{"title":"鼻咽癌患者外周血eb病毒DNA的光电化学测定","authors":"K. Kusmardi, A. Tedjo, Y. H. Midoen, M. Adrian","doi":"10.1063/1.5139371","DOIUrl":null,"url":null,"abstract":"Epstein-Barr Virus (EBV) DNA presents in the serum of nasopharyngeal carcinoma (NPC) patients indicates the existence of the viral DNA in the circulation and may be used as an early marker in NPC diagnosis. EBV DNA isolated from the serum and then amplified by using polymerase chain reaction (PCR) could be used to assess the response of the DNA to various NPC therapies. This study compares the results of EBV DNA measurement in the serum of NPC patients using photoelectrochemical technique and PCR. The presence of a specific oligonucleotide sequence of a standard DNA (EBNA-1) may be detected by photoelectrochemical hybridization of the standard with a DNA probe immobilized on TiO2 electrode, in which the DNA is intercalated by an electrochromic material. The purpose of this study is to obtain a photoelectrochemical system with a relatively good response and sensitivity to the changes in the concentration of EBV DNA in the peripheral blood serum of NPC patients. This research attempts to provide a device and methods that are simple, relatively inexpensive and easy to use, yet sensitive enough to see the prognosis of NPC patients. This study also attempts to develop the measurement instrumentation technology in the field of biomedical research. The photoelectrochemical system of this research used anthocyanin as DNA intercalator. The anthocyanin was isolated from purple cabbage leaves (also known as red cabbage), strawberries, and grapes and the extractant of these anthocyanins were ethanol and water. The best extraction with the highest level of total anthocyanin was achieved by alcohol as extractant. Alcohol gave higher yield of anthocyanin compared to water and ethyl acetate. Purple cabbage leaves have the highest level of anthocyanin compared to strawberry fruits and grapes. Anthocyanins contained in the extract of purple cabbage, strawberries, and grapes may function as DNA intercalators. Electrochromic substance used as DNA intercalator produces difference between the current strength generated by photoelectrochemical system with light and without light. There is a very strong correlation between diluted concentration of DNA and current strength shown by the electrochemical system of this study. There is also correlation between EBV EBNA-1 DNA concentration readings by using photoelectrochemical system and PCR. The sensitivity and specificity to EBV DNA in the serum of NPC patients of photoelectrochemical techniques are significantly higher compared to PCR.Epstein-Barr Virus (EBV) DNA presents in the serum of nasopharyngeal carcinoma (NPC) patients indicates the existence of the viral DNA in the circulation and may be used as an early marker in NPC diagnosis. EBV DNA isolated from the serum and then amplified by using polymerase chain reaction (PCR) could be used to assess the response of the DNA to various NPC therapies. This study compares the results of EBV DNA measurement in the serum of NPC patients using photoelectrochemical technique and PCR. The presence of a specific oligonucleotide sequence of a standard DNA (EBNA-1) may be detected by photoelectrochemical hybridization of the standard with a DNA probe immobilized on TiO2 electrode, in which the DNA is intercalated by an electrochromic material. 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引用次数: 0

摘要

鼻咽癌(NPC)患者血清中存在eb病毒(EBV) DNA,提示该病毒DNA在血液循环中存在,可作为鼻咽癌诊断的早期标志物。从血清中分离EBV DNA,用聚合酶链反应(PCR)扩增EBV DNA,可用于评价EBV DNA对不同鼻咽癌治疗的反应。本研究比较了光电化学技术和PCR技术在鼻咽癌患者血清中检测EBV DNA的结果。用固定在TiO2电极上的DNA探针,通过电致变色材料嵌入DNA,对标准DNA (EBNA-1)进行光电杂交,可以检测到标准DNA (EBNA-1)中特定寡核苷酸序列的存在。本研究的目的是获得一种对鼻咽癌患者外周血EBV DNA浓度变化具有较好响应和敏感性的光电化学系统。本研究试图提供一种简单,相对便宜,易于使用,但足够敏感的仪器和方法来观察鼻咽癌患者的预后。本研究也尝试在生物医学研究领域发展测量仪器技术。本研究的光电化学体系以花青素为DNA插层剂。花青素是从紫甘蓝叶(也称为红甘蓝)、草莓和葡萄中分离出来的,提取剂为乙醇和水。以醇为萃取剂提取花青素效果最佳,总花青素含量最高。与水和乙酸乙酯相比,醇的花青素得率更高。紫甘蓝的叶子中花青素的含量高于草莓和葡萄。紫甘蓝、草莓和葡萄提取物中含有的花青素可能具有DNA插入剂的功能。电致变色物质作为DNA插层剂,在有光和无光的情况下,光电化学系统产生的电流强度存在差异。本研究电化学系统显示的DNA稀释浓度与电流强度之间存在很强的相关性。光电化学与PCR检测的EBNA-1 DNA浓度也存在相关性。光电化学技术对鼻咽癌患者血清EBV DNA的敏感性和特异性均明显高于PCR。鼻咽癌(NPC)患者血清中存在eb病毒(EBV) DNA,提示该病毒DNA在血液循环中存在,可作为鼻咽癌诊断的早期标志物。从血清中分离EBV DNA,用聚合酶链反应(PCR)扩增EBV DNA,可用于评价EBV DNA对不同鼻咽癌治疗的反应。本研究比较了光电化学技术和PCR技术在鼻咽癌患者血清中检测EBV DNA的结果。用固定在TiO2电极上的DNA探针,通过电致变色材料嵌入DNA,对标准DNA (EBNA-1)进行光电杂交,可以检测到标准DNA (EBNA-1)中特定寡核苷酸序列的存在。本研究的目的是获得一种对鼻咽癌患者外周血EBV DNA浓度变化具有较好响应和敏感性的光电化学系统。本研究试图提供一种设备…
