ATP Products

Why do I get a different RLU result when I re-measure the same test device again in the luminometer?

You will always get a different reading when trying to re-read the same test device more than once. The meter is reading a light given off by the reagent. This light peaks and then begins to die off. Always measure your test device in the time recommended in the product instructions. Any result you receive after the first reading may be misleading because that light will continue to die out until eventually no light is emitted.

How does temperature affect ATP test result?

Ambient temperature of 20–22°C (70–72°F) is the temperature that provides optimal performance. The only time ambient temperatures can affect results is if reagents are at a low temperature. This can happen if testing is done immediately after taking the tests out of the refrigerator. If testing in cold environments the instrument will self-adjust to external temperature but the luciferase reagents needs to be at  20–22°C (70–72°F) to function best.

If reagents are cold, then the reaction will be slower and the RLU result for a given amount of ATP will be lower.

Temperature does not affect light measurement of the instrument provided that the instrument has equilibrated to the environmental conditions. The instrument will sense an environmental temperature change of 5°C and it will automatically initiate a 15 second recalibration sequence.

The user can also initiate a manual recalibration at any time by depressing and holding the OK button down for 3 seconds.

The system can be used in cold environments (e.g. 5°C) if the instrument is equilibrated for 10 – 15 minutes before use and that the reagent swab devices are kept warm e.g. by storage in an internal pocket close to body heat.

Will direct sunlight affect test results?

Yes. Tests activated and measured in direct sunlight will distort results, increasing the RLU output. If a test must be collected outside, the swab device should be activated and measured out of the direct sunlight.

What are Hygiena ATP swabs pre-moistened with? Is it safe?

Hygiena ATP test swabs are pre-moistened with a mild extraction solution that aids in sample collection and ATP extraction. The solutions is non-toxic and is not a growth media, so it is completely safe. Please refer to the Safety Data Sheet for the product for further information about safety.

Can I compare RLU measurement to CFU plate counts?

ATP systems with standard ATP test swabs detect the presence of ATP and cannot distinguish microbial ATP from other organic ATP left behind after cleaning. RLU results could potentially be microbial cells, organic ATP from product residue, or both. This makes comparing RLU to CFU difficult. Studies have shown that there is anywhere between 60-90% correlation between RLU and CFU readings depending on the environment in which the system is used.

The presence of ATP on the surface indicates that it has not been adequately cleaned. The primary purpose of cleaning is to remove product residue for product contact surfaces. Effective cleaning simultaneously removes the material capable of supporting microbial survival and growth, as well as many of the microbes themselves. The technology behind ATP systems has been changing quite rapidly over the past few years and now there are ATP systems that can run tests specifically for microorganisms.

For more information on these multi-functional ATP systems see the MicroSnap platform from Hygiena. 

Can I have different limits for different locations?

Yes. Pass/Fail limits are 100% customizable to fit your facility and program needs. To adjust limits for specific locations, simply edit the Unit Set-Up data in the SureTrend software. 

How do I set my Pass and Fail limits for my ATP program?

All systems come preset with generic limits that can usually be applied to most processing facilities. These thresholds have been established over the years by companies doing ATP testing and supported by research papers on the science. These thresholds are typically a good starting point for companies new to ATP testing.

Click here for a video and step by step guide for setting up and determining RLU limits!

What is the limit of detection of Hygiena ATP tests?

The limit of detection depends on the test device and luminometer used:

  • UltraSnap & SystemSURE Plus : 1 femtomole of ATP
  • SuperSnap & SystemSURE Plus : <0.20 femtomoles of ATP
  • SuperSnap & EnSURE : <0.10 femtomoles of ATP

Does a 0 (zero) RLU result indicate sterility?

Sterile is a term relating to a true microbiology test i.e., the absence of living bacteria. An ATP test detects all sources of ATP but cannot differentiate ATP from food or bacteria, thus a zero RLU result cannot be equated with sterility.

Are there any studies or white papers on ATP testing?

