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Measuring microbiology in 30-40 seconds vs. ATP in 15 seconds

ATP Cleaning Validation Hygiene Testing Microbiology Bacteria

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#1 gskands

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Posted 30 January 2018 - 02:33 PM

Hi all

 

In short, I am part of a company where we have developed a small and affordable tech for real-time measurements of bacteria - we can currently measure total bacteria (non-specific, so similar to total viable count) in different aqueous solutions within 5 minutes. 

 

We are considering to develop a handheld device used with swabs to offer an alternative to ATP-measurements within hygiene and cleaning validation. The most important and risky development effort is lowering the measurement time without affecting the limit of detection for our current device. 

 

It is my impression that most ATP-sensors on the market today have a measuring time of approx. 15 seconds. How significant is this parameter when you evaluated different options for measuring ATP in hygiene and cleaning validation? 

 

I would argue the repeatability of our measurements is comparable to TVC measurements and much higher than ATP-testing. Furthermore, our preliminary testing with different cleaning agents and sanitizers do not provide false positives, which we have seen with ATP-devices. Our target is to reach a 30 seconds measurement time, but it is my concern that the market will dismiss the product if we can only get to e.g. 40 seconds as it will take the users too long to do testing (and holding something in your hands for 40 seconds is not ideal). 

 

I would love to hear your input on the matter. 

 

Best regards

Gustav



#2 t3hpr0phet

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Posted 30 January 2018 - 03:11 PM

While I don't see the duration of the test being a huge problem for someone who doesn't need to do many tests each session, a 30+ second test could result in a significant expenditure of time for someone with an extensive swabbing procedure.  Does the test read total cell count or total viable cell count? That could be a benefit that counteracts the extra time, but I'm not sure how much.



#3 adelk2202

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Posted 30 January 2018 - 03:46 PM

I am betting it would come down to cost. Would your testing cost less than an ATP system? I'm sure people would be willing to wait 20 extra seconds if they your system is cheaper. Also, as T3hpr0phet already mentioned if you only had a few swabs to do it wouldn't be as big of a deal. For myself we do 5 swabs in each of our RTE rooms and a little over one minute for our ATP swabs compared to almost 3.5 minutes for your system would be too big of a difference in time for the techs in the morning as they are doing these swabs right after they finish pre-op and before production begins set-up when time is of the essence. That is just my 2 cents though. I wish you luck because it sounds like a promising technology.



#4 gskands

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Posted 30 January 2018 - 04:12 PM

Very interesting input! These were exactly my concerns and I would love hear a few more opinions. To answer some of your questions, which might also help others:

 

Does the test read total cell count or total viable cell count? That could be a benefit that counteracts the extra time, but I'm not sure how much.

 

It detects all bacteria which are physically intact, i.e. viable. That being said there is a slight difference to plate counting. Incubating two identical samples at different temperatures with the standard TVC method will provide two different results - our technology counts all bacteria in the sample regardless of preferred growth conditions as we do not have an incubation or any other sample enrichment/preparation step, so our numbers are also higher than the typical plate count - it is a different base line to adjust to. If the bacteria membrane has somehow been perforated or otherwise destroyed, which is how most disinfectants kill bacteria, it will not be counted as a bacteria.

 

 

I am betting it would come down to cost. Would your testing cost less than an ATP system? I'm sure people would be willing to wait 20 extra seconds if they your system is cheaper. Also, as T3hpr0phet already mentioned if you only had a few swabs to do it wouldn't be as big of a deal. For myself we do 5 swabs in each of our RTE rooms and a little over one minute for our ATP swabs compared to almost 3.5 minutes for your system would be too big of a difference in time for the techs in the morning as they are doing these swabs right after they finish pre-op and before production begins set-up when time is of the essence.

