基于拉曼光谱技术的鼻咽癌无损诊断研究

发布时间：2018-04-19 16:22

本文选题：鼻咽癌 + RS　；参考：《福建医科大学》2016年博士论文

【摘要】：拉曼光谱技术(Raman Spectroscopy,RS)及表面增强拉曼光谱技术(Surface Enhanced Raman Spectroscopy,SERS)是当今国际生物医学领域的研究热点,本研究结合临床上鼻咽癌诊断的需要,利用RS和SERS技术的高灵敏性,检测鼻咽癌患者组织,血液,唾液等样本,并结合PCA(Principle Components Analysis,主成分分析)及LDA(linear discriminant analysis,线性判别分析)多元统计模型,试图研发一种快速、无损、客观、准确的基于RS及SERS技术的鼻咽癌早期诊断方法。主要研究工作包括五部分:1.常规拉曼光谱技术(RS)在早晚期鼻咽癌组织中诊断价值研究目的:本研究旨在比较鼻咽癌患者早晚期病理组织的拉曼光谱差异,评估拉曼光谱技术在鼻咽癌早晚期诊断中的价值。材料与方法:使用常规拉曼光谱技术(RS)分别检测30例鼻咽癌早期(T1-2)患者与46例鼻咽癌晚期(T3-4)患者组织标本的拉曼光谱,利用主成分分析法(PCA)并结合线性判别分析法(LDA)建立常规拉曼光谱技术诊断方法的多元统计算法模型。结果:鼻咽癌早期与晚期患者组织RS信号差异明显,PCA-LDA判别区分的灵敏性、特异性和准确性分别为70%、78.3%及75%。结论:RS技术结合PCA-LDA统计分析能够较好地区分鼻咽癌患者早晚期组织标本,表明在早晚期鼻咽癌判别上RS方法可行。2.鼻咽癌患者唾液的表面增强拉曼光谱技术(SERS)直接法检测研究目的:本文目的在于预测基于受试者唾液的表面增强拉曼光谱技术(SERS)对鼻咽癌的无损诊断意义。材料与方法:共收集52例鼻咽癌患者及60例健康志愿者的唾液样本,分别检测其SERS光谱,并对获得的SERS光谱进行谱峰归属分析。结果:鼻咽癌患者与健康志愿者的唾液SERS光谱差异明显,主要位于447,496,635,729,1134,1270,和1448 cm-1处,谱峰归属分析提示,健康受试者与鼻咽癌患者之间唾液中糖类、蛋白、核酸及脂肪酸存在差别。统计结果分析显示,基于鼻咽癌唾液SERS检测技术诊断鼻咽癌的特异度为91.7%,灵敏度为82.7%,准确率为87.5%。结论:本研究表明,直接法检测唾液表面增强拉曼光谱技术(SERS)结合PCA-LDA分析可以较好地区分鼻咽癌患者与健康志愿者,该技术对于鼻咽癌无损诊断具有潜在的临床应用价值。3.不同T分期鼻咽癌血浆表面增强拉曼光谱(SERS)检测研究目的:本研究评估基于血浆无标记的表面增强拉曼光谱(SERS)技术区分不同T分期鼻咽癌血浆的可行性。Au(金)纳米颗粒作为SERS活性的纳米结构直接与人类血液混合成等离子体获得增强的拉曼散射信号。材料与方法:我们共检测60例健康志愿者血浆样本,25例T1期鼻咽癌和75例T2-T4鼻咽癌患者血浆样本。结果:表明T1期与健康受试者,T2-T4期与健康受试者的血浆,其差异诊断敏感性分别为84%和92%,特异性为83.3%和95%,准确性为83.5%和93.3%。然而,T1期与T2-T4鼻咽癌血浆之间差异的敏感性、特异性和准确性仅有64%,62.7%和63%,区分度不够理想;这个探索性研究表明基于SERS纳米生物技术结合PCA-LDA统计方法可以实现早期T1和健康志愿者;T2-T4和健康志愿者的血浆之间的区别,但早晚期之间差异区分较难。结论:该方法可以作为临床上诊断鼻咽癌重要的辅助手段,但要区分早晚期鼻咽癌还有较大的挑战。4.鼻咽癌组织DNA的表面增强拉曼光谱技术(SERS)检测研究目的:研究表面增强拉曼光谱技术在鼻咽癌早晚期及健康受试者鼻咽组织DNA的差异,探讨表面增强拉曼光谱技术在鼻咽癌组织DNA分子诊断中的应用价值。材料与方法:本研究采用SERS技术检测鼻咽癌病理组织与正常鼻咽组织中DNA的差别,分别提取26例早期鼻咽癌、35例晚期鼻咽癌患者病理组织与30例健康受试者鼻咽组织的DNA进行SERS检测,对比分析三组拉曼散射光谱的差异。结果:研究发现鼻咽癌早、晚期组织与健康受试者鼻咽组织的DNA在谱峰上存在着明显差异,经过PCA-DFA分析发现,早期鼻咽癌组织诊断的灵敏度为80.8%,特异度为84.6%,准确率为86.8%,晚期鼻咽癌组织诊断的灵敏度为85.7%,特异度为82.1%,准确率为92.3%,具有较高的诊断准确率。结论:本研究提示基于SERS纳米生物技术结合PCA-DFA统计分析作为临床早晚期鼻咽癌的补充检测具有较好的前景。5.鼻咽内窥镜下活体鼻咽癌荧光成像和拉曼光谱检测目的:鼻咽癌最常用的筛选方法是鼻咽镜下对可疑病变部位进行组织病理活检,但可能造成漏诊和误诊。为避免漏诊及误诊,我们探讨鼻咽纤维镜下鼻咽癌体内拉曼光谱检测的可行性。材料与方法:本文采用鼻咽内窥镜下在体鼻咽癌荧光成像及拉曼光谱技术,检测了鼻咽癌活体荧光图像和拉曼光谱。结果:首次获得了鼻咽癌实时活体拉曼光谱。结论:表明将鼻咽镜下活体荧光成像和拉曼光谱技术相结合,提高鼻咽癌诊断的有效性和客观性方面具有一定的潜力。
[Abstract]:Raman Spectroscopy (RS) and surface enhanced Raman spectroscopy (Surface Enhanced Raman Spectroscopy, SERS) are the hot spots in the field of international biomedical research. This study combines the high sensitivity of RS and SERS techniques to detect the tissues, blood, saliva and other samples of nasopharyngeal carcinoma patients with the needs of the diagnosis of the clinical nasopharyngeal carcinoma. This paper, combined with PCA (Principle Components Analysis, principal component analysis) and LDA (linear discriminant analysis, linear discriminant analysis) multivariate statistical model, attempts to develop a fast, nondestructive, objective and accurate method for early diagnosis of nasopharyngeal carcinoma based on RS and SERS technology. The main research work includes five parts: 1. conventional Raman spectroscopy (R) S) a study on the diagnostic value of nasopharyngeal carcinoma in the early and late stage of nasopharyngeal carcinoma. The purpose of this study was to compare the differences in the Raman spectrum of the early and late pathological tissues of nasopharyngeal