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PCR methods of microbe testing in food industry, Is it worth the time and money?

#PCR #pathogen #testing #lab

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Jat

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Posted 22 October 2021 - 06:05 PM

We are finalizing to setup a small in site lab to run our pathogen testing with PCR system. Except of the cost analysis that I did , I'm still thinking if it is worth of it or not. 

1- first concern is about enrichment process for Salmonella, Listeria and Campylobacter. It needs some preparations for media before enrichment which I really hate.

2-  Second is about cross contamination, as we might not be able to provide HVAC system  and biosecurity cabinet at this time for our tiny lab. I do not know how much risk there might be with PCR system.(before lysis step)

 

I'll be appreciated if any expert in this area can provide me some tips before final decision to purchase these pricy equipment.

Thanks,



Charles.C

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Posted 22 October 2021 - 07:24 PM

We are finalizing to setup a small in site lab to run our pathogen testing with PCR system. Except of the cost analysis that I did , I'm still thinking if it is worth of it or not. 

1- first concern is about enrichment process for Salmonella, Listeria and Campylobacter. It needs some preparations for media before enrichment which I really hate.

2-  Second is about cross contamination, as we might not be able to provide HVAC system  and biosecurity cabinet at this time for our tiny lab. I do not know how much risk there might be with PCR system.(before lysis step)

 

I'll be appreciated if any expert in this area can provide me some tips before final decision to purchase these pricy equipment.

Thanks,

Hi Jat,

 

No direct experience with PCR but you must have an enviable budget.

And, I would anticipate, some rather knowledgeable support/resources.

Lucky You ! :thumbup:


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Kind Regards,

 

Charles.C


kingstudruler1

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Posted 22 October 2021 - 08:07 PM

My wife has the 3M MDS.    https://www.3m.com/3...ection-systems/

Not going to lie, its pretty ccol.  It seems like they have interesting financing options.    

 

your two concerns are legitimate for anyone looking to do inhouse pathogen testing.   you will always have to enrich.  if you don't think you can properly control the cross contamination risk, its probably not worth it.    


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Jat

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Posted 22 October 2021 - 08:29 PM

Hi Jat,

 

No direct experience with PCR but you must have an enviable budget.

And, I would anticipate, some rather knowledgeable support/resources.

Lucky You !

 Hi Charles, you can't imagine how much time and effort I put to convince GM for this cool technology.  :smile:

Also, initial cost was lower than what I expected at first. besides running costs easily can outweigh Ext lab costs.

I guess there would be more devices available in the future to facilitate in site food testing mostly with short product shelf life like ours. 


Edited by Jat, 22 October 2021 - 08:30 PM.


Jat

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Posted 22 October 2021 - 08:40 PM

My wife has the 3M MDS.    https://www.3m.com/3...ection-systems/

Not going to lie, its pretty ccol.  It seems like they have interesting financing options.    

 

your two concerns are legitimate for anyone looking to do inhouse pathogen testing.   you will always have to enrich.  if you don't think you can properly control the cross contamination risk, its probably not worth it.    

Hi Kingstudruler1,

Thanks for your input,

Basically all of these PCR systems are the same, just different brands. I was digging all of them from Hygiena, 3M, Bio-rad ,etc for months and review their features.

Unfortunately, our budget will be exhausted for the equipment this year. I do not know how much risk you might face with not having a biosecurity hood/ HVAC system. 

I was thinking because samples are transferred from the bag by multichannel pipettors to test tubes, which are enclosed there would be a minimal risk of contamination.

Just my assumption  :smile:



kingstudruler1

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Posted 22 October 2021 - 09:00 PM

Hi Kingstudruler1,

Thanks for your input,

Basically all of these PCR systems are the same, just different brands. I was digging all of them from Hygiena, 3M, Bio-rad ,etc for months and review their features.

Unfortunately, our budget will be exhausted for the equipment this year. I do not know how much risk you might face with not having a biosecurity hood/ HVAC system. 

