Standard Operating Procedures Clinical Protocol VII: Urine Sample Collection

** NOTE: The following procedure is to be performed wearing laboratory coat, gloves, eye protection, and mask.

PRINCIPLE

Sterile urine will be collected from patients at the indicated time points following randomization.

The collection of urine should be obtained from an indwelling urinary catheter.

Specimen Collection & Handling

1.Label all 4 x 2mL cryogenic freezing tubes per sample per patient.

2.Approximately 15mL of sterile urine is to be collected from the side port of each patient’s indwelling urinary catheter and placed into a sterile 50mL polypropylene tube.

3.In the laboratory, the urine is centrifuged at 1500 x g for 5 minutes with brakes off, in a horizontal, swinging bucket centrifuge.

4.Urine is then aliquoted into the labeled 2mL cryogenic freezing tubes using sterile techniques.

5.Urine sediment is discarded.

6.The aliquot are stored at ≤ –70°C until processing.

Supplies

•2mL cryogenic freezing tubes

•15mL sterile polypropylene conical tubes (Falcon/Becton-Dickinson, #35-2097)

•Sterile, endotoxin-free 10mL serological pipets (VWR, #53283- 740)

•Rubber bulbs for pasteur pipets (VWR, #56311-062)

•P1000 pre-sterilized, pipet tips (VWR, # 53508-830)

Equipment

•Clinical Centrifuge with swinging-bucket rotor. Must accommodate 15mL conical tubes [Put some dry cotton or gauge at the bottom of the 15mL tube holders in order to avoid sticking of CPT (glass) tubes].

•Ultra low temperature -70°C freezer