计算机辅助精子分析室内质控物的研究与应用

Internal quality control products for computer-assisted sperm analysis: Research and application

目的:通过自制精液质控品用于计算机辅助精子分析(CASA)的日常室内质控,以提高精液检测的室内质控质量,获得良好的精密度及准确度,提高为兵服务质量。方法:依次使用高、低精子浓度水平的精子商品化质控品校准CASA,同时将15份精液混合质控品,分装在75个冻存管内,通过标准程序用CASA进行精子浓度、活力和运动参数(VCL,VSL,VAP,LIN,WOB,STR)的分析。连续监测50天。为了分析冷冻精子做为室内质控品对其浓度准确性的影响,再用Makler板进行计数。结果:高低浓度水平商品化精液质控品的变异系数CV分别为6.18%和7.85%。而自制质控品的浓度为(25.97±1.41)×10^6/ml,CV值为5.42%。精子的活动力(PR+NP)%及运动参数VCL,VSL,VAP,LIN,WOB,STR值分别为(22.15±1.75)%、(59.18±2.05)μm/s、(26.79±1.2)μm/s、(34.98±1.4)μm/s、(46.81±1.55)、(60.52±1.3)、(76.46±1.98)变异系数CV分别为7.9%、3.46%、4.48%、4.01%、3.3%、2.15%、2.59%。同时利用Makler板计数自制精子的浓度和活力值分别为(34.39±2.37)×10^6/ml和(38.04±1.69)%,CV值分别为6.89%和4.44%。通过平均值及标准差画出每一项指标的Levey-Jennings质控图。结论:用液氮冻存精液的方法制备自制室内质控品,来监测CASA所检测精子浓度,精子运动参数的准确度及精密度的方法是可行的。且自制质控品检测精子浓度比商品化质控品所测的变异系数要小。

Objective: To investigate the application of the self-made semen quality control(QC) product in internal QC of computer-assisted sperm analysis(CASA). Methods: CASA was calibrated with high-and low-concentration commercially available semen QC product and meanwhile 15 samples of self-made mixed semen QC product were placed in 75 cryotubes containing liquid nitrogen, followed by CASA of the concentration, motility, curvilinear velocity(VCL), straight line velocity(VSL), average path velocity(VAP), linearity(LIN), wobble(WOB) and straightness(STR) of the sperm using standard procedures and 50 days of continuous monitoring. The Makler counting plate was used to measure the concentration and motility of the self-made sperm. Results: The coefficients of variation(CV) of the commercially available semen QC product at high and low concentrations were 6.18% and 7.85%, respectively. CASA showed that the concentration of the self-made QC product was(25.97 ± 1.41) ×10^6/ml, with a CV of 5.42%, and the sperm motility, VCL, VSL, VAP, LIN, WOB and STR were(22.15 ± 1.75)%(CV = 7.9%),(59.18 ± 2.05) μm/s(CV = 3.46%),(26.79 ± 1.2) μm/s(CV = 4.48%),(34.98 ± 1.4) μm/s(CV = 4.01%), 46.81 ± 1.55(CV = 3.3%), 60.52 ± 1.3(CV = 2.15%) and 76.46 ± 1.98(CV = 2.59%), respectively. The concentration and motility of the self-made sperm detected with the Makler counting plate were(34.39 ± 2.37) ×10^6/ml(CV = 6.89%) and(38.04 ± 1.69)%(CV = 4.44%), respectively. Levey-Jennings QC charts were plotted for the indicators using the means and standard deviation. Conclusion: The self-made internal QC product by liquid nitrogen cryopreservation is feasible and effective for monitoring the accuracy and precision of CASA-derived sperm concentration and motion parameters, and it has a smaller CV than the commercially available QC product in measuring sperm concentration. Natl J Androl, 2020, 26(10): 895-899.