You can find third party research, editorials, reports, and other documents on product pages of this website. Many manufacturers offer seemingly scientific research supporting their own ATP systems. It is important for customers to understand that only objective studies conducted by independent laboratories should be accepted as scientific evidence of product performance. Silliker Laboratories, the leading international food testing and safety laboratory, conducted an independent study comparing five commercial ATP systems. This study found that Hygiena’s products offer superior linearity, sensitivity, repeatability, and accuracy. Request a copy here!

Do all foods contain ATP?

Because ATP is present in all organic matter, all foods contain ATP at varying levels. This is why testing for ATP presence is the preferred way to measure the cleanliness of surfaces and water samples. Highly processes foods such as oil will have very little to no ATP.

How many food and beverage processing companies are doing ATP monitoring?

Nearly all food and beverage companies that process food with any water activity or can sustain organisms do ATP testing. Companies with highly processed materials like oil or companies dealing with products with no water activity like dry grains, sometimes will not do ATP testing. ATP testing is used around the globe and is a recognized tool by auditors for validating cleaning processes and complies with government and HACCP regulations.

What is ATP Monitoring?

ATP monitoring is a rapid testing method used by many companies to quickly assess the cleanliness of surfaces or liquid samples. Adenosine Triphosphate (ATP) is present in all organic material and is the universal unit of energy used in all living cells. ATP is produced and/or broken down in metabolic processes in all living systems. Processes such as photosynthesis in plants, muscle contraction in humans, respiration in fungi, and fermentation in yeast are all driven by ATP. Therefore, most foods and microbial cells will contain some level of naturally occurring ATP.

Hygiena luminometers (in conjunction with ATP swabs) use bioluminescence to detect residual ATP as an indicator of surface cleanliness. The presence of ATP on a surface indicates improper cleaning and the presence of contamination, including food residue, allergens and/or bacteria. This implies a potential for the surface to harbor and support bacterial growth.

ATP monitoring is used in food and beverage facilities to confirm that ATP presence is eliminated or minimized by effective sanitation procedures. ATP monitoring prevents cross-contamination, ensures product integrity, potentially improves product shelf life, protects brand reputation, and complies with regulatory requirements.

How does bioluminescence work?

Bioluminecsence is the result of a biochemical reaction and is the science behind Hygiena ATP detection tests. The reaction includes the following elements:

  • Luciferase enzyme- naturally occurring in fireflies, or synthetically manufactured
  • Adenosine triphosphate (ATP) – the energy molecule of all living organisms
  • Oxygen- a catalyst
  • Luicferin- a molecule that undergoes a chemical charge when affixed by an enzyme

ATP + Luciferin + Luciferase + O2 == == Light output

The reaction occurs in two steps: The substrate combines with ATP and oxygen, which is controlled by the enzyme. The chemical energy in step 1 excites a specific molecule (the combination of Luciferin and Luciferase). The result is decay which is manifested as photon emission, or light production. The light is simply a by-product of the chemical reaction and does not depend on light.

How do I contact Hygiena to purchase an ATP solution?

Hygiena products are available worldwide. We have a direct sales staff in the USAUK and parts of Europe, and Asia. Hygiena also has a network of dedicated distributors and agents across the globe, covering over 80 countries. 

Why are Hygiena’s products so much more cost effective?

Hygiena strives to create easier-to-use, more cost-effective products for its clients. By partnering with leading manufacturing companies and using state-of-the-art automated manufacturing processes in house, Hygiena is able to create superior cost-effective products. A perfect example of this is our ATP bioluminescence product line. It incorporates patented and environmentally-friendly Snap-Valve™ technology, which allows us to create an easy-to-use sample test device that uses less plastic than our competition and has superior performance. Combining this with our unique liquid-stable reagents allows us to have a complete all-in-one sample testing device that has better performance and costs a lot less. Compare systems side by side or see performance data here. 

How long are ATP tests good for?

