UNIVERSITY HOSPITAL

AINTREE
DIALYSIS UNIT
WARD 20
PROTOCOL FOR
MICROBIOLOGICAL
SAMPLING FOR THE GAMBRO
AK 200 ULTRA S

CHRISTINE JONES
RENAL NURSE

INSTRUCTIONS FOR MICROBIOLOGICAL SAMPLING

A bacteriological control of the fluid after the two ultrafilters is to be
performed at the end of the filter life cycle, which normally means
once a month. If the filters are changed more frequently than
monthly, make sure to do the microbiological tests corresponding
to the filter changes.

SAMPLING POSITIONS
1. Post ultrafilter paper.
2. Infusion port (i.e. out of machine).
3. Water between RO and machine (i.e. into machine).

MATERIAL FOR SAMPLING.

1. Sterile filter holders (swinnex disc filter holder) equipped with
sterile 0.2um membrane filters for cultivation. The membrane
filter is put into the filter holder and then the holder is sterilised
preferably by autoclaving at 121oC for 20min. One filter holder
is used for each sampling position.
2. Sterile sampling tubes, two tubes for each sampling position.
3. Measuring cylinder with at least 1000ml volume.
4. Sterile syringes x 2 (20ml)
5. Sterile needle.

METHOD OF SAMPLING
Sampling needs to be done one week before the ultra filters are
due to be changed and results obtained before they are changed.
The bacteriological sampling can be done before or after a
treatment. Make sure that the machine is in HD, bicarbonate mode
and priming position.
A) Sampling through the swinnex filter holder.
1. Disconnect the hook functioning as a joint in the sampling
tubing.
2. Connect the swinnex filter holder to the tubing, the threaded
connector on the holder is the inlet.
3. Insert the pump segment into the blood pump and the tubing
into the arterial line clamp.

4. Discard the green cap on the remaining hook and place the
hook on the side of the measuring cylinder. The filter holder
must be kept upside down until the membrane filter is wetted to
remove air from the holder. The fluid will be collected in the
measuring cylinder.
5. Spray the infusion port with 70% ethanol and connect
aseptically the end of the sampling tubing to the infusion port.
6. Turn the three-way tap to allow passage of fluid from the port
only into the tubing.
7. Set the blood flow rate to 75ml/min.
8. Press the priming button.
9. Press the blood pump button.
10. Press the fluid bypass button.
11. When the filter in the holder is wetted, discard the hook with
which the holder hangs onto the measuring cylinder, turn the
holder upright and fix it on top of the cylinder.
12.When approx. 950ml has been collected, turn the valve on the
three-way tap to close on the infusion port so that only air
passes through the tubing and pump the fluid through the
sampling tubing.
13.When the tubing is empty, stop the blood pump and press the
fluid bypass button. Close the clamps either side of the swinnex
holder and cut the tubing outside the clamps. Remove the
tubing from the machine and discard.
14.Send the swinnex holder unopened for immediate culturing.
B) Sampling fluid pre and post filters
1. Spray the infusion port with 70% ethanol.
2. Connect aseptically a 20ml syringe to the free port on the threeway tap.
3. Turn the valve to close to the tubing, so that the passage of fluid
goes to the syringe and withdraw 20mls.
4. Put this fluid into two of the provided test tubes (i.e. 10mls in
each) and label as the OUT fluid.
5. Locate the length of tubing at the back of the machine from the
RO to the machine.
6. Spray the needle port with 70% ethanol.
7. Use a 20ml syringe with a green needle and remove 20mls via
this port.
8. Decant this fluid into the two remaining test tubes and label as
the IN fluid.
9. Send all four test tubes with the swinnex filter holder to the RLH
bacteriology dept. via taxi.

RESULTS
The results should conform to European Pharmacopoeia
standards.
For dialysis fluids and RO water this should be less than 0.25Eu/ml
For ultra-filtration output waters this should be less than 0.03Eu/ml.
If all results are within range then the ultrafilters may be changed.
If, however there is an abnormal result the machine must be
retested on the same ultrafilters, then put through chemical
disinfection and not used for online therapies.
If still abnormal change the ultrafilters more frequently and check
quality of plant room water.
If the result is normal on retest without any intervention please
contact Bob Graham at RLH bacteriology dept 706 4420.