P–009 A modified sperm chromatin dispersion test, LensHooke® R10, for quick and accurate determination of human sperm DNA fragmentation

Study question The performance and efficiency of the LensHooke® R10 test kit were evaluated by the clinical examination for precision, accuracy, and time. Summary answer The LensHooke® R10 based on sperm chromatin dispersion test offers not only quick testing for sperm DNA fragmentation but also reliable and accurate test results. What is known already Sperm chromatin dispersion (SCD) test, one of the most commonly used testing for sperm DNA fragmentation (SDF), can be conducted promptly and without the need for expensive laboratory instruments. However, the main disadvantage of the SCD test is inter-observer variability in categorizing the size of characteristics halos surrounding the core of sperm. Moreover, it takes more than one hour to accomplish whole assay procedures making this testing an inefficient diagnostic tool. These may hinder its broad availability among andrology laboratories or prevent it from being routinely used for the evaluation of male infertility. Study design, size, duration A total of 108 participants was included in this prospective study. Data was collected from the reproductive medicine center between June and December 2020. Participants/materials, setting, methods This study included 108 consecutive male partners of couples attending for assisted reproductive treatment. SDF was simultaneously tested by using LensHooke® R10 (R10) and Halosperm® G2 (G2) respectively. We evaluated the correlation and agreement between two SCD-based test kits. The repeatability and reproducibility of the SCD kits were assessed by intra-and inter-observer agreement experiments. The sensitivity, specificity, positive predictive value, negative predictive value for the R10 was determined by receiver operator characteristics (ROC) curve analysis. Main results and the role of chance The R10 produced more clear sperm core and dispersed chromatin, therefore highly recognizable images can be easily and accurately categorized when scoring of SDF. It took 50% less time for SDF testing by the R10 compared to the G2 (38.26 ± 9.85 minutes vs. 76.52 ± 19.7 minutes, P < 0.0001). The SDF% results showed a strong correlation for the R10 and G2 with Spearman’s coefficients of rank correlation (rho) above 0.8 (P < 0.0001, N = 108). The R10 showed 89.8% accuracy with 87.9% sensitivity, 90.8% specificity, 82.9% PPV, and 93.7% NPV on the measurement of SDF% at the threshold value of 22%. Intraclass correlation coefficients (ICC) >0.9 showed a strong agreement between two observers on the testing of SDF using the R10. ICC >0.9 showed a high intra-observer agreement within 4 repeated testing on SDF using the R10. The R10 showed an intra-observer’s precision of coefficient variation, CV < 10% for SDF%. In addition, SDF% test results obtained by the R10 for asthenospermic (31.8% ± 16.7%), teratospermic (22.9% ± 14.4%), and oligoasthenoteratozoospermic samples (36.6% ± 14.4%) were significantly higher than that observed in normozoospermic samples (15.3% ± 10.2%, p < 0.05), was comparable with the G2. Limitations, reasons for caution The sample size of 4 semen specimens used to evaluate the intra-and inter-observer agreement was a limitation. Besides, evaluating the relationship between the SDF and clinical outcome of ART is necessary for further study. Wider implications of the findings: The new in vitro diagnostics reagent, LensHooke® R10, is a simple and quick test kit that offers reliable and accurate test results of sperm DNA fragmentation, can be routinely used in male infertility evaluation. Trial registration number CS2–20012

Sperm DNA Fragmentation Among Smokers and Non-Smokers as Measured by Sperm Chromatin Dispersion Test

Introduction: Traditionally, seminal fluid analysis is done to assess male infertility particularly sperm concentration, morphology and motility. Sperm chromatin dispersion test explores the quality of the sperm DNA structures. High sperm DNA fragmentation is associated with unexplained infertility, failure of assisted reproduction and recurrent miscarriages. Smoking has been associated with high sperm DNA fragmentation in some studies but not in others. Materials and Methods: A comparative cross-sectional study, involving 32 smokers and 32 non-smokers to determine the association between cigarette smoking and sperm DNA fragmentation. Semen samples were collected from patients undergoing seminal fluid analysis (SFA) at the IIUM Fertility Centre and IIUM Medical Centre O&G Clinic from January 2017 to June 2018. Sperm chromatin dispersion test was done and level of 15% is considered upper limit of normal. Results: 53.1% in the smoker group have abnormal level of sperm DNA fragmentation compared to 34.4% of nonsmokers. The median and interquartile range for smokers were 15.30 and 19.0 (10.425 - 29.375) respectively while for non-smokers were 8.25 and 17.8 (4.075 - 21.850) which was statistically significant (p= 0.012). It was also noted that three participants in the smoker group (9.4%) has very high sperm DNA fragmentation index of more than 80% even though they have normal seminal fluid analysis. Conclusion: There is an association between smoking and high sperm DNA fragmentation. There is a trend of increased in DNA fragmentation in smokers even though their SFA results were normal.