Scoring human sperm morphology using Testsimplets and Diff Quik slides

For assessing sperm morphology, Testsimplets gives a statistically significantly lower number of normal forms, so its clinical application is not recommended over Diff-Quik, the quickest recommended technique.

Ilaria Natali, Ph.D., Monica Muratori, Ph.D., Valentina Sarli, B.S., Mario Vannuccini, Ph.D., Sonia Cipriani, Sc.D., Laura Niccoli, M.D., Claudia Giachini, Ph.D.

Volume 99, Issue 5, Pages 1227-1232.e2, April 2013


To compare two staining methods to assess sperm morphology: the quickest recommended technique (Diff-Quik, DQ) and Testsimplets® (TS), a technique that uses prestained slides and that is quite popular in In Vitro Fertilization (IVF) centers.

Prospective study.

Patients at the Sterility Center of the Obstetrics and Gynecology Unit of the Hospital of S.S. Cosma and Damiano (Azienda USL 3 of Pistoia, Italy).

One hundred four randomly enrolled patients attending the seminology laboratory.


Main Outcome Measure(s):
Statistical comparison of sperm morphology results obtained after staining of semen samples both with DQ and TS.

Our data show that TS gives a significantly lower number of normal forms than DQ [median: 6% (range 0-29%) vs. 12% (range 0-40%), respectively] as well as an overestimation of sperm head defects [median: 92.0% (range 67-100%) vs. 82.3% (range 55-100%), respectively].

The two staining methods should not be considered equivalent. Specifically, the lower reference limit established by the World Health Organization is not appropriate when sperm morphology is assessed by TS. The routine application of the TS for the evaluation of sperm morphology is therefore not recommended since it would lead to an overestimation of patients with a sperm morphology value below the lower reference limit (4%), thus potentially influencing clinical decisions.

  • The authors demonstrate the stark difference in morphology between DQ and TS, with DQ being more accurate in characterizing morphology. This is based on prior validation studies between DQ and Papanicolaou staining method. Papinicolaou staining is time consuming, more expensive, and very dependent on the experience of the clinician.

    There is always a role for finding a more efficient and cost effective method for diagnostic testing. However, if it comes at the expense of accuracy, it should not be used, especially when it has significant clinical implications (i.e. To proceed with IVF/ICSI vs. IUI vs. natural conception). We need to be cautious when developing new testing methods. If it does not perform at least to the current standards, it should not be utilized for clinical decision making of this magnitude.

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