Critical evaluation of two models of flow cytometers for the assessment of sperm DNA fragmentation: an appeal for performance verification

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dc.contributor.authorSharma, Rakesh
dc.contributor.authorGupta, Sajal
dc.contributor.authorHenkel, Ralf
dc.date.accessioned2023-06-06T12:11:38Z
dc.date.available2023-06-06T12:11:38Z
dc.date.issued2019
dc.description.abstractLack of standardized, reproducible protocols and reference values is among the challenges faced when using new or upgraded versions of instruments in reproductive laboratories and flow cytometry. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay combined with flow cytometry routinely used for diagnostic measurement of sperm DNA fragmentation (SDF) is a unique example. Any change in the setting of the standard instrument, including upgrades of hardware or software, can lead to different results and may affect clinicians’ decision for treatment. Therefore, we compared TUNEL results of SDF obtained from a standard (C6) flow cytometer with a newer version of the same instrument (C6 Plus) and examined the cutoff, sensitivity, and specificity without calibration (adjustment) and after adjustment. Identical sperm preparation and matched acquisition settings were used to examine the performance of two flow cytometers. The strength of agreement of the results between the two observers was also assessed. After adjustment of the settings, overall concordance became high and the two cytometers showed 100% positive and negative predictive value with 100% area under the curve. The overall correlation coefficient observed between C6 and C6 Plus was highly significant (P < 0.0001; r = 0.992; 95% confidence interval [CI]: 0.982–0.997). After adjustment, the two cytometers showed very high precision of 98% and accuracy of >99%. The interobserver agreement on C6 flow cytometer for the two observers was 0.801 ± 0.062 and 0.746 ± 0.044 for C6 Plus. We demonstrated a strong agreement between the samples tested on the two flow cytometers after calibration and established the robustness of both instruments.en_US
dc.identifier.citationSharma, R. et al. (2019). Critical evaluation of two models of flow cytometers for the assessment of sperm DNA fragmentation: an appeal for performance verification. Asian Journal of Andrology, 21 (5) , 438-444. 10.4103/aja.aja_109_18en_US
dc.identifier.issn1745-7262
dc.identifier.uri10.4103/aja.aja_109_18
dc.identifier.urihttp://hdl.handle.net/10566/9039
dc.language.isoenen_US
dc.publisherMedknow Publicationsen_US
dc.subjectDNA fragmentationen_US
dc.subjectBioscienceen_US
dc.subjectSpermen_US
dc.subjectOxidative stressen_US
dc.titleCritical evaluation of two models of flow cytometers for the assessment of sperm DNA fragmentation: an appeal for performance verificationen_US
dc.typeArticleen_US

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