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#1 Pizza&Sandwich

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Posted 29 March 2013 - 05:47 PM

I have been using ATP testing for a while to determine if the cleaning has been sufficient. Recently I ordered the Neogen allergen tests for Egg. I swabbed the same areas with ATP & Allergen test: ATP: 3.6 (>3.0 = fail), Allergen = negative. Can someone shed some light on what might be going on here? How can we be removing the egg allergen, but not getting the surface clean?
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#2 abhagat

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Posted 29 March 2013 - 06:45 PM

That's because ATP tests are as high as 10 times more sensitive at detecting proteins as compared to the Neogen allergen tests.


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#3 SPL

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Posted 29 March 2013 - 06:57 PM

What type of material are you swabbing from? These ATP swabstest measures for free ATP that maycome from any organic material or bacterial present. ATP may be from microbialsthat were not destroyed during your clean procedure, mold and yeast. RLUs/ATPlevels do not directly correlate to microbial counts and present of allergen(as you saw in your test)


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#4 Pizza&Sandwich

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Posted 29 March 2013 - 07:40 PM

Stainless steel and plastic conveyor belts.
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#5 KTD

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Posted 01 April 2013 - 03:36 PM

Lacey -

If you swabbed the 'same' areas, is there a potential for cross-reactivity or chemical interference between the tests?
It is possible that you removed the egg proteins but only partially destroyed a biofilm that then reacted with the ATP test only.
Are you using the lateral flow or ELISA-style allergen testing? I have recently had detection issues with Neogen's lateral flow test kits...


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#6 Pizza&Sandwich

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Posted 01 April 2013 - 03:39 PM

Lacey -

If you swabbed the 'same' areas, is there a potential for cross-reactivity or chemical interference between the tests?
It is possible that you removed the egg proteins but only partially destroyed a biofilm that then reacted with the ATP test only.
Are you using the lateral flow or ELISA-style allergen testing? I have recently had detection issues with Neogen's lateral flow test kits...



Lateral flow. Tell me more about your detection issues.


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#7 Tony-C

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Posted 01 April 2013 - 07:15 PM

I have been using ATP testing for a while to determine if the cleaning has been sufficient. Recently I ordered the Neogen allergen tests for Egg. I swabbed the same areas with ATP & Allergen test: ATP: 3.6 (>3.0 = fail), Allergen = negative. Can someone shed some light on what might be going on here? How can we be removing the egg allergen, but not getting the surface clean?


Hi Lacey,

ATP swabbing and Allergen tests are not the same. The ATP swabs measure free ATP from any source (Not protein - there are other swabs that measure protein/sugar) & the Egg Allergen Test is specific and solely measures Egg Protein.

Your results are feasible, load and content of residue on your conveyors could be a factor, it would be useful to know the exact tests you are using.

Regards,

Tony
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#8 KTD

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Posted 01 April 2013 - 07:41 PM

Lacey -

I have seen
- negative soy tests when the ingredient contains either hydrolyzed or other severely processed soy products
- lateral flow devices have a smaller detection range than the ELISA-style...depending on your food matrix and allergen concentration, the latteral flow test kit can be overloaded when verifying that the test kit works for your products prior to cleaning validation

On the other side of the coin
- some manufacturers are labeling products that have been considered/labeled 'allergen-free' (certain food oils & lecithins and products with extremely low allergen concentrations) as containing an allergen, but it cannot be detected
- lateral flow tests are less sensitive than the ELISA-style, so for validating cleaning activities, the ELISA-style may work better for you


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#9 SPL

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Posted 01 April 2013 - 08:53 PM

Lacey -

I have seen
- negative soy tests when the ingredient contains either hydrolyzed or other severely processed soy products
- lateral flow devices have a smaller detection range than the ELISA-style...depending on your food matrix and allergen concentration, the latteral flow test kit can be overloaded when verifying that the test kit works for your products prior to cleaning validation

On the other side of the coin
- some manufacturers are labeling products that have been considered/labeled 'allergen-free' (certain food oils & lecithins and products with extremely low allergen concentrations) as containing an allergen, but it cannot be detected
- lateral flow tests are less sensitive than the ELISA-style, so for validating cleaning activities, the ELISA-style may work better for you



Here a presentation poster by neogen on the hook effect.

But here a break down of how each test looks for.

