its of course standard micro-array... not sure it is what you want unless you identify all the bacteria you "need" before you look for them.

 

Attached Files

  • Attached File  2.pdf   279.52KB   27 downloads

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#13 Fluoresce

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Posted 09 March 2014 - 08:06 AM

Thanks Pito. I need to put all pros and cons of various existing methods together for myself. Any good paper or experiences people have can help.





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