carcinoma and to evaluate the value of Raman spectroscopy in the early diagnosis of nasopharyngeal carcinoma. Materials and methods: 30 patients with nasopharyngeal carcinoma (T1-2) and 46 were detected by conventional Raman spectroscopy (RS). The Raman spectrum of the tissue specimens of patients with advanced nasopharyngeal carcinoma (T3-4), using the principal component analysis (PCA) and linear discriminant analysis (LDA) to establish the multivariate statistical algorithm model of the conventional Raman spectroscopy diagnostic method. Results: the difference of RS signal in the early and late stage of nasopharyngeal carcinoma is obvious, and the sensitivity and specificity of PCA-LDA discriminant distinction And the accuracy is 70%, 78.3% and 75%. conclusion: RS technique combined with PCA-LDA statistical analysis can better distinguish the early and late tissue specimens of nasopharyngeal carcinoma patients, indicating that the RS method is feasible for the direct detection of saliva of.2. nasopharyngeal carcinoma patients in the early and late stage of nasopharyngeal carcinoma identification. The purpose of this article is to preview the purpose of this study. A total of 52 cases of nasopharyngeal carcinoma and 60 healthy volunteers were collected from the saliva samples of 52 cases of nasopharyngeal carcinoma and 60 healthy volunteers. The spectra of the saliva of 52 cases of nasopharyngeal carcinoma and 60 healthy volunteers were collected, and the spectrum peaks of the obtained SERS spectra were analyzed. The difference of SERS spectra in saliva was obvious, mainly located at 44749663572911341270, and 1448 cm-1. The analysis of spectrum peak attribution indicated that there were differences in saliva sugar, protein, nucleic acid and fatty acids between healthy subjects and nasopharyngeal carcinoma patients. The analysis showed that the specificity of the diagnosis of nasopharyngeal carcinoma based on the SERS detection technique of nasopharyngeal carcinoma was 91.. 7%, the sensitivity is 82.7% and the accuracy is 87.5%. conclusion: This study shows that direct detection of saliva surface enhanced Raman spectroscopy (SERS) combined with PCA-LDA analysis can be a better region for nasopharyngeal carcinoma and healthy volunteers. This technique has potential clinical application value for nasopharyngeal carcinoma nondestructive diagnosis,.3. different T staging nasopharyngeal carcinoma plasma surface The purpose of enhanced Raman spectroscopy (SERS) detection: This study evaluated the feasibility of using plasma unlabeled surface enhanced Raman spectroscopy (SERS) to distinguish the feasibility of different T staging nasopharyngeal carcinoma plasma,.Au nanoparticles were used as SERS active nanostructures to obtain enhanced Raman scattering signals directly from human blood mixed plasma. And methods: We measured plasma samples of 60 healthy volunteers, 25 cases of T1 nasopharyngeal carcinoma and 75 patients with T2-T4 nasopharyngeal carcinoma. Results: the diagnostic sensitivity of phase T1 and healthy subjects, T2-T4 and healthy subjects was 84% and 92%, the specificity was 83.3% and 95%, the accuracy was 83.5% and 93.3%., however, T1 phase and 93.3%. The sensitivity, specificity and accuracy of plasma differences between T2-T4 