Elisa and Lateral Flow Assay - target is specific antigens of allergen. They are ensitive to 5 - 10 ppm depend on test and manufacture

Protein Swab - multiple allergen proteins eg. milk, egg, soy and etc sensitive to micrograms

ATP Swab - looks for ATP. ATP is the basic engery unit of cells and may carry over on proteins.
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#10 mgourley

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Posted 01 April 2013 - 10:08 PM

This shows why I am not a fan of ATP testing.
Unless you have done significant baseline ATP testing to set up Pass/Fail criteria, you are basically just looking at a random number.
Is the surface really not clean of you show a >3 on your ATP swab? If the result is 100, probably not, but then again, what exactly are you looking for?

Allergens are another matter entirely. If you are looking for a negative allergen test, but there are (and probably will be) ATP present, which is more important? Lack of allergens on the test or is the surface "clean"?

Marshall


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#11 Oldairyman

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Posted 02 April 2013 - 07:34 PM

I tend to agree with Marshall, If you have residuals of Fats or sanitizers on the surface it will affect the results of ATP testing. Is it really clean ?
Jeff
:dunno:

This shows why I am not a fan of ATP testing.
Unless you have done significant baseline ATP testing to set up Pass/Fail criteria, you are basically just looking at a random number.
Is the surface really not clean of you show a >3 on your ATP swab? If the result is 100, probably not, but then again, what exactly are you looking for?

Allergens are another matter entirely. If you are looking for a negative allergen test, but there are (and probably will be) ATP present, which is more important? Lack of allergens on the test or is the surface "clean"?

Marshall


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#12 SPL

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Posted 02 April 2013 - 07:49 PM

Forget to attach this file

Attached Files


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#13 john.kukoly

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Posted 03 April 2013 - 04:00 PM

Very educating thread, many thanks to the obviously well informed contributors. A few additions to the ongoing debate around post sanitation validation and verification of allergen cleaning:

- keep in mind the allergen concentration in the formula - if the allergenic protein represents 0.1% of the product matrix, one would expect similar results in ATP vs protein specific. Your result may be showing that there is an opportunity for better cleaning that protein specific testing does not illustrate.

- variance in test reagent solubility of the soil complex - each reagent and physical sample collection tool will pick up different compounds at different rates. The same goes for the actual cleaning process; not all soils are equally removed by all methods, leaving a residue different in make up than the original soil load

- microbiological loading post cleaning - depending on the time and conditions, the bio load may increase prior to testing, giving a noticeable ATP signature while the allergen protein signature remains low to undetectable.

Once you take into account the protein test methodology mentioned earlier, there is nothing wrong ith utilizing both test methods during validation, looking to the combined result to show sufficient removal of all relevant soil loads.


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#14 Charles.C

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Posted 03 April 2013 - 05:53 PM

Dear All,

As intimated in earlier posts, as far as ATP is concerned with respect to verification of surface cleanliness (unrelated to allergen considerations), we are perhaps avoiding the primary issue, -

Define a clean surface.

If one considers a plate count measurement as the gold standard, the published ("satisfactory") criteria can vary anywhere between (typically) 2.5 and 100 cfu / cm2. Some micro. textbooks simply define it as specific to one's own product / system when "satisfactorily" cleaned.

Based on published studies, it seems unarguable that there is a significant number of authors who consider that ATP is simply not a suitable technique for verifying cleanliness due various factors, most of which have already been mentioned on this forum. Although specific criteria for non-applicability seem lacking.
It is also maybe significant that after 20 yrs or so, AFAIK, there is no commercial ATP device which is AOAC approved (??)

As an example i found the 2009 document quoted below rather depressing although no detailed data is given.

ATP-monitoring Systems
Using an ATP-monitoring system for ICM in situ comparative studies is both logical and economical. ATP is a rapid and accurate method for establishing baseline values of biocontaminant cleanliness. However, a word of caution is in order. By last count, there are approximately five manufacturers of ATP systems that supply the American market; I have four of them in my arsenal of field instruments and use each for a different and specific application in my work.

Each of my units differs from the others in almost every aspect, such as the swab systems and chemistries, the light-sensing devices, the software programs and readouts, the key-pad operation and their levels of sensitivity and methods of calibration. In fact, their diversity of design helps me in my profession. One of my units is multifunctional and can be used for measuring temperature, pH and conductivity. This is ideal when bulk sampling presents a safety hazard. Another unit can be used with multipurpose swab types. These provide me with a wide field of study, including water and allergen surrogate assays. Two of my units are very easy to operate, extremely economical and ideal for teaching purposes. Of these two units, one has a unique swabbing system that I can use on a filter medium with an air-sampling device (but more on this in a future column). In addition, three of my units are programmable, and collective data can be downloaded to my laptop computer. Two sampling systems are not overly temperature-sensitive and therefore nicely adaptable to field work. The list goes on.