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Photoelectrochemical measurement of Epstein-Barr virus DNA in peripheral blood serum of nasopharyngeal carcinoma patients
Epstein-Barr Virus (EBV) DNA presents in the serum of nasopharyngeal carcinoma (NPC) patients indicates the existence of the viral DNA in the circulation and may be used as an early marker in NPC diagnosis. EBV DNA isolated from the serum and then amplified by using polymerase chain reaction (PCR) could be used to assess the response of the DNA to various NPC therapies. This study compares the results of EBV DNA measurement in the serum of NPC patients using photoelectrochemical technique and PCR. The presence of a specific oligonucleotide sequence of a standard DNA (EBNA-1) may be detected by photoelectrochemical hybridization of the standard with a DNA probe immobilized on TiO2 electrode, in which the DNA is intercalated by an electrochromic material. The purpose of this study is to obtain a photoelectrochemical system with a relatively good response and sensitivity to the changes in the concentration of EBV DNA in the peripheral blood serum of NPC patients. This research attempts to provide a device and methods that are simple, relatively inexpensive and easy to use, yet sensitive enough to see the prognosis of NPC patients. This study also attempts to develop the measurement instrumentation technology in the field of biomedical research. The photoelectrochemical system of this research used anthocyanin as DNA intercalator. The anthocyanin was isolated from purple cabbage leaves (also known as red cabbage), strawberries, and grapes and the extractant of these anthocyanins were ethanol and water. The best extraction with the highest level of total anthocyanin was achieved by alcohol as extractant. Alcohol gave higher yield of anthocyanin compared to water and ethyl acetate. Purple cabbage leaves have the highest level of anthocyanin compared to strawberry fruits and grapes. Anthocyanins contained in the extract of purple cabbage, strawberries, and grapes may function as DNA intercalators. Electrochromic substance used as DNA intercalator produces difference between the current strength generated by photoelectrochemical system with light and without light. There is a very strong correlation between diluted concentration of DNA and current strength shown by the electrochemical system of this study. There is also correlation between EBV EBNA-1 DNA concentration readings by using photoelectrochemical system and PCR. The sensitivity and specificity to EBV DNA in the serum of NPC patients of photoelectrochemical techniques are significantly higher compared to PCR.Epstein-Barr Virus (EBV) DNA presents in the serum of nasopharyngeal carcinoma (NPC) patients indicates the existence of the viral DNA in the circulation and may be used as an early marker in NPC diagnosis. EBV DNA isolated from the serum and then amplified by using polymerase chain reaction (PCR) could be used to assess the response of the DNA to various NPC therapies. This study compares the results of EBV DNA measurement in the serum of NPC patients using photoelectrochemical technique and PCR. The presence of a specific oligonucleotide sequence of a standard DNA (EBNA-1) may be detected by photoelectrochemical hybridization of the standard with a DNA probe immobilized on TiO2 electrode, in which the DNA is intercalated by an electrochromic material. The purpose of this study is to obtain a photoelectrochemical system with a relatively good response and sensitivity to the changes in the concentration of EBV DNA in the peripheral blood serum of NPC patients. This research attempts to provide a device an...
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