表面增强拉曼光谱检测滤纸干血片用于新生儿先天性甲状腺功能减低症筛查的研究

The primary study on the newborn screening of congenital hypothyroidism with the dry blood spot in the filter paper by surface enhanced Raman spectroscopy

目的建立一种表面增强拉曼光谱（SERSp，也称拉曼分子指纹谱）分析技术检测滤纸干血片方法，探索用于筛查新生儿先天性甲状腺功能减低症（CH）的可能性和可靠性。方法本研究为病例对照研究。新生儿滤纸干血片标本和有关CH—hTSH筛查检测结果与诊断结论由作者在上海儿童医院新生儿筛查中心提供，SERSp实验在大连理工大学合作完成。选取经常规时间分辨荧光免疫法检测促甲状腺素（TSH）浓度筛查CH（CH—hTSH）后确诊阴性和阳性滤纸干血片标本各6份，检测滤纸干血片水溶液表面增强拉曼光谱，以725cm-1谱线为内标，分析得到的拉曼分子指纹谱；使用特征谱线分析，主成分分析方法（PCA）及PCA分群散点图分析，比较TSH检测结果。结果用725cm-1拉曼峰为内标，对照CH阴／阳性标本的SERSp实验结果，发现1373和1400cm-1两谱线用于分析CH的阴性和阳性的相对峰高差最大，提示1373和1400cm-1两条谱线可能为CH的特征谱线；而TSH的三条拉曼特征谱线（785、827和853cm-1）的阴性／阳性的相对峰高差异很小，用于TSH的特征峰进行CH-拉曼筛查尚存在困难；12例SERSp检测结果用主成分分析方法、PCA分群散点图和1373和1400cm-1两条CH特征谱分析方法均证实，CH-拉曼筛查与用荧光免疫检测CH-hTSH筛查的结果基本相符，其符合率为91．7％，CH-拉曼筛查方法的灵敏度为100％，特异性为83．3％。结论初步成功地开拓了一种采用SERSp检测分析的CH-拉曼筛查新方法，证实将足跟滤纸血片-SERSp分析技术应用于新生儿CH筛查有很好的前景。

Objective To develop a method of detecting dry blood spot （DBS） samples collected in the filter paper by using Surface Enhanced Raman Spectroscopy （ SERSp, also known as Raman molecular fingerprint spectrum） technology. The possibility and reliability of applying the technology to the newborn screening of congenital hypothyroidism （CH） was investigated. Methods The case-control study was used. The results of CH-hTSH screening test based on the DBS samples collected in the filter paper, and the clinical diagnosis were all provided by neonatal screening center of Shanghai Children＇s Hospital The results of SERSp analysis were provided by Dalian University of Technology. Six positive and six negative samples, which were confirmed respectively by CH-hTSH screening using time resolved fluoroimmunoassay （TRFIA） were chosen. With the spectrum lines of 725 cm- 1 as internal standard,the SERSp from the aqueous solution of these samples were analyzed. The specific spectrum line analysis, the principal component analysis （PCA） and the scatter diagram of PCA were used to compare the results of TSH test. Results With thespectrum lines of 725 cm as internal standard, the CH specific spectrum lines of 1373 cm-1 and 1400 -1 cm were discovered. They showed the most significant difference of the peak height between the positive and negative samples of CH, while TSH specific spectrum lines of 785 cm-1, 827 cm- l and 853 cm- 1 failed to distinguish positive from negative samples with distinct peak height. The consistency between the results of CH-Raman Screening and CH-hTSH Screening was 91.7% （ 11/12）, which was proven by PCA, the scatter diagram of PCA and the specific spectrum line analysis. The sensitivity of CH-Raman Screening was 100% and the specificity was 83.3%. Conclusion There is a new CH-Raman Screening method using the SERSp analysis, which was proven to be a promising technology in the newborn screening of congenital hypothyroidism with the DBS in the filter paper.