 

Cost would be comparable to that of an ATP system. We do not intend to differentiate on cost as we believe we can add value through actually measuring bacteria instead of ATP. The argument is that bacteria is primarily what spoils food and poses a health risk, so it makes sense to measure the bacteria and use that as an indicator to gauge risk. I have a personal theory that ATP systems were only introduced because the alternative (measuring bacteria) was simply too slow and expensive - but that is a longer discussion for a different thread..



#5 Ryan M.

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Posted 31 January 2018 - 02:08 AM

If cost is comparable, I can see most companies pulling the trigger if the test is less than a minute.  I say a minute because if one is doing multiples swab sites don't they normally progress to the next sample site while the current sample is being tested?  At least one would think so...this way, it mitigates any lost time for the reader to read the sample.

 

If you make the cost comparable to ATP and 1 minute or less on the results time I would buy it right now.  Even if it was up to a few minutes on test time I would still buy it...why?  Because it is a MUCH MORE valuable tool, and I could always buy multiple readers to speed up the sampling and testing time.

 

Of course...anything you can do to improve the time and keep a good low level of detection is the greatest achievement.



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#6 w.weber

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Posted 31 January 2018 - 08:15 AM

Hello,

the system could have also another advantage. I am working in a company, where we can only do dry cleaning. So product rest may be still there. As ATP is showing up all ATP it has found, it is showing numbers, where we can't differ between high amount of bacteria or product rests detected. So if your system can differ between bacteria and e.g. plant cells, it could be a faster tool for us. 

Is it also planned to do different testing, not only tpc, perhaps also E.coli, Salmonellae ...? That would also give a more detailed and valuable view.

By the way, can you tell anything about the mechanism of detection? 



#7 Charles.C

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Posted 31 January 2018 - 12:45 PM

Hi gskands,

 

The name of the game is Validation. Some actual data or links to relevant publications might be useful.

 

I found yr criterion in Post4 rather disturbing from a safety POV -

 

The argument is that bacteria is primarily what spoils food and poses a health risk, so it makes sense to measure the bacteria and use that as an indicator to gauge risk.

 

(Perhaps you meant "and/or".)


Kind Regards,

 

Charles.C


#8 gskands

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Posted 01 February 2018 - 04:04 PM

If cost is comparable, I can see most companies pulling the trigger if the test is less than a minute.  I say a minute because if one is doing multiples swab sites don't they normally progress to the next sample site while the current sample is being tested?  At least one would think so...this way, it mitigates any lost time for the reader to read the sample.

 

If you make the cost comparable to ATP and 1 minute or less on the results time I would buy it right now.  Even if it was up to a few minutes on test time I would still buy it...why?  Because it is a MUCH MORE valuable tool, and I could always buy multiple readers to speed up the sampling and testing time.

 

Of course...anything you can do to improve the time and keep a good low level of detection is the greatest achievement.

 

I am glad to hear that you find the data output to be valuable, this also seem to be the general perception when we have talked to industry experts. However, our current system is a bench top device that can do the measurement in 5 minutes, which some beta customers also find very interesting, but so far systems have only been sold to research projects and "investigative" work (i.e. do a thorough analysis of where critical control points might be). The cost pr. sample for the current system is about 5-8x higher than ATP samples, so it is not priced to be part of routine operations. 

 

 

Hello,

the system could have also another advantage. I am working in a company, where we can only do dry cleaning. So product rest may be still there. As ATP is showing up all ATP it has found, it is showing numbers, where we can't differ between high amount of bacteria or product rests detected. So if your system can differ between bacteria and e.g. plant cells, it could be a faster tool for us. 

Is it also planned to do different testing, not only tpc, perhaps also E.coli, Salmonellae ...? That would also give a more detailed and valuable view.

By the way, can you tell anything about the mechanism of detection? 

 

To be honest it depends on how much plant residue you might have - if the surface is very dirty our system will clog. We do have tips and tricks to how you can run dirty samples though, simply pre-filter them through a 30-40 µm filter and you should be good to go. But the system definitely differentiates bacteria from plant cells (although we do not count the plant cells as they are too large to entire the measuring section of the device).  

