研究三種精子染色質分散 (SCD)測定在男性不孕症中精子 DNA 片段化的臨床應用
Investigating the Clinical Utility of Three Sperm Chromatin Dispersion (SCD) Assays for Sperm DNA Fragmentation in Male Infertility
Yuan-Chi Huang4, Hsin-Ta Lin1, Meng-Hsing Wu1, Li-Chung Tsai3, Yu-Ying Chen1, Kuei-Hsiang Hung1, Po-Hsuan Wu1, Ta-Sheng Chen1, Huang-Tz Ou3,4, Yu-Sheng Cheng4
1Department of Obstetrics and Gynecology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan
2Institute of Clinical Pharmacy and Pharmaceutical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan
3School of Pharmacy, College of Medicine, National Cheng Kung University, Tainan, Taiwan
4Department of Urology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan
This study aimed to assess the reproducibility, image quality and clinical utility of the three sperm chromatin dispersion (SCD) assays for accessing sperm DNA fragmentation, i.e., LensHooke R10®(R10), Halosperm G2®(G2), and BASO®(BA).
This study was proved by the Institutional Review Board of National Cheng Kung University Hospital (NCKUH), Tainan, Taiwan (A-ER-110-017). A total of fifty male participants were recruited, and all collected semen samples underwent semen analyses and SCD assays. The DFI was defined by the percentage of very small-size halo, without a halo, and degraded sperm cells. At least 500 sperm cells typically need to be evaluated in each specimen to produce the DFI. All sample slides obtained after chromatin staining followed each SCD kit procedure were observed under the 20X objective of Olympus BX53 under the bright-field light source. These target images were then captured and converted to highly contrasting images equal to the 100 X objective of the microscope. Intra- and inter-observer variability of DFI data from three different SCD measures were analyzed.
The G2 and R10 produced satisfactory variance coefficients (5.53%, 5.67%) compared to BA (14.8%). The DFI data from the R10 had lower intra-observer variability, in terms of higher intra-class coefficient (0.9615),than that of the G2 (0.8847) or BA (0.8824). Inter-observer variability of three SCD kits in scoring the DFI was comparable and satisfactory (concordance correlation coefficients ranging from 0.9895~0.9630). Five categories of SCD patterns processed by the R10 revealed the highest structured resolution in defining sperm core from halo width compared with G2 and BA.
This study provides the first preliminary evidence on the comparative reliability of the three available SCD kit assays in assessing sperm DNA fragmentation. The R10 was the most reliable SCD assay with better image quality to detect sperm DNA fragmentation than G2 and BA.
Keywords: sperm chromatin dispersion, DNA fragmentation index, male infertility.