基于精浆SERS光谱技术的男性无精子症诊断方法研究

Azoospermia is the most serious case in male infertility, limitations of clinical diagnosis are time-consuming, cumbersome operation as well as potential risk of unpredicted damage to other organs during a testicular biopsy. Seminal plasma serves not only as the carrier to sperm, but also the "soil" and energy source for spermatozoa. Seminal plasma is rich in biochemical components and the corresponding changes can directly imply the early pathological state, which can be considered as a most important entry for detection. Based on our previous work, this project develops a new method that employs capillary tube SERS substrate to interrogate seminal plasma samples from patients suffering varying degrees of azoospermia. Through band assignment and extraction of SERS characteristic peaks, qualitative analysis of biochemical substances in seminal plasma was conducted. In addition, multivariate statistical analysis such as partial least squares (PLS) was carried out to quantitatively determine the levels of vital components (such as amino acids, glucosidase, and fructose) in seminal plasma. With the combination of qualitative and quantitative results, SERS spectroscopy based diagnostic model for diagnosis and differentiation of male azoospermia was built. Therefore, the project is expected to address the current shortcomings in clinical diagnosis of azoospermia such as high cost, time-consuming and potentially testicular biopsy damage, thus providing a rapid, objective and non-destructive method for differential diagnosis and classifications of azoospermia in the future to assist the objective diagnosis of male infertility, which will definitely have great significance.

无精子症是男性不育的最严重情形，其临床诊断存在耗时、操作繁琐、睾丸活检有损等问题。精浆不仅是精子的载体，更是其能量来源和生存“土壤”；精浆生化成分丰富，相关的生化指标改变可直接、灵敏的反映出男性生殖器官疾病的早期病理状态，也是生化检测最重要的切入点。本项目在前期工作的基础上设计新型毛细管SERS检测基底，获得无精子症患者不同病理的精浆SERS光谱数据；并通过谱峰归属、特征谱峰提取，定性分析比较精浆中生化物质的差异；此外，结合以偏最小二乘法（PLS）为主的多变量统计分析方法，开展精浆中重要生化成分（氨基酸、葡萄糖苷酶和果糖等）含量的定量检测；综合定性定量数据，构建检测、诊断区分无精子症的拉曼光谱诊断模型。因此，该项目研究有望针对目前男性无精子症诊断高成本、耗时、潜在睾丸活检损伤的不足，提供无精子症诊断和类型区分的快速、客观和无损新方法，进而有助于协助男性不育的客观诊断，具有重要的意义。