Hygiena ATP tests have a 15 month shelf life at refrigerated temperatures (2-8°C) and 4 week shelf life at room temperature (21-25°C). All swab labels are printed with expiration month and year for your convenience.

How do I properly dispose of Hygiena ATP test devices?

Hygiena ATP test devices are made of 100% recyclable, non-toxic plastic and may be discarded with plastic recyclables.(Recycling Code 7:Other).

What is the proper swabbing technique?

You should swab an area that is about 4 by 4 square inches (10 x 10 cm) – or in the case of a hard-to-clean area, as much of the surface as possible. Do not let the swab come into contact with anything other than the test area to avoid contamination. Apply pressure to the swab to pick up surface residue and penetrate any biofilm that may be present. After collecting the sample, place the swab back in swab tube. Once the device has been activated, it should be read a soon as possible.

Correct Incorrect

Sufficient pressure to create flex in
the swab shaft

Do not touch the swab shaft

Rotate to collect sample on all sides
of swab tip

Only lightly swabbing the surface

Touching the swab shaft with finger

Only collecting sample on one side 
of swab tip

How often should critical and regular test sites be swabbed?

Critical (high-risk) test sites should be swabbed on a daily basis, after each cleaning or after terminal cleaning. If a failure is measured, immediate corrective action should be taken with re-cleaning and re-testing until a passing result can be measured. Regular (lower-risk) control points may not need to be tested as frequently.

What areas should be tested?

ATP testing can be used on any surface where cleanliness needs to be confirmed. This may include patient rooms, surgical tools and equipment, staff keyboards and work stations, operating rooms, waiting rooms, high-touch areas, etc. The CDC provides a list of recommended test locations for monitoring environmental cleaning. For a list of test areas, refer to the System Implementation Guide or contact a Hygiena representative for assistance.

What is the difference between cleaning, disinfection, and sterilization?

Cleaning is the removal of organic matter and the reduction of risk from material which is a potential contaminant, or material which could support the survival and growth of microbes. 

Disinfection  is the reduction of microbiological hazards to a minimal level but not necessarily complete inactivation of all microbiological hazards. 

Sterilization is the complete destruction and inactivation of all microbial hazard. 

Will sanitizers or soap interfere with my swab counts?

Several sanitizers commonly used in hospitals have been tested at normal working strength and found to have no significant effect on either SnapShot or UltraSnap performance. Only acid-based sanitizer, if used at higher than manufacturer-recommended concentrations have been found to have an effect on performance. If you are unsure about the chemical nature of your sanitizer, consult your sanitizer manufacturer or Hygiena technical support.

Does a surface have to be dry before I can swab?

Surfaces do not have to be dry to perform a swab test. However, for consistent readings, surfaces should be swabbed in the same conditions (always wet or always dry). Hygiena swabs come pre-moistened for maximum sample recovery on dry surfaces.

When should I do ATP testing?

ATP testing should ideally be done after cleaning, but before sanitizer or hydrogen peroxide vapor is applied. Because sanitizers are less effective when product residues are on the surface, it is best to eliminate all ATP before the sanitizer step. In some facilities, testing after cleaning is not possible. In these scenarios testing after sanitizing is acceptable.

What is biofilm?

A biofilm is formed when microorganisms find a receptive environment where they are exposed to food and moisture. The microorganisms work together as a population and secrete a sticky polymer to form a solid matrix attached to a surface. Once a biofilm is established, it is very difficult to eliminate because the microbes are reinforced and protected by the matrix, making them very resistant to sanitizers. The threat of a biofilm can be eliminated with proper ATP hygiene monitoring, allowing early detection and removal of organic residue – thus eliminating the food source for possible biofilm-forming microbes. In addition, UltraSnap ATP swabs have a unique detergent on the swab tip that cuts through biofilm and exposes the underlying cells. If a biofilm has already developed, there will be more ATP on a surface, which will result in a higher RLU and it can be detected by the SystemSURE Plus and eliminated with vigorous cleaning.

