Hum. Reprod. Advance Access originally published online on April 21, 2005
Human Reproduction 2005 20(8):2140-2152; doi:10.1093/humrep/dei033
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Frequency of aneuploidy in sperm from patients with extremely severe male factor infertility
1 S.I.S.Me.R. Reproductive Medicine Unit, via Mazzini 12, 40138 Bologna, 2 Dept Ob/Gyn, S. Martino Hospital, University of Genoa, Largo Rosanna Benzi 10, 16132 Genoa and 3 Dept Ob/Gyn, University of Pisa, via Roma 67, 56126 Pisa, Italy
4 To whom correspondence should be addressed. Email: sismer{at}sismer.it
BACKGROUND: A protocol for the chromosomal analysis of sperm samples with a severely reduced number of sperm cells was designed. METHODS: A severe male factor condition was the main cause of infertility for 38 couples: 27 were oligoasthenoteratospermic (OAT) and 11 with non-obstructive azoospermia underwent testicular sperm extraction (TESE). A two-round fluorescence in situ hybridization (FISH) protocol was performed with probes specific for the chromosomes X, Y, 13, 15, 16, 17, 18, 21 and 22. The recording of the position of each sperm cell at the microscope allowed diagnosis of each spermatozoon for the nine tested chromosomes. RESULTS: A mean number of 122±78.5 sperm were diagnosed per patient with an incidence of total abnormalities corresponding to 13.4%. 
2-tests for the observed frequencies and goodness-of-fit test were highly significant in all cases. A significantly higher proportion of total aneuploidy was detected in 79% of the tested samples compared to the normal population. Testicular sperm were significantly more prone to aneuploidy than ejaculated sperm. CONCLUSIONS: The designed FISH protocol for the analysis of severe OAT and TESE sperm samples is reliable, implying that the studied sample is representative of the original population. In view of the high incidence of aneuploidy in most severe OAT and TESE sperm, the FISH analysis of pathological sperm samples can be routinely performed in order to estimate the chances of the paternal contribution to aneuploidy in the resulting embryos.
Key words: aneuploidy/disomy/fluorescence in situ hybridization/male factor infertility/preimplantation genetic diagnosis
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