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Human Reproduction, Vol. 14, No. 8, 2036-2040, August 1999
© 1999 European Society of Human Reproduction and Embryology

Intra- and inter-laboratory variability in the assessment of sperm morphology by strict criteria: impact of semen preparation, staining techniques and manual versus computerized analysis

Gerardo Barroso1, Ramazan Mercan1, Kemal Ozgur1, Mahmood Morshedi1, Paul Kolm2, Kevin Coetzee3, Thinus Kruger3 and Sergio Oehninger1,4

1 Department of Obstetrics and Gynecology, 2 Bio-statistics, The Jones Institute for Reproductive Medicine, Eastern Virginia Medical School, 601 Colley Avenue, Norfolk, Virginia, 23507, USA and 3 Reproductive Biology Unit, Department of Obstetrics and Gynecology, Tygerberg Hospital, University of Stellenbosch, Capetown, South Africa

We designed prospective studies to compare manual and computerized analysis of sperm morphology by strict criteria using different semen processing and staining techniques. A total of 54 semen samples were studied; slides were prepared from each subject from liquefied semen and after washing, and stained with Diff-Quik® or Papanicolaou. An intra-laboratory, blind assessment was performed manually (two observers) and using a computerized analyser (two readings). This demonstrated a very good correlation between manual analysis of liquefied and washed samples with both staining techniques [intraclass coefficient (ICC) = 0.93 and 0.83]. Greater agreement was observed between computerized readings (washed samples) of Diff-Quik® (ICC = 0.93) than of Papanicolaou-stained slides (ICC = 0.66). An excellent intra-laboratory correlation was observed for within-computer readings (ICC = 0.93). There was moderate agreement between inter-laboratory computer readings (two centres, ICC = 0.72). Although there was lower inter-laboratory agreement for manual and manual versus computer readings, overall results of all manual and computer analyses showed good agreement (ICC = 0.73). Diff-Quik® staining is reliable for both manual (liquefied) and computer (washed) analysis of strict sperm morphology. Intra- and inter-computer analyses using this method reached satisfactory levels of agreement. There is still high inter-laboratory variability for the manual method.

Key words: inter-laboratory variability/intra-laboratory variability/normal sperm morphology/staining/strict criteria

4 To whom correspondence should be addressed


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