Can I compare RLU counts to CFU plate counts?

The SystemSURE Plus luminometer detects ATP presence and cannot distinguish microbial cells from other organic materials left behind after cleaning. Because RLU count could potentially be a representation of microbial cells as well as other organic materials, RLU’s cannot be equated to a CFU plate count. However studies have shown about an 80% correlation between ATP and microbial cells. The presence of ATP on the surface indicates that it has not been adequately cleaned. The primary purpose of cleaning is to remove germs and bacteria from environmental surfaces. Effective cleaning simultaneously removes the material capable of supporting microbial survival and growth, as well as many of the microbes themselves.

What will I see on the luminometer screen after taking an ATP test?

Below is an image of the luminometer screen:

Can I have different limits for different locations?

Yes. Pass/Fail limits are 100% customizable to fit your facility and program needs. To adjust limits for specific locations, simply edit the location data in SureTrend software.

How do I establish my Pass/Fail RLU limits?

Hygiena provides industry-standardized recommendations for broad risk categories such as near patient areas, public areas, etc. You may also set your own Pass/Fail RLU limits by collecting samples from test locations. For a full guide on establishing Pass/Fail RLU limits, refer to the System Implementation Guide that comes with your system, or contact a Hygiena representative for assistance.

Can I use ATP to verify the effectiveness of UV, Ozone, or Hydrogen Peroxide Vapor (HPV) room sterilization?

Bacterially lethal applications of UV, Ozone or HPV kill any bacteria on surfaces. Surface ATP tests don’t discern live or dead cells, so taking an ATP test immediately after sterilization of an unclean surface will always result in ATP presence. This doesn’t mean that you can’t use ATP testing if you also use UV, Ozone, or HPV room sterilization! Instructions for all of these technologies recommend thorough cleaning before application, so to ensure your sterilization is most effective, you should validate cleaning thoroughness before sterilization. ATP testing should be used before terminal sterilization to ensure cleaning has thoroughly prepared the room for effective sterilization with UV, Ozone, or HPV.

Can an ATP system be used to verify cleanliness of reusable items like endoscopes or surgical tools?

Yes. Hygiena provides high-sensitivity ATP detection swabs that are suitable for the sensitivity required by sterile services. To verify cleanliness of endoscopes, the EndoSwab can be used in conjunction with ATP test devices. 

Can an ATP system identify Methicillin-resistant Staphylococcus aureus (MRSA) or Clostridium difficile (C. diff) on a surface?

Not specifically. The only way to identify MRSA or C. diff is with microbiological testing methods that can take several days for results. ATP systems only detect the presence of organic materials on a surface and cannot detect specific strains of bacteria. Properly cleaned surfaces absent of ATP will not have the organic material required to facilitate harborage and growth of bacteria. 

How many ATP tests do I need to take to accurately monitor cleanliness in my hospital?

One misconception of ATP monitoring systems is that pre-cleaning and post-cleaning measurements need to be taken to measure cleaning effectiveness. ATP swabs should never be used on surfaces that are known to be soiled or dirty, as they will always result in a Fail test result. Sanitation companies and chemical suppliers will often use pre-clean and post-clean ATP tests to demonstrate the effectiveness of cleaning products, but this demonstration should not be confused with regular use in an ATP monitoring program. ATP monitoring should only be done after cleaning and not before and after cleaning. This is a key advantage of ATP monitoring over fluorescent marking systems that require multiple room visits as described above. Hospitals also do not need to monitor every single location available, but only a statistically representative sample in order to gather sufficient data for reporting and results interpretation. To estimate testing frequency for your hospital, and to calculate the ROI on cleaning performance, use the Testing Frequency & ROI Calculator found here. For detailed instructions on identifying test locations in your hospital, refer to the System Implementation Guide.

Why is ATP testing a better way to verify cleanliness?

