应用拉曼光谱技术优选人类发育潜力胚胎研究  

Application of Raman spectroscopy in selecting potential cleavage embyos

作　　者：徐维海[1] 张岭 张琳 李施施 舒静[1] XU Wei-hai;ZHANG Ling;ZHANG Lin;LI Shi-shi;SHU Jing(Zhejiang Provincial People＇s Hospital,Hangzhou,Zhejiang 310014,China)

机构地区：[1]浙江省人民医院生殖内分泌科,浙江杭州310014 [2]杭州医学院附属人民医院

出　　处：《预防医学》2018年第2期144-147,152,共5页CHINA PREVENTIVE MEDICINE JOURNAL

基　　金：浙江省医药卫生科技计划项目(2017KY219)

摘　　要：目的建立应用拉曼光谱技术优选具有发育潜力胚胎的实验方法。方法选取100份用于囊胚培养的第3天卵裂期胚胎培养液(D3培养液),确定合适的测定时间,获得原始拉曼光谱,通过滤波、基线校正和光谱归一化预处理,获得真实拉曼光谱和用于分析比较的光谱模型。测定173份D3培养液拉曼光谱曲线,评估其优选发育潜力胚胎的应用价值。结果采用≥240 s的光谱仪积分时间可获得分辨度较好的原始拉曼光谱,预处理后获得的真实拉曼光谱的特征性位移为942、1 346、1 456和1 656 cm^(-1)。阳性曲线和阴性曲线模型的光谱差异位于1 100~1 800 cm^(-1)。不同位移的真实拉曼光谱强度比较,差异均无统计学意义(P>0.05)。173份D3培养液获得阳性曲线80份,阴性曲线70份,中性曲线23份,囊胚形成率依次为85.00%、22.86%和65.22%,差异有统计学意义(P<0.001)。拉曼光谱曲线预测囊胚形成效率的灵敏度为83.8%,特异度为73.0%,Kappa值为0.572。结论通过改良光谱收集条件,并对原始拉曼光谱进行预处理,可获得清晰、真实、可比性好的拉曼光谱模型,采用本方法筛选的潜力胚胎与囊胚形成呈中度一致。Objective To develop a method with Raman spectroscopy for selecting embryos with high developing potential.Methods A total of 100 samples of D3 spent cultural medium for embryos assigned to blastocyst culture were collected,and theRaman spectroscopy produced with optimal integration time. The original spectrums retrieved were processed by filtration,baseline correction and normalization methods to form characteristic spectral for the spent medium of cleavage embryo,suitablefor analysis. A further 173 samples were collected to analyze the clinical relevance of Raman spectroscopy in selecting thepotential embryos. Results Original Raman spectra with good resolution can be obtained with an integral time over 240 seconds.The characteristic spectral shift of embryo culture medium was 942,1 346,1 456 and1 656 cm^-1after processing. The spectralshift between the positive curve and the negative model was 1 100-1 800 cm^-1. There was no significant difference in the intensityof real Raman spectral with different shifts（P〉0.05 ） . A total of 80 positive curves,70 negative curves,23 neutral curves wereobtained from the 173 D3 samples,and the blastocyst formation rates were 85.00%,22.86% and 65.22%,respectively,withstatistically significant difference（P〈0.001） . The sensitivity,specificity and Kappa value for the Raman spectrum curvepredicting the blastocyst formation were 83.8%,73.0% and 0.572,respectively. Conclusion A clear,real and comparableRaman spectrum model could be obtained by improving the spectral collection conditions and preprocessing the original Ramanspectrum. The potential embryo screened by this method is moderately consistent with that of blastocyst.

关 键 词：体外受精 胚胎 拉曼光谱 发育 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]