Technical Release Bulletin [Link to document]
QC and proficiency testing of QwikCheck™ Vitality Kit
Compliance date: May 11, 2018
Authority: Marina Friling, Clinical manager Distribution: QwikCheck Vitality kit users
Purpose of this document:
To provide instructions for how to QC the QwikCheck Vitality product and test staff proficiency
Preparing and testing semen samples:
- Collect 1 fresh Human semen sample that meets the following criteria:
% Motility > 70%.
Volume > 1ml
- Wait for full liquefaction.
- Mix the semen sample well.
- Split the liquefied sample into two equal volume aliquots. Label Sample #1 and Sample #2.
- Sample #1: Keep at Room Temperature.
- Sample #2: Kill the cells by placing them into a standard freezer for 20 minutes, then allow sample to recover to Room Temp.
- Thoroughly mix each sample.
- Place 5 μl of each sample on a separate 1” x 3” lab slide.
- Add 5 μl of QwikCheck™ Vitality solution to each sample and mix with a pipette tip in a swirling motion.
- Cover the slides with a 22 mm x 22 mm coverslip and wait for the samples to stop drifting.
- Assess Vitality of each sample by counting 200 cells. Live spermatozoa have white or light pink heads and dead spermatozoa
have heads that are stained red or dark pink.
- NOTE: If the stain is limited to only a part of the neck region, and the rest of the head area is unstained, this is considered a
“leaky neck membrane”, not a sign of cell death and total membrane disintegration. These cells should be assessed as alive.
- Calculate the percent of vital cells from the total sperm counted.
- Repeat steps 8 through 13 with a second operator to confirm stain reaction and inter-operator variability.
QC Pass criteria:
PASS: % Vitality of Sample # 1 > % Vitality of Sample # 2.
FAIL: Deviation from PASS criteria, or no stained cells.
Proficiency Pass criteria:
Establish Allowable Target Range between operators:
Mean vitality value for sample #1 of both operators +/- 3 standard deviations or 25% whatever is greater
PASS: The vitality values of both operators fall inside the Allowable Target Range.