 

Differentiation between different bacteria is in the works, but still at very early research stage. For now we only do total bacteria counts. Our method is non-destructive, so if you run a sample that has high counts then you can analyze it with a traditional e. coli/salmonella agar plate immediately after.

 

The detection mechanism is impedance flow cytometry. We have a small flow cell with electrodes whereon we measure a current. When a particle or bacteria crosses the electrodes we can immediately detect a unique change in current (at different frequencies), which can be used to identify the electrical properties of the particle. A bacteria is quite unique in its electrical composition when you consider cell membrane, cytoplasm, size, etc., and this is how we can differentiate bacteria from other particles in real-time. We need measurement time to establish statistical significance/evidence. Truthfully it is an advancement to the coulter-counter principle, where we have solved some pretty challenging issues within sample handling and measuring/differentiating bacteria in huge unknown matrices.

 

 

Hi gskands,

 

The name of the game is Validation. Some actual data or links to relevant publications might be useful.

 

I found yr criterion in Post4 rather disturbing from a safety POV -

 

(Perhaps you meant "and/or".)

 

Hi Charles.C,

 

Truthfully the solid white paper data we have for our current system is... not impressive. We have gathered quite a lot of data over the past 3 years from different experiments, but we have also done a million iterations on the product/tech, so we have postponed doing the huge third-party validation study until the product exits its prototype phase - mainly due to cost considerations. As previously mentioned the systems have only been sold to research projects and other detective work, so the sales effort often involves going meticulously through the lab data we have and do live demos (it is real-time after all). Of course we have simulated the validation studies in-house, and in other test environments, and everything looks great, but you will have to take my word for it for now. I hope this is OK since I am not trying to sell you or anyone else on this board the tech or a unit, I am just trying to get some input on what people would think about a system that measures total bacteria, is cost competitive to ATP, but slightly slower in measurement time!

 

For publications on the tech (impedance flow cytometry), you will quickly find that it has mostly been used to detect blood cells, which is easier than detecting bacteria because they are much larger. However, one article that shows an impedance system is used to detecting single bacteria is found here: http://pubs.rsc.org/...6G#!divAbstract. I think I have my name on one article in literature, but we only measured 0.5 µm, 1 µm and 2 µm particles with that system. We haven't published anything on the tech as a company because we do not want to give away any edge before we are able to fully support a product launch and gain a head start on competitors. There are currently no published articles on using impedance flow cytometry in a hygiene and cleaning validation setting, but you can expect that to change in 2018!

 

And you are entirely correct, I meant and/or. :)



#9 Ryan M.

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Posted 01 February 2018 - 04:31 PM

Some companies would still use it at 5-8 times higher cost over ATP.  Our company would.  Why you may ask?  We have production runs up to 72 hours making a large volume of products / SKU's.  Our finished QA results are 36 to 48 hours later.

 

If we can ensure we start with a clean system prior to startup it would be worth the additional cost / price, at least to us.

 

 

I am glad to hear that you find the data output to be valuable, this also seem to be the general perception when we have talked to industry experts. However, our current system is a bench top device that can do the measurement in 5 minutes, which some beta customers also find very interesting, but so far systems have only been sold to research projects and "investigative" work (i.e. do a thorough analysis of where critical control points might be). The cost pr. sample for the current system is about 5-8x higher than ATP samples, so it is not priced to be part of routine operations. 

 



#10 Charles.C

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Posted 03 February 2018 - 07:59 AM

Hi gskands,

 

Tks for yr reply.

 

It's an admirable objective but I am concerned that you may be (micro/chemically) comparing apples, oranges and pears. It's a question of correlation to existing defined "Standards".

 

There are also some additional items to navigate such as -

 

Relating satisfactory "hygiene" to a specific bacterial count is highly subjective, eg -

 

http://www.ifsqn.com...ces/#entry60958

 

And similarly for ATP (a parameter seemingly only occasionally correlatable to bacterial populations)

 

Nonetheless I look forward to seeing some data from yr Black Box !