I was thinking because samples are transferred from the bag by multichannel pipettors to test tubes, which are enclosed there would be a minimal risk of contamination.

Just my assumption  :smile:

 

I think the  risk is lower as well.  it would be ideal to have a hood.   the far greater risk is contamination from people and items moving  in and out of the lab to other areas of the facility.  



juanolea1

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Posted 22 October 2021 - 10:37 PM

I think that you are underestimating the risks. Cultivating Food Pathogens can be risky because of the risks associated with cross contamination. It involves many steps, from pre-enrichment to enrichment, testing and sterilizing the resulting incubated samples. I believe that the greatest risk comes from personnel mobility. Contamination can happen through spillage on the floor, on equipment and contaminated space. Sooner or later employees can carry a cell or two in or on their shoes.

When you incubate food samples you tend to incubate and concentrate the target organisms at around 10^3 and 10^4 CFU/ml (sometimes higher) to get a decent amplification (Malorny, et al., 2004). I know I am citing an old article and that current detection levels may be lower, but current incubation protocols lead to as many organisms. Lab spills happen all the time and technicians can carry the unwanted pathogens into many common areas of the facility.

Even if you decide to test in-house you need to become accredited on a continuous basis for your results to be officially recognized. The accreditation process takes time, money and effort to put in place.

When you add all related costs associated with testing and the potential risk of contamination it may not seem like a good idea, but everyone knows their own capabilities. If you decide to test in-house, ensure that the testing facility is not in the same location as the plant and that you follow proper decontamination steps for people visiting the testing building and the facility.

I hope it helps!

 

Juan

 

Reference:

 

Malorny, B., Paccassoni, E., Fach, P., Bunge, C., Martin, A., & Helmuth, R. (2004). Diagnostic real-time PCR for detection of Salmonella in food. Applied and environmental microbiology, 70(12), 7046–7052. https://doi.org/10.1....7046-7052.2004


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Jat

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Posted 22 October 2021 - 10:54 PM

I think that you are underestimating the risks. Cultivating Food Pathogens can be risky because of the risks associated with cross contamination. It involves many steps, from pre-enrichment to enrichment, testing and sterilizing the resulting incubated samples. I believe that the greatest risk comes from personnel mobility. Contamination can happen through spillage on the floor, on equipment and contaminated space. Sooner or later employees can carry a cell or two in or on their shoes.

When you incubate food samples you tend to incubate and concentrate the target organisms at around 10^3 and 10^4 CFU/ml (sometimes higher) to get a decent amplification (Malorny, et al., 2004). I know I am citing an old article and that current detection levels may be lower, but current incubation protocols lead to as many organisms. Lab spills happen all the time and technicians can carry the unwanted pathogens into many common areas of the facility.

Even if you decide to test in-house you need to become accredited on a continuous basis for your results to be officially recognized. The accreditation process takes time, money and effort to put in place.

When you add all related costs associated with testing and the potential risk of contamination it may not seem like a good idea, but everyone knows their own capabilities. If you decide to test in-house, ensure that the testing facility is not in the same location as the plant and that you follow proper decontamination steps for people visiting the testing building and the facility.

I hope it helps!

 

Juan

 

Reference:

 

Malorny, B., Paccassoni, E., Fach, P., Bunge, C., Martin, A., & Helmuth, R. (2004). Diagnostic real-time PCR for detection of Salmonella in food. Applied and environmental microbiology, 70(12), 7046–7052. https://doi.org/10.1....7046-7052.2004

Thanks Juan for your great points.

Hopefully the lab is in a separate building and away from process and products and even operating staff. 

We are a poultry processing plant and our products normally carry salmonella, campylobacter and E.coli  And you're right, with enrichment process we actually multiply it by thousands time. Our priority is to have an annual CCP validation through product testing. It's important to set a baseline first and try to reduce indicators in our products (Qualitative and Quantitative). 

We have got so many contradictions with Ext Lab results from previous years data. So, it made us to go this way.






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