The only downside to this diversity is that there exists no current standard matrix whereby applying a constant or “fudge factor” will yield comparable relative light unit (RLU) results for all systems. The only comparisons that I have seen so far in the industrial literature basically relate to their relative sensitivities and environmental interferences, shelf lives of the swab systems, ease of use and various bells and whistles unique to each manufacturer. Having said all this, there is yet another bit of information of which you should be aware when working with this technology. Nowhere in the scientific literature are any standard numbers given for cleanliness. Those that have been reported are unique to that study, with all surfaces and situations being different. Each area and surface is different from every other and should be evaluated by establishing separate baseline values and then comparing the results of cleaning activities against those values. As a general rule in enumerating microbial assays, only orders-of-magnitude differences should be considered significant. What all this means is that if you start with a given manufacturer’s unit, continue with that same unit or one from the same manufacturer. If you adapt another manufacturer’s ATP-monitoring system, you need to reestablish baseline values for cleanliness in each experiment.

Further Lessons Learned
Unlike microbiological assays where surface sampling detects some but not all living bacteria, ATP-monitoring systems detect everything. Since ATP is present in all living cells, these systems will detect anything of either plant or animal origin, including all microbes, living or dead. Therefore, establishing controls before sampling is sometimes more important than the experiment itself and must be done to rule out inaccuracies in data collection.

For instance, water used in cleaning may pass through a resin, charcoal or calcite bed or some other filtering system and may contain large numbers of heterotrophic bacteria or their equivalent cell content. Likewise, surface-water sources may have elevated ATP levels from algae, bacteria and fungi that normally live in the source water impoundment. Defining these background levels is essential to the interpretation of the results. I learned this the hard way.

http://www.foodsafetymagazine.com/magazine-archive1/aprilmay-2009/the-me-in-icm-using-atp-to-evaluate-sanitation/

And similarly this publication -
Attached File  Comparison atp and micro. swabbing ca 2003.pdf   137.72KB   28 downloads

Maybe there are some equally pro-publications around also. :smile:

Rgds / Charles.C
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Kind Regards,

 

Charles.C


#15 mgourley

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Posted 03 April 2013 - 06:46 PM

I read this article a few years ago and validates my "anti-ATP" stance.
Basically the user has to determine what "clean" is, and then do baseline tests with ATP swabs and equipment to verify that a surface is "clean".
Seems to me that since organoleptic methods were probably used to determine what "clean" is, why not just continue to use those methods to validate your cleaning procedure?

Yes, I know everyone has different products/facilities/equipment and the prospect of an "instant" pass/fail test is alluring.
Bottom line, IMO, is that ATP testing really does not tell you anything about cleanliness and as a result is worthless in the real world.

Sort of like metal detection as a CCP :whistle:

Marshall


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#16 Tony-C

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Posted 03 April 2013 - 07:35 PM

Bottom line, IMO, is that ATP testing really does not tell you anything about cleanliness and as a result is worthless in the real world.

Marshall


:yikes: :potplant:

Putting this politely, you are in a small minority, ATP testing is widely recognised as a useful tool in assessing cleaning performance.

BRC Global Standard for Food Safety:
Acceptable level of cleaning may be defined by ...............ATP bioluminescence techniques (also referred to in CIP process verification)

The AIB International Consolidated Standards for Inspection Prerequisite and Food Safety Programs:
Post-cleaning evaluation techniques, which could include....................Adenosine triphosphate (ATP) testing
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#17 mgourley

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Posted 03 April 2013 - 08:13 PM

Yes, Tony.

But as you posted, "may" and "could" is not exactly a ringing endorsement.

Since there is no recognized standard as to what ATP testing actually measures, in relation to "cleanliness", I don't find it useful.

I'm certainly not saying people should not use it. I just don't see the value add in it. And yes, I've spent time and money in the past using it.

In my particular case, (bread, buns) I can tell when a mixer is clean just by looking at it.
Other peoples mileage may of course, vary.

Marshall


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#18 Pizza&Sandwich

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Posted 03 April 2013 - 08:54 PM

Yes, Tony.

But as you posted, "may" and "could" is not exactly a ringing endorsement.

Since there is no recognized standard as to what ATP testing actually measures, in relation to "cleanliness", I don't find it useful.

I'm certainly not saying people should not use it. I just don't see the value add in it. And yes, I've spent time and money in the past using it.

In my particular case, (bread, buns) I can tell when a mixer is clean just by looking at it.
Other peoples mileage may of course, vary.

Marshall


Since you're in the bread/bun industry, what type of conveyor belts do you use for your dough? How do you clean them?
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