nasopharyngeal carcinoma (nasopharyngeal carcinoma), specificity and accuracy were only 64%, 62.7% and 63%, and the difference was not ideal; this exploratory study showed that SERS based nanobiotechnology combined with PCA-LDA statistics could achieve early T1 and healthy volunteers, and the difference between T2-T4 and healthy volunteers, but the difference between early and late stages Conclusion: this method can be used as an important auxiliary method in clinical diagnosis of nasopharyngeal carcinoma, but it is necessary to distinguish the early and late stage nasopharyngeal carcinoma and the large challenge of the.4. nasopharyngeal carcinoma tissue DNA surface enhanced Raman spectroscopy (SERS) detection research purpose: the study of surface enhanced Raman spectroscopy in nasopharyngeal carcinoma early and healthy subjects nose The difference between the DNA of the pharynx tissue and the application value of surface enhanced Raman spectroscopy in the diagnosis of DNA in nasopharyngeal carcinoma. Materials and methods: This study used SERS to detect the difference between the pathological tissue of nasopharyngeal carcinoma and the normal nasopharyngeal tissue, and to extract 26 cases of early nasopharyngeal carcinoma, 35 cases of advanced nasopharyngeal carcinoma and 30 cases of health. The DNA of the nasopharynx tissues of the subjects was detected by SERS. The differences between the three groups were compared and analyzed. Results: the study found that the nasopharyngeal carcinoma was early, and the DNA of the nasopharynx tissues in the late tissues and the healthy subjects were significantly different. The sensitivity of the early nasopharyngeal carcinoma was 80.8% and the specificity was 84.6% after PCA-DFA analysis. The accuracy rate is 86.8%, the sensitivity of the advanced nasopharyngeal carcinoma tissue is 85.7%, the specificity is 82.1%, the accuracy is 92.3%, and the diagnostic accuracy is high. Conclusion: This study suggests that the SERS nano biotechnology combined with the statistical analysis of PCA-DFA is a good prospect for nasopharyngeal carcinoma in the early and late stage of the nasopharyngeal carcinoma with.5. nasopharynx endoscopy. Fluorescence imaging and Raman detection of somatic nasopharyngeal carcinoma: the most commonly used method of screening nasopharyngeal carcinoma is the histopathological biopsy of the suspected site under the nasopharynx, but it may cause missed diagnosis and misdiagnosis. In order to avoid missed diagnosis and misdiagnosis, we discuss the feasibility of Raman spectrum detection in nasopharyngeal carcinoma in nasopharyngeal carcinoma. Materials and methods: The fluorescence and Raman spectra of nasopharyngeal carcinoma were detected by fluorescence imaging and Raman spectroscopy under nasopharyngeal endoscopy. Results: the real time living body Raman spectra of nasopharyngeal carcinoma were obtained for the first time. Conclusion: it is shown that the combination of the living fluorescent imaging and Raman spectroscopy under nasopharynx can improve the effectiveness of the diagnosis of nasopharyngeal carcinoma. There is a certain potential in objectivity.

【学位授予单位】：福建医科大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R739.63

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