摘要
目的研究亚甲蓝光化学法(methylene blue photochemistry,MB-P)降解病毒核酸的作用机制,为该方法在临床血液制品病毒灭活中的应用提供理论依据。方法以丙型肝炎病毒(HCV)为研究对象,选择HCV基因组中相对保守的5′-NCR为模板,应用荧光定量PCR技术观察不同处理时间HCV RNA降解情况。同时,对体外转录的同区段的HCV RNA在无蛋白成分的代血浆(羟乙基淀粉20-氯化钠注射液)及PBS缓冲液中的降解模式进行分析。结果MB-P处理条件下,HCV RNA含量呈对数形式下降,该方法对体外转录RNA的灭活效率低于对血浆中病毒颗粒的灭活效率。结论亚甲蓝能作用于病毒核酸,在光照条件下使RNA迅速降解,从而达到病毒灭活的效果。
Objective In order to research on the mechanisms of the decrease of hepatitis C viral （HCV） RNA with methylene blue photochemistry （MB-P） treatment, and to provide theoretic bases for clinic applications on viral inactivation of blood products. Methods The conservative 5′-NCR of HCV genome RNA was selected as the research template. After being exposed in the light for different periods with methylene blue ,HCV RNA in the plasma was extracted and analyzed quantitatively with fluorescence quantitation PCR. Meanwhile, the same experiment was performed on in vitro transcripted 5′-NCR of HCV RNA, which was resolved in plasma substitute （hydroxyethyl starch 20-sodium chloride solution） and PBS buffer without any component of protein. Results The quantity of HCV RNA decreased in a logarithmic way with MB-P treatment, and the efficiency of inactivation on in vitro transcripted HCV RNA was lower than that of HCV virus in plasma. Conclusion With light exposure,methylene blue could act on viral nucleic acid and break RNA chain to decrease the quantity of viral RNA, whereafter inactivate the virus finally.
出处
《临床输血与检验》
CAS
2007年第3期193-197,共5页
Journal of Clinical Transfusion and Laboratory Medicine
基金
上海市医学重点学科建设项目(No.05III003)
上海市卫生局科技发展基金项目(No.044037)资助
关键词
病毒灭活
亚甲蓝光化学法
荧光定量PCR
Viral inactivation Methylene blue photochemistry Fluorescence quantitation PCR
