摘要
目的 探讨流式细胞术在精子染色质结构分析 (SCSA)方法对实验结果的影响。方法利用吖啶橙的异染性和流式细胞术检测精子染色质DNA对原位酸诱导变性的敏感性。结果 存放精子的方法不同 ( 4℃保存、稀释冻存、原浆冻存 )对COMPαt、αt、SDαt值均影响较大 ,稀释后液氮保存法的人为损伤最低。冻存精液融化后立即检测以及测量流速 <3 0 0个精子 /s对结果无影响。该法的批内、批间变异系数分别为 7 2 8%和 8 92 %。 511名健康男性的COMPαt平均值及标准偏差为( 8 7± 11 0 ) %。由于分析资料呈右偏态分布 ,COMPαt的参考值范围是 <2 6 8%。COMPαt与精液常规分析中的d级精子率、平均移动角度、精子密度中度正相关 ,与a级精子率、精子存活率分别呈中度负相关。结论 流式细胞术对精子染色质结构分析能快速分析精子DNA损伤 ,为评估精子亚临床损伤。
Objective To study the factors effecting on flow cytometric sperm chromatin structure assay.Methods The SCSA explores the metachromatic properties of acridine orange (AO) and flow cytometry to monitor the susceptibility of sperm chromatin DNA to acid induced denaturation in situ because of the low pH treatment. Results COMPαt, αt, SDαt value were different upon on the storage methods (①sperm was preserved in 4℃, ②sperm was cryopreservated after dilution ③sperm was cryopreservated). Results show that the second method above made the least artificial injury to sperm DNA. It did not affect the results if the samples were quickly thawed in a 37℃ water bath and detected immediately and the detect current velocity was lower than 300 cell/sec. The intra -CV was 7 28% and the internal CV was 8 92%. Mean and standard deviations of COMPαt were 8 7±11 0% in 511 healthy men. Because data present right skew distribution, the reference range of COMPαt is <26 8%. The results showed that COMPαt was positively correlated to the percentage of d sperm, MAD, percentage of sperm and negatively correlated to the survival rate and the percentage of a sperm. Conclusion Flow cytometric sperm chromatin structure assay can analyze sperm DNA injury rapidly, which deliver a new method to evaluate sperm sub clinical injury and male reproductivity and can foresee the successful rate of embryo transplantation.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2003年第10期616-619,共4页
Chinese Journal of Laboratory Medicine
