ATP Testing Guide for Hygiene Efficacy

with Hygiena SystemSure Plus Luminometer

There are several testing methods that can be used to validate the efficacy of hand and footwear
hygiene methods. Each method has its limitations and should be fully evaluated before a system
of ongoing hygiene validation is written into an SOP or GMP system. This document explores our
recommended testing method: an Adenosine triphosphate (ATP) test.

What is ATP? FIGURE 1

ATP is an organic compound that provides energy to drive many

processes in living cells and organisms, such as muscle contraction
Adenosine triphosphate (ATP)
and chemical synthesis. ATP serves as a great marker for the chemical structure
prevalence of protein-like material on the surface of the skin (or
other organic surface)—some of which could be dangerous and
pathogenic.

What is ATP testing?

The ATP test is a process of quickly measuring actively growing
microorganisms through detection of Adenosine triphosphate, or
ATP. In ATP testing, microorganisms on the surface of the hands
combine with a certain enzyme to produce light which can then be
measured using a device called a Luminometer. The amount of light that is “read” by the device is expressed in Relative
Light Units (RLU’s). The more microorganisms that exist on the surface of the skin, the more light that is generated
and thus, the higher the RLU reading will be. High ATP levels indicate that more
organic material—some of which may possibly be harmful—are present. Organic
material on the surface of the hands may fall into one of two categories: transient
flora and resident flora.

Te r m ino lo gy

Resident flora: organisms that naturally occur on skin and do not cause disease.
Resident flora are essential to skin health and actually help protect the skin from
transient organisms.

Transient flora: Transient or temporary skin flora refers to the microorganisms

that transiently colonize the skin. This includes bacteria, fungi and viruses,
which reach the hands, for example, by direct skin-to-skin contact or indirectly
via objects.
How to conduct your
own ATP test

What you'll need

• Hygiena UltraSnap Swabs™
• Hygiena SystemSure Plus Luminometer™
• Facility to clean hands, or CleanTech® Automated Handwashing Station (recommended)
Note: While there are many ATP testing devices available on the market, we recommend using Hygiena SystemSure Plus Luminometer.

Instructions
1 2 3 4

Remove swab from tube and swab the palm Replace swab in the tube Select the test location Results will display in 15
of the dominant hand, applying sufficient and activate by bending in the Luminometer. seconds. Swab may be
pressure to create flex in the swab shaft. the bulb forward and Insert the swab into the discarded after use.
Rotate to collect samples on all sides of the backward. Squeeze to chamber and press "OK"
swab tip. expel liquid into the tube. to initiate measurement.
Shake for 5 seconds.

How to measure reduction: pre & post swabbing

An effective hygiene event results in reduced microorganisms on the hand. To measure this, make sure to capture a swab
of the natural hand first, and then conduct a second swab after a hand wash is completed to measure the difference.

Did you know?

You don't want to kill ALL the germs on your skin! The goal of
effective hand hygiene is to remove the transient organisms from
the surface of the skin with as little disruption and damage as
possible to the resident organisms, or the natural flora that the skin
needs to remain healthy.
Using ATP testing for process validation

Determining your methods

There are two basic ways to use ATP testing for hand hygiene validation:

1. Pass / Fail Limit Method: involves setting a certain RLU limit and measuring whether employees RLU counts fall
within this range.

2. Percentage Reduction Method: involves conducting a pre-hand wash swab and a post-hand wash swab to measure
the percentage reduction in RLU counts.

Protocol considerations for operators

The following considerations should be taken into account
when implementing an ATP testing protocol within a Why ATP testing
manufacturing or processing facility. is recommended
ATP readings will never reach 0. There will always be living
check Real time results. Forget the time
organisms and proteins on the skin which will result in RLU and hassle required of incubation or
readings. This does not mean that dangerous pathogens microbiological reading in a certified lab.
are present.
check It involves fewer variables compared to
Each individual in your facility will have different naturally- other test methods, increasing reliability
occurring levels of ATP, and some unique individuals will and accuracy.
have very high levels which will not be reduced below a check Far less expensive and requires fewer
certain level without damaging the skin. Therefore, you materials.
should always expect to find “outliers” in your test protocol,
check The incidence of outliers (data that
or individuals with unusually high levels of baseline ATP
cannot be explained) is much lower.
counts.

Lastly, it is very important that the same test methodology

and technique be used for every test subject, including
location of swab site, firmness of swab pressure, coverage of swab tip, and avoidance of cross contamination of swab. Every
individual tasked with conducting an ATP test should receive the same training, and the protocol itself must be validated.

Using the Percentage Reduction Method

We recommend the Percentage Reduction Method for testing with ATP Luminometers for two reasons. For one, this
method reduces the number of outliers due to naturally high levels of ATP. Second, focusing on a percentage of reduction
eliminates the need to establish what the pass/fail limit should be, which can differ for every population and facility.

Using the Pass/Fail Limit Method

The Pass / Fail Limit Method is much more sensitive to variability in testing and results in more outliers, or individuals who
fail to meet the minimum RLU reading standard. However, this method is frequently used as a training tool to demonstrate
to individuals that they are not washing consistently every time they wash and that their washing method may not be
effective—when, if compared to their baseline RLU counts, may not necessarily be true.
The challenge of pass/fail limits
In fact, most ATP test device companies have not developed a protocol or a pass/fail RLU limit for the testing of hand
or skin hygiene because it is nearly impossible to determine a standard limit that takes all environments, individual
characteristics, and processes into account. Those that do have a protocol typically recommend that a realistic pass/
fail limit be established which factors in the specific population to be tested as well as the type of hygiene that will be
occurring (e.g., surgical scrub, alcohol rub, healthcare hygiene, manual wash, soap type, automated hand wash, other).

A standard rule of thumb is to aim for under 100 RLUs after a regular 30--40 manual wash. However, if an individual is tested
and has a very high pre-wash RLU reading, they may still achieve a sufficiently high percentage reduction if compared
with the post-wash RLU reading. For example, Worker #4 in Figure 2 would fail in a process where the pass/fail limit is
100 RLUs, however when you compare this person's post-wash RLU count to their pre-wash RLU count, the reduction
percentage is significant (91%). Individuals that have naturally-occurring high RLU counts may not be able to achieve
an RLU reading under 100 without suffering
damage to the skin. Some individuals with
high readings should be tested multiple
times to establish their individual pass/fail
level.

Setting your targets

One way for a facility to establish its own
baseline percentage reduction goal and/
or its own Pass/Fail RLU limit is to perform
a 10 subject ATP test protocol and use the
average percentage reduction found among
the 10 test subjects when comparing pre-
wash RLUs to post-wash results. See below
the 14 subject test conducted by Meritech as FIGURE 2 A comparison of percentage reduction
an example. and pass/fail limit results.This testing was conducted
using Hygiene SystemSure Luminometer.
Disclaimer: These recommendations and data are based
on using Hygiena SystemSure Luminometer. Other ATP
Testing devices may produce different results.

Need help with

your protocol?
Our team of hygiene
experts is ready to help!

www.meritech.com | 800.932.7707
WWW.MERITECH.COM | 800-932-7707