P11 Sperm aneuploidy: When to stop counting?

Tempest, Helen G.; Gillott, David J.; Grigorova, M.; Handyside, Alan H.; Thornhill, A.

doi:10.1016/s1472-6483(10)62327-9

Cited by 1 publication

(2 citation statements)

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“…Only clearly defined sperm nuclei with a flagella and containing at least one spot were analyzed and two signals of the same color were considered as different if they were the same size and were separated at least by a spot diameter. Since, the whole research protocol had been designed before data of Tempest and coll [13] were published, approximately 5000 cells were counted per sample, one half automatically, the other half manually. We did not modify the design because it also allowed us to detect statistical differences in each group (more than 1000 in the manual and in the automated group, and also between the two operators in the manual group).…”

Section: Methodsmentioning

confidence: 99%

“…This procedure, when performed manually by a trained technician is highly time consuming. Tempest and collaborators demonstrated that no difference is observed in aneuploidy rates when scoring 1000 cells or 5000 cells [13]. We evaluated several years ago, that adhering to strict scoring criteria and depending on the cell density on the slides, 3 to 15 hours are needed to enumerate the chromosomal content of 5.000 cells for a set of three probes.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

FISH and tips: a large scale analysis of automated versus manual scoring for sperm aneuploidy detection

Martinez

Gillois

Mitouard

et al. 2013

Basic Clin. Androl.

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BackgroundApproximately 1% of the spermatozoa found in ejaculate of healthy men are aneuploid and this rate increases in the population of subfertile and infertile men. Moreover, fertilization with these aneuploid sperm can lead to impaired embryo development. Fluorescent In Situ Hybridization (FISH) is the common cytogenetic tool used for aneuploidy screening on sperm. However, it is a time-consuming technique and cytogenetic or in vitro fertilization laboratories cannot routinely use it and face the increasing demand of such analyses before Assisted Reproductive Techniques (ART). As automation can be a clue for routine practice, this study compares manual and automated scoring of sperm aneuploidy rates using a Metafer Metasystems® device. The results obtained also contribute to global data about FISH on sperm cells.MethodsWe recruited 100 men addressed for sperm cryopreservation. They all signed an informed consent to participate in the study. 29 men were donors or consulted before vasectomy (control group) and 71 were suffering of Hodgkin’s disease or non Hodgkin lymphoma (patient group). One semen sample was collected for each patient, analyzed according to WHO criteria and prepared for a triple-color FISH using centromeric probes for chromosomes 18, X and Y. Automated scoring was performed using a Metafer Metasystems® device.Results507,019 cells were scored. We found a strong concordance between the automated and the manual reading (d < 0.01 in Bland-Altman test). We also did not find a statistically significant difference between the automated and the manual reading using Wilcoxon test for total aneuploidy rate (p = 0.06), sex chromosomes disomy (p = 0.33), chromosome 18 disomy (p = 0.39) and diploidy (p = 0.21). Cumulative rate of total aneuploidy was 0.78% ± 0.212% for patient group and 0.54% ± 0.15 for control group and among this, sex chromosome XY disomy rate was of 0.54% for patient group and 0.27% for control group.ConclusionThis study validates the automated reading for FISH on sperm with a Metafer Metasystems® device and allows its use in a laboratory routine.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%