Bugcount® Fuel

These instructions are applicable to Bugcount® Fuel Test Kit | 3 tests and Bugcount® Fuel Test Kit | 25 tests.

To ensure that results obtained from Bugcount® Fuel testing are reliable, samples must be collected and handled in accordance with application-specific standards and guidance for microbiological sampling and testing.

Bugcount® Fuel is compliant with ASTM D7687 Standard Practice for Measurement of Cellular Adenosine Triphosphate in Fuel and Fuel Associated Water with Sample Concentration by Filtration.

General sampling guidance :

• Minimize delays between sampling and testing.
• Clean and disinfect area around sampling port’s external surfaces, work surface, and sampling device’s internal surfaces.
• Collect samples in previously unused, preferably sterile, sample containers.
• Personal protection equipment (e.g., nitrile gloves, safety glasses, etc.) is recommended to be worn for the entire procedure.
• Replace the chambers of your device twice a year using the Bugcount® One ATP Analyzer Maintenance Kit.

1. Capture – Isolate live cells

• Mix the sample well; remove the plunger from the 50mL syringe and attach a new syringe filter.
• Pour the recommended volume of sample into the syringe (Fuel: 20mL | Fuel-associated water: 5mL). Other sample volumes do impact the calculations, for more information, see the note on step 5.
• Use the plunger to slowly push the sample through the filter into a waste container.
• Detach the filter, then remove the plunger from the syringe.

2.2 Dry – Remove interferences

• Re-attach the filter to the syringe.
• Use the plunger to slowly push air through the syringe to dry the filter.
• Detach the filter, then remove the plunger from the syringe.

3. Release – Release ATP from the filter to get final sample extract

• Re-attach the filter to the syringe.
• Empty an UltraLyse 7 tube into the syringe
• Use the plunger to slowly push the UltraLyse 7 through the filter into a new UltraLute (dilution) tube.
• Cap the tube and invert 3 times to mix well. This sample is now ready for step 4.

4. Measure sample relative light units (RLU)

• Using a fresh exact-volume pipette (see TIP box for guidance), add the Luminase Enzyme solution to a new, clean test tube.
• Using a fresh exact-volume pipette, transfer the sample from Step 3 into the same test tube as the Luminase Enzyme solution, then swirl to mix.
• Place this test tube into the luminometer and take a reading.
• Record the result in Relay™, on the Bugcount® analyzer, or in the table provided as A cATP RLU.

5. Quantify ATP

Tip: Relay™ AND Bugcount analyzer can do the work For you

When all inputs are entered in Relay’s Run tests workflow, the results will calculate automatically. Select Save Test to catalog and share your results with your team. Or, run the test on the Bugcount analyzer and import the results to Relay for sharing.

Alternatively, you can manually calculate your test result using one of the simple calculations shown below corresponding to your sample type. This calculation will convert the RLU output of the luminometer to cellular ATP (cATP), which is reported in pg/mL as your final test results. If performing more than 3 tests, copy the table below into a spread sheet or notebook in order to keep results and calculations organized.

Select your equation based on sample type

\textit{Result (cATP)} =

{

\frac{

\colorbox{#72BF44}{\color{white}A}

-

\colorbox{#F15A24}{\color{white}B}

}{

\colorbox{#8E5BAA}{\color{white}C}

}}

\times 500
 

\textit{Result (cATP)} =

{

\frac{

\colorbox{#72BF44}{\color{white}A}

-

\colorbox{#F15A24}{\color{white}B}

}{

\colorbox{#8E5BAA}{\color{white}C}

}}

\times 2000
 

For sample volumes outside of those recommended in Step 1, use the following equation where V represents the actual sample volume filtered:

\mathrm{Result (cATP)}\
=
{

\frac{

\colorbox{#72BF44}{\color{white}A}

-

\colorbox{#F15A24}{\color{white}B}

}{

\colorbox{#8E5BAA}{\color{white}C}

}}
\times
\frac{10{,}000\ }{V_{\mathrm{sample}}\ \mathrm{(mL)}}

Troubleshooting

Additional Resources

myLuminultra:

• https://my.luminultra.com

Contact support:

• https://my.luminultra.com/hc/en-us/requests/new
• help@luminultra.com