title: 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.
abstract: 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.
authors.affiliation: Department of Obstetrics and Gynecology, Bio-statistics, The Jones Institute for Reproductive Medicine, Eastern Virginia Medical School, 601 Colley Avenue, Norfolk, Virginia 23507, USA.
authors.forename: G
authors.initials: G
authors.lastname: Barroso
authors.forename: R
authors.initials: R
authors.lastname: Mercan
authors.forename: K
authors.initials: K
authors.lastname: Ozgur
authors.forename: M
authors.initials: M
authors.lastname: Morshedi
authors.forename: P
authors.initials: P
authors.lastname: Kolm
authors.forename: K
authors.initials: K
authors.lastname: Coetzee
authors.forename: T
authors.initials: T
authors.lastname: Kruger
authors.forename: S
authors.initials: S
authors.lastname: Oehninger
mesh_terms: D000328:Adult; D002469:Cell Separation / Q000592:standards; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008297:Male; D011446:Prospective Studies; D012015:Reference Standards; D013094:Spermatozoa / Q000166:cytology*; D013194:Staining and Labeling