Abstract：
OBJECTIVE: To study the correlation, consistency, and variations between two assays of DNA fragmentation index based on acridine orange (AO) staining via AI-based fluorescence microscopy(AI-DFI), and flow cytometry (FCM-DFI) across multiple centers.
METHODS: We selected 421 male patients from Nanjing Drum Tower hospital ( Hospital G) (226 cases), Eastern Theatre General Hospital (Hospital J) (89 cases) and Jiangsu Province Hospital (Hospital S) (106 cases) . Semen samples from each patient were analyzed for routine semen parameters and for DFI using both AI fluorescence microscopy and flow cytometry. We studied the two methods\' stability as well as the correlation, consistency, and variation between the two methods\' results in various centers.
RESULTS: The two replicate studies\' results of AI-DFI and the three centers\' FCM-DFI for linear regression analysis indicated strong stability (R2>0.9).Overall(Group A), the AI-DFI results demonstrated good correlation and consistency with the FCM-DFI results of three centers (r>0.85；ICC>0.9).The semen specimens were categorized into two groups: normal specimen group (group B) and abnormal specimen group (group C) (including asthenozoospermia, oligospermia, and semen samples with high impurities).Group C\'s results showed a decline in correlation and consistency when compared to group A and group B, whereas group B\'s results showed a little rise in correlation and consistency when compared to group A. Although the consistency and correlation between the results of the two DFI testing methods in the three centers were good, there was still a significant difference between Groups A and C (P<0.05), and in Group B there was a significant difference between the two DFI testing methods only in Hospital G (p＝0.02), with no significant difference in Hospitals J and S (P> 0.05).
CONCLUSIONS: The two detection methods exhibit good stability and correlation. However, significant differences are observed in the DFI detection methods in samples with abnormal semen parameters and high complexity. The main reason for these significant differences may lie in the variations in detection principles. Each detection method has its own advantages, allowing clinical or research settings to choose between them based on laboratory conditions or specific requirements.
METHODS: We selected 421 male patients from Nanjing Drum Tower hospital ( Hospital G) (226 cases), Eastern Theatre General Hospital (Hospital J) (89 cases) and Jiangsu Province Hospital (Hospital S) (106 cases) . Semen samples from each patient were analyzed for routine semen parameters and for DFI using both AI fluorescence microscopy and flow cytometry. We studied the two methods\' stability as well as the correlation, consistency, and variation between the two methods\' results in various centers.
RESULTS: The two replicate studies\' results of AI-DFI and the three centers\' FCM-DFI for linear regression analysis indicated strong stability (R2>0.9).Overall(Group A), the AI-DFI results demonstrated good correlation and consistency with the FCM-DFI results of three centers (r>0.85；ICC>0.9).The semen specimens were categorized into two groups: normal specimen group (group B) and abnormal specimen group (group C) (including asthenozoospermia, oligospermia, and semen samples with high impurities).Group C\'s results showed a decline in correlation and consistency when compared to group A and group B, whereas group B\'s results showed a little rise in correlation and consistency when compared to group A. Although the consistency and correlation between the results of the two DFI testing methods in the three centers were good, there was still a significant difference between Groups A and C (P<0.05), and in Group B there was a significant difference between the two DFI testing methods only in Hospital G (p＝0.02), with no significant difference in Hospitals J and S (P> 0.05).
CONCLUSIONS: The two detection methods exhibit good stability and correlation. However, significant differences are observed in the DFI detection methods in samples with abnormal semen parameters and high complexity. The main reason for these significant differences may lie in the variations in detection principles. Each detection method has its own advantages, allowing clinical or research settings to choose between them based on laboratory conditions or specific requirements.
摘要：
目的：研究两者的相关性，一致性,以及通过基于AI的荧光显微镜（AI-DFI）基于吖啶橙（AO）染色的DNA片段化指数的两种测定之间的差异，和多个中心的流式细胞术（FCM-DFI）。
方法：我们选取南京鼓楼医院（G医院）的男性患者421例（226例），东剧院总医院（J医院）89例，江苏省人民医院（S医院）106例。使用AI荧光显微镜和流式细胞术分析每位患者的精液样品的常规精液参数和DFI。我们研究了这两种方法的稳定性以及相关性，一致性,两种方法之间的差异导致不同的中心。
结果：AI-DFI的两个重复研究结果和用于线性回归分析的三个中心\'FCM-DFI表明稳定性强（R2>0.9）。总体(A组),AI-DFI结果与三个中心的FCM-DFI结果具有良好的相关性和一致性（r>0.85；ICC>0.9）。将精液标本分为两组：正常标本组（B组）和异常标本组（C组）（包括弱精子症，少精子症,和高杂质的精液样品)。与A组和B组相比，C组的结果显示相关性和一致性下降，而B组的结果与A组相比，相关性和一致性略有上升。尽管三个中心的两种DFI测试方法的结果之间的一致性和相关性都很好，A组和C组之间仍有显著差异(P<0.05),B组两种DFI检测方法仅在G医院有显著性差异(p＝0.02),J、S医院差异无统计学意义(P>0.05)。
结论：两种检测方法具有良好的稳定性和相关性。然而,在精液参数异常和复杂性高的样品中，DFI检测方法存在显着差异。这些显著差异的主要原因可能在于检测原理的变化。每种检测方法都有自己的优点，允许临床或研究环境根据实验室条件或特定要求在它们之间进行选择。
方法：我们选取南京鼓楼医院（G医院）的男性患者421例（226例），东剧院总医院（J医院）89例，江苏省人民医院（S医院）106例。使用AI荧光显微镜和流式细胞术分析每位患者的精液样品的常规精液参数和DFI。我们研究了这两种方法的稳定性以及相关性，一致性,两种方法之间的差异导致不同的中心。
结果：AI-DFI的两个重复研究结果和用于线性回归分析的三个中心\'FCM-DFI表明稳定性强（R2>0.9）。总体(A组),AI-DFI结果与三个中心的FCM-DFI结果具有良好的相关性和一致性（r>0.85；ICC>0.9）。将精液标本分为两组：正常标本组（B组）和异常标本组（C组）（包括弱精子症，少精子症,和高杂质的精液样品)。与A组和B组相比，C组的结果显示相关性和一致性下降，而B组的结果与A组相比，相关性和一致性略有上升。尽管三个中心的两种DFI测试方法的结果之间的一致性和相关性都很好，A组和C组之间仍有显著差异(P<0.05),B组两种DFI检测方法仅在G医院有显著性差异(p＝0.02),J、S医院差异无统计学意义(P>0.05)。
结论：两种检测方法具有良好的稳定性和相关性。然而,在精液参数异常和复杂性高的样品中，DFI检测方法存在显着差异。这些显著差异的主要原因可能在于检测原理的变化。每种检测方法都有自己的优点，允许临床或研究环境根据实验室条件或特定要求在它们之间进行选择。
