Helping Hospitals Define Clean

Frequently Asked Questions



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 ATP Products

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.

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

Biolum reaction

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.


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 (1x10 -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.


graph 1 passcautionfail


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.


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 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.


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. 


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 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.


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 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.


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

Below is an image of the luminometer screen:

 Anatomy of Screen HC

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 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.



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.

 Clean test Sanitize

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.


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.


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. 


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.


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 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.

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).


Correct         Incorrect
proper swabbing   wrong swabbing

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 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.


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 do I contact Hygiena to purchase an ATP solution?

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

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 Monitoring Systems

EnSURE Touch, SystemSURE Plus 

Does the system need to be calibrated?

Hygiena ATP systems self-calibrate at start up to ensure accurate readings. To further verify calibration, we offer a Calibration Control Kit. Though the kit is not necessary for the performance of the system, a documented calibration process shows due diligence to auditors. With advanced technology, Hygiena systems will stay in calibration for the life of the device. 


Are there maintenance costs I should be worried about?

Unlike other systems on the market, Hygiena’s SystemSURE Plus does not require expensive yearly maintenance or service contracts. Monitoring systems come with a 1 year warranty. Should your system become damaged or non-operational, Hygiena will promptly replace your device with a loaner so that you do not experience any interruption in your testing program.


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  Surface Residue Tests


Do I need to refrigerate Medi-Check?

No. Medi-Check may be stored at room temperatures. The test devices are shipped on ice to compensate for high temperatures in transit. Upon receipt, the swabs may be moved to room temperature storage for the shelf life of the product.


What does it mean if the reagent color changes after the time recommended by the instructions?

Any color change should be disregarded after the recommended time frame. For MediCheck, any color change after 30 minutes should be ignored.


The reagent turned grey. What does this mean?

Any color change within the incubation time  (37°C for 30 minutes or 55°C for 15 minutes) indicates the presence of proteins. A grey color indicates a very small amount of protein was detected. For critical areas or high-risk products, a grey color should prompt a reclean and retest. For low risk areas or products, this should be considered a cautionary result. 


What is the difference between MediCheck and ATP tests?

MediCheck detects the presence of proteins only. ATP is the energy molecule of all organic material, whether it is bacteria, biological matter, or any other organic residue. Thus, ATP tests detect a wider range of residues, and also give results in 15 seconds. Medi-Check has a 15-30 minute incubation step in the test process.


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  SureTrend Software


Where can I download SureTrend software?

Go here: SureTrend Software Download Information

I have an old version of SureTrend software (3.0 or earlier). How can I update to the latest version?

Find details here: SureTrend Software Download Information


What OS is recommended for Hygiena's SureTrend Software?

See specifications here: SureTrend Software Download Information


Can I run SureTrend on a tablet?

Yes, as long as the tablet meets the hardware and software requirements listed above in "What OS is recommended for Hygiena's SureTrend Software?".
IMPORTANT: Windows RT tablets are not supported. 

Do I need to dedicate a database server to use SureTrend Version 4.0?

SureTrend Version 4.0 uses Microsoft SQL Server to store its data. By default SureTrend will install and use the Microsoft SQL Compact 3.5 edition for single and multi-user installations. You can upgrade the database to SQL Server Express and above, and then configure the SureTrend client to use your SQL server.


Is there a version of SureTrend for Mac?

No, SureTrend was designed for Windows, but you can run SureTrend on a Mac using software like Parallels or other virtualization software. The important requirement for the virtual machine is USB support. Some older versions of Parallels, VMWare, Microsoft Virtual PC do not support USB, so it is important to have a current version. If SureTrend is installed on a terminal server or if the user is remoting to the virtual machine, there may be issues with USB redirection.


Does SureTrend support Virtual Desktop or RDP?

SureTrend can run on a Windows Terminal Servier (RDS), Citrix, or other VNC software that will remote the COM port from the server to the desktop. Since SureTrend is running on the server, but the unit is connected to the desktop COM port, communication to the Unit is directed over the network. If your network latency is too high you may get COM port communication failures that require you to repeat the synchronization process with the Unit. Also, the SureTrend feature that automatically detects the COM port will not work and user must know the COM port used on the desktop.


Does SureTrend support virtualization such as Windows XP mode or VMWare?

SureTrend will run on virtualized version of Windows supported by SureTrend as long as the virtualization software supports COM ports or USB. Some users are running Windows 7 or 8 on their desktop computer, but spend most of their time in a Windows XP mode session to support other legacy applications in their organization. Having SureTrend run in the Windows XP mode helps the user stay in the Windows XP mode and not have to switch to the native desktop just to run SureTrend.


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 Other Questions


How can I get a Certificate of Compliance for a particular product or lot?

To download a Certificate of Compliance for a product, use the handy Certificate Search tool here

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