Kind Regards,

 

Charles.C


#11 gskands

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Posted 05 February 2018 - 04:22 PM

Some companies would still use it at 5-8 times higher cost over ATP.  Our company would.  Why you may ask?  We have production runs up to 72 hours making a large volume of products / SKU's.  Our finished QA results are 36 to 48 hours later.

 

If we can ensure we start with a clean system prior to startup it would be worth the additional cost / price, at least to us.

 

Sounds interesting. Can I ask exactly what type of product you are producing? I would like to reach out to similar companies here in my own region and try to arrange pilot sales - that could generate some interesting data.

 

Hi gskands,

 

Tks for yr reply.

 

It's an admirable objective but I am concerned that you may be (micro/chemically) comparing apples, oranges and pears. It's a question of correlation to existing defined "Standards".

 

There are also some additional items to navigate such as -

 

Relating satisfactory "hygiene" to a specific bacterial count is highly subjective, eg -

 

http://www.ifsqn.com...ces/#entry60958

 

And similarly for ATP (a parameter seemingly only occasionally correlatable to bacterial populations)

 

Nonetheless I look forward to seeing some data from yr Black Box !

 

Hi Charles.C

 

With a fear of being branded as an outcast in this forum, then it is actually the "standards" we would like to challenge. The main objective for us to develop a solution that will conclusively provide more value and thus safety to our clients and customers by improving the existing hygiene and cleaning validation regardless of what hygiene standards are defined in different countries today - if we only consider the standards (and methods used in the standards) then I think it will be difficult or impossible to ever improve. Measuring total bacteria, and being well aware that it is different from CFU, is what we hypothesize to be the best solution for performing hygiene and cleaning validation, simply because it says how much bacteria/microbiology is actually on the surface and therefore is great at risk assessment, and it can provide the fast result that is necessary to take action. This hypothesis needs to be backed by data, and this is what we are in the process of generating both through validation studies (soon) and with early adopters (now). I will be happy to share along the way - but please tell me if I step out of line in regards to promoting what we do. :)

 

I am fully aware we have a long road ahead of us, and I hope to have a positive back and forth with you and this forum along the way as your input and concerns are very likely to represent the food industry as a whole. We need to be able to address these, otherwise we do not have market relevance. Therefore, I appreciate your input and please keep the comments flowing in future threads!

 

BR

Gustav



#12 Charles.C

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Posted 05 February 2018 - 07:59 PM

 

Hi Charles.C

 

With a fear of being branded as an outcast in this forum, then it is actually the "standards" we would like to challenge. The main objective for us to develop a solution that will conclusively provide more value and thus safety to our clients and customers by improving the existing hygiene and cleaning validation regardless of what hygiene standards are defined in different countries today - if we only consider the standards (and methods used in the standards) then I think it will be difficult or impossible to ever improve. Measuring total bacteria, and being well aware that it is different from CFU, is what we hypothesize to be the best solution for performing hygiene and cleaning validation, simply because it says how much bacteria/microbiology is actually on the surface and therefore is great at risk assessment, and it can provide the fast result that is necessary to take action. This hypothesis needs to be backed by data, and this is what we are in the process of generating both through validation studies (soon) and with early adopters (now). I will be happy to share along the way - but please tell me if I step out of line in regards to promoting what we do. :)

 

I am fully aware we have a long road ahead of us, and I hope to have a positive back and forth with you and this forum along the way as your input and concerns are very likely to represent the food industry as a whole. We need to be able to address these, otherwise we do not have market relevance. Therefore, I appreciate your input and please keep the comments flowing in future threads!

 

BR

Gustav

 

Hi Gustav,

 

As per yr OP, I get the impression you are mainly proposing yr methodology as an alternative to ATP rather than as a competitor to current, lengthy, culture based counts. Clearly time is the critical factor for many current users of ATP methodology.

