Zrimsek P, Kunc J, Kosec M, Mrkun J
Clinic for Reproduction and Horses, Veterinary Faculty, University of Ljubljana, Gerbiceva 60, 1000 Ljubljana, Slovenia.
Int J Androl. 2004 Feb;27(1):57-62. doi: 10.1046/j.0105-6263.2003.00448.x.
The resazurin reduction assay depends on the ability of metabolically active cells to reduce the resazurin redox dye to resorufin. In the present study we applied and made a diagnostic evaluation of a spectrophotometric application of the resazurin reduction assay to assess the colour change of resazurin reduction in butanol extracted colour to evaluate boar semen quality. Forty-one samples of boar semen from various breeds were included in the study. The absorption peaks for resazurin and resorufin were found to be 610 and 575 nm, respectively. Absorbance at 610 nm, where the minimum overlap of the two peaks was observed, was used in further analysis. Spearman rank correlation analysis was used to determine the correlation between the resazurin reduction assay and various semen parameters. The highest correlations were observed with the concentration of motile spermatozoa (r = -0.841; p < 0.001), sperm concentration (r = -0.833; p < 0.001), sperm index (-0.826; p < 0.001) and concentration of viable spermatozoa (r = -0.763; p < 0.001). Sensitivity and specificity, at 94.1 and 91.7%, respectively, indicate that the present test is highly accurate in discriminating between the samples according to the sperm index. When motile sperm concentration was used to distinguish between good and poor samples, high sensitivity (93.6%) was also found, whereas the test was only moderately, 80%, specific. The stability of butanol extracts in terms of A610 at different times of measurement confirmed that the resazurin reduction could be spectrophotometrically measured within 7 days from the time of assay performance, making the assay much more useful. Based on these results, the assay could be used as an additional tool for evaluating the quality of boar semen.
刃天青还原试验依赖于代谢活跃细胞将刃天青氧化还原染料还原为试卤灵的能力。在本研究中，我们应用了刃天青还原试验的分光光度法并对其进行诊断评估，以评估丁醇提取物中刃天青还原的颜色变化，从而评价公猪精液质量。该研究纳入了41份来自不同品种的公猪精液样本。发现刃天青和试卤灵的吸收峰分别为610和575nm。在观察到两个峰最小重叠的610nm处的吸光度用于进一步分析。采用Spearman等级相关分析来确定刃天青还原试验与各种精液参数之间的相关性。观察到与活动精子浓度（r = -0.841；p < 0.001）、精子浓度（r = -0.833；p < 0.001）、精子指数（-0.826；p < 0.001）和活精子浓度（r = -0.763；p < 0.001）的相关性最高。敏感性和特异性分别为94.1%和91.7%，表明本试验在根据精子指数区分样本方面具有很高的准确性。当用活动精子浓度区分优质和劣质样本时，也发现了高敏感性（93.6%），而该试验的特异性仅为中等水平，为80%。在不同测量时间下丁醇提取物在A610方面的稳定性证实，刃天青还原可在测定执行后7天内通过分光光度法进行测量，这使得该试验更有用。基于这些结果，该试验可作为评估公猪精液质量的一种额外工具。