Visual inspection is no longer a sufficient measure of cleanliness in healthcare facilities. In order to measure the cleanliness of surfaces and medical tools after cleaning, healthcare professionals around the globe are turning to ATP testing. The old way of cleaning verification required a supervisor to visit a room after a patient has been discharged to mark surfaces with a fluorescent gel pen. Then after environmental services staff completed cleaning, the supervisor would need to return to the room to perform a visual inspection of surfaces. This inspection was subjective, not measurable, required a supervisor to waste valuable time, and held up room turnover. With ATP testing, supervisors only need to visit a room once. Test swabs take only 15 seconds and can objectively quantify the cleanliness of a room. This allows rooms to be turned over faster, enables supervisors to get more work done, and holds cleaning staff accountable for proper cleaning with actual, quantifiable data. Learn more here.

How many healthcare facilities are using ATP monitoring?

A majority of US hospitals are measuring cleanliness with ATP monitoring systems. ATP systems are also prevalent in the UK and parts of Europe. The proportion of hospitals using ATP monitoring is growing and is expected to continue rising as hospitals continue to battle growing healthcare associated infection (HAI) rates and increasing demand for cleaner facilities.

What is an RLU?

RLU stands for Relative Light Unit and is the unit of measure used in bioluminescence. Luminometers measure and quantify light with an RLU output. Because manufacturers use different sensor technologies and algorithms for adding up the photons, RLU measurements will vary from system to system. However, because the ATP bioluminescence reaction is linear, the more ATP present means the more light will be present. (Comparing RLU values is like comparing Fahrenheit and Celsius; they are two different scales for the same temperature.) Using Hygiena’s SystemSURE Plus and UltraSnap ATP swabs: 1 RLU = 1 femtomole ATP (1×10 -15 mole)

For more information on comparing systems or if you are interested in learning about how to switch from one system to another, click here.

How does ATP monitoring work?

Hygiena ATP testing devices contain a natural enzyme found in fireflies. This enzyme produces a simple bioluminescence (light-producing) reaction when it comes into contact with ATP. Using bioluminescence technology, the SystemSURE Plus luminometer can measure extremely low levels of ATP collected with testing devices. Measuring the amount of bioluminescence from an ATP reaction provides an excellent indication of surface cleanliness because the quantity of light generated by the reaction is directly proportional to the amount of ATP present in the sample. The bioluminescence reaction is immediate so results can be processed at the testing site in seconds. Results are expressed numerically on the luminometer screen in Relative Light Units (RLU).

Bioluminescence is the result of a biochemical reaction and is the science behind Hygiena ATP detection tests. The reaction includes the following elements:

  • Luciferin /Luciferase -naturally occurring in fireflies
  • Adenosine triphosphate (ATP) – the energy molecule of all living organisms
  • Oxygen- a catalyst

Luciferin /Luciferase + ATP + O2 == == Light output

The reaction occurs in two steps:

  1.  The substrate combines with ATP and oxygen, which is controlled by the enzyme.
  2. The chemical energy in step 1 excites a specific molecule (the combination of Luciferin and Luciferase). The result is decay which is manifested as photon emission, or light production. The light is simply a byproduct of the chemical reaction and does not depend on light.

Are there any studies or white papers on ATP testing?

Download a full report of published studies featuring ATP testing here: ATP Cleaning Verification Published Study Summaries . Studies by the CDC and other non-governmental institutions have recommended ATP testing as way to verify cleaning of the healthcare environment and endoscope reprocessing. A 5-year study of two UK hospitals (North Tees and Hartlepool) using Hygiena’s system was released in December 2012. This study shows how using ATP monitoring improved hospital cleanliness and reduced C. diff infections more than 35%. ATP testing has been the industry standard for food and beverage processors for many decades. Silliker Laboratories, the leading international food testing and safety laboratory, conducted an independent study comparing five commercial ATP systems. This study found that Hygiena’s products offer superior linearity, sensitivity, repeatability, and accuracy.

For more information, or a copy of the full report, complete this form.