 

From yr OP -

I would argue the repeatability of our measurements is (a) comparable to TVC measurements and (b) much higher than ATP-testing

 

For (a,b) swabbing error would presumably be comparable. And typically (relatively) high? (obviously it may relate to the specific surface type/condition)

 

TVC afaik has typically a Low level of accuracy as reflected in, existing, decision criteria. IMEX this is an intrinsic limitation.

 

ATP measurement error no idea (not a user) ? i deduce you consider it to be << than TVC. Any particular reason ?

 

I would have thought yr (static) counting method should have a much higher accuracy than TVC ? So what is the limitation ?


Kind Regards,

 

Charles.C


#13 gskands

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Posted 06 February 2018 - 11:11 AM

Hi Gustav,

 

As per yr OP, I get the impression you are mainly proposing yr methodology as an alternative to ATP rather than as a competitor to current, lengthy, culture based counts. Clearly time is the critical factor for many current users of ATP methodology.

 

From yr OP -

 

For (a,b) swabbing error would presumably be comparable. And typically (relatively) high? (obviously it may relate to the specific surface type/condition) 

It is true that the operator is one of the most important factors when you consider swabbing errors. To my knowledge it is very common that it is the same operator who performs the procedure at different production sites as it reduces the variance introduced by the swabbing method.

 

TVC afaik has typically a Low level of accuracy as reflected in, existing, decision criteria. IMEX this is an intrinsic limitation.

This is not my impression from 19 customer interviews. Of course this is a limited data set, but it is my impression that most customers prefer to use TVC for accurate measurements, but use ATP as they need to the speed to react. I know of some companies that only monitor with TVC, because they did not trust the ATP results when they tested the method. If this is due to lack of education or training I do not know. Using TVC only allows them to do trend analysis, and they are not able to react immediately on hygiene issues.

 

ATP measurement error no idea (not a user) ? i deduce you consider it to be << than TVC. Any particular reason ?

This is where we quickly end up comparing apples and oranges. The ATP instrument is developed to measure and detect organic material, which may indicate hygiene issues, but correlating the presence of organic material to actual risk is a stretch, especially when you compare correlating TVC measurements to actual risk. This is exemplified in Ryan M.'s case where they can not use ATP, because there will always be some plant residue after cleaning, but that does not necessarily mean there is a risk in the production. I would argue that we do hygiene and cleaning validation to determine and reduce risk in the production, and while ATP is a good tool, there are better parameters (such as bacteria) to use for risk assessment. The problem today is that it just simply takes too long to measure bacteria. That being said, you can not do a 1:1 correlation between bacteria count and risk either, but I would argue the correlation is stronger than for ATP. 

 

I would have thought yr (static) counting method should have a much higher accuracy than TVC ? So what is the limitation ?

I would argue that it has, but we are up against a 100 year old method and a science (microbiology) that heavily relies on routine. We are working a lot on how to tell "the story", in the sense that we often get questions like "why don't we only care about the specific bacteria that can grow?" and "Are there really that much bacteria on surfaces? We don't see that using the methods we have for a long time." The answer to question 1 is that whatever result you get with a TVC will change if you change the temperature or the non-specific agar media, so you are not measuring which bacteria can grow, you are only measuring which bacteria can grow under the provided conditions. Your product might be subjected to an entirely different environment throughout the supply chain, so there is a risk some of the bacteria you did not see with a TVC will start growing and pose a risk. The answer to question 2 can be validated by looking at literature and reminding people of the great plate count anomaly, which is text book stuff, but easily forgotten.

 

Everything about "the story" has to be backed by data and reference cases to really make an impact on the industry. Right now it is only early technology adopters who engage with us, as they recognize our train of thought.

 

Sorry, I got carried away in our discussion. It is true that for now we propose our tech to be a replacement for ATP, but it is a long-term strategy to also replace TVC-measurements in hygiene and cleaning validation. I think it is best to focus our discussion on the ATP-part, because this is what we believe we can improve on short term. However, my comments to your questions are found in red above.


Edited by Charles.C, 09 February 2018 - 02:20 AM.
font re-formatted


#14 gskands

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Posted 06 February 2018 - 11:12 AM

It appears I made a formatting error so some of my answers aren't highlighted. Is it possible to edit a post?



#15 GMO

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Posted 06 February 2018 - 01:04 PM

I know some people swear by it but I'm not much of a "believer" in ATP.  I've found very variable results and that they don't correlate very well with lab testing.  If your method does correlate well then I think it has a place but as other have said it will come down to cost.  Some people use ATP routinely so if it's comparable cost to ATP they may switch to your method given enough evidence of efficacy.  Personally I think ATP is far too expensive for the benefit but your method may be something I use in response to an incident or for training if it correlates well with traditional lab methods.  

A 40 second test time wouldn't worry me at all.  Anything under 1 minute is sufficiently fast for me but if a person is doing a lot of swabs to verify a clean they will want the fastest result they can get.



#16 Ryan M.

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Posted 06 February 2018 - 03:26 PM

Sounds interesting. Can I ask exactly what type of product you are producing? I would like to reach out to similar companies here in my own region and try to arrange pilot sales - that could generate some interesting data.

 

 

Hi Charles.C

 

With a fear of being branded as an outcast in this forum, then it is actually the "standards" we would like to challenge. The main objective for us to develop a solution that will conclusively provide more value and thus safety to our clients and customers by improving the existing hygiene and cleaning validation regardless of what hygiene standards are defined in different countries today - if we only consider the standards (and methods used in the standards) then I think it will be difficult or impossible to ever improve. Measuring total bacteria, and being well aware that it is different from CFU, is what we hypothesize to be the best solution for performing hygiene and cleaning validation, simply because it says how much bacteria/microbiology is actually on the surface and therefore is great at risk assessment, and it can provide the fast result that is necessary to take action. This hypothesis needs to be backed by data, and this is what we are in the process of generating both through validation studies (soon) and with early adopters (now). I will be happy to share along the way - but please tell me if I step out of line in regards to promoting what we do. :)

 

I am fully aware we have a long road ahead of us, and I hope to have a positive back and forth with you and this forum along the way as your input and concerns are very likely to represent the food industry as a whole. We need to be able to address these, otherwise we do not have market relevance. Therefore, I appreciate your input and please keep the comments flowing in future threads!

 

BR

Gustav

 

We produce extended shelf-life fluid products; dairy and non-dairy.  We see typical shelf-life of 75 days to 110 days, all paperboard.  There are about 20 facilities which do this currently in the USA.  Another process, very similar to us, is the aseptic bottling / paperboard processes.  The only difference is our packaging lines are not considered aseptic, but the rest of the process is aseptic.  Both process types have extended runs over 24 hours up to 72 hours depending on the product.  We also need to verify surfaces are absolutely clean prior to startup because 72 hours is A LOT OF PRODUCT.



#17 Charles.C

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Posted 09 February 2018 - 03:22 AM

Hi Gustav,

 

I am just trying to get some input on what people would think about a system that measures total bacteria, is cost competitive to ATP, but slightly slower in measurement time!

 

It is obvious that the working situation for some of previous posters would make them immediately interested in yr device/its response time if  accurate, practical and of reasonable cost. Others would perhaps need to be convinced of the comparative benefits for their work scenarios.

 

I still recall the first time i used Atomic Absorption instruments for certain difficult product analyses. Operatively simple, rapid, validatably accurate and selective. Pure magic !. But the sensitivity for certain elements was insufficient until later refinements became available.

 

Initially ATP was believed to be routinely correlated to TVC data. Unfortunately this belief was later shown to require validation for the specific situation. ATP's  practical sensitivity to different units / products / environments is evident from supplier instrument manuals. Nonetheless where applicable, the technique has obviously delivered highly satisfactory results to many people.

 

So IMHO it all comes back to Validation / Data.

 

Good Luck !

 

PS - sorry for delay, didn't see yr last Post


Kind Regards,

 

Charles.C






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