运用FQ-PCR技术监测亚甲蓝光化学法对血浆丙型肝炎病毒的灭活效果  

Effect of Methylene Blue and Visible Light Monitored by Real-time PCR on the Inactivation of Plasma Hepatitis C Virus.

作　　者：郑岚[1] 黄宇闻[1] 莫琴[1] 王迅[1] 许莉萍[1] 陆萍[1] 凌冰[1] 钱开诚[1] 曹文俊[2] 

机构地区：[1]上海市血液中心,200051 [2]上海第二医科大学附属瑞金医院,200025

出　　处：《诊断学理论与实践》2002年第2期102-104,共3页Journal of Diagnostics Concepts & Practice

摘　　要：目的:通过荧光定量PCR（FQ-PCR）技术对血浆病毒灭活前后丙型肝炎病毒RNA（HCV-RNA）浓度的测定,判断亚甲蓝（MB）光化学法灭活血浆丙型肝炎病毒的效果,为临床判别病毒灭活效果提供直接和客观依据。方法:4份经证实为HCV-RNA阳性的血浆分别加入MB,终浓度为1μmol/L,经可见光（大于30000Lux）照射不同时间（0、5、10、15和30min）后,分别用FQ-PCR测定这些血浆的HCV-RNA浓度,并与未用MB处理的血浆作比较,判断随照射时间不同,HCV被灭活的效果。结果:4份HCV-RNA阳性病人的血浆未经处理时其病毒核酸浓度分别为8.0X105拷贝/ml、1.7X106拷贝/ml、1.6X106拷贝/ml和 3.1X105拷贝/ml,加入MB未经照射时HCV-RNA浓度即明显下降,可见光照射5 min后HCV-RNA浓度分别下降为未经任何处理时浓度的18.75％、23.36％、4.66％和1.27％,随照射时间延长,病毒核酸浓度逐渐下降,照射30min后,HCV-RNA浓度均下降为零。结论:用MB加光照30min处理,可达到将血浆丙肝病毒灭活的效果;FQ-PCR可定量检测MB光化学法灭活丙肝病毒的过程。Objective: To evaluate the effect of methylene blue and visible light by the method of real-time PCR on the in-activation of plasma hepatitis C virus, and to establish a direct and impersonal basis for the surveillance of viral inac-tivation. Methods; Four samples of plasma of HCV-RNA positive were added with methylene blue up to the final concentration of 1 μmol/L. Then the plasma was processed with visible light radiation)30000 Lux) for 0, 5, 10, 15 and 30 min respectively. Moreover, the concentration of HCV-RNA samples was quantitatively detected by the method of real-time PCR and compared with untreated samples to evaluate the effect of HCV inactivation and its relationship with the time of radiation. Results: The original concentration of HCV-RNA was 8.0x105, 1.7X 106, 1.6x106 and 3.1x105 copies/ml respectively. After being added with methylene blue, the concentration of HCV-RNA was significantly decreased. And after 5 min of radiation, the concentration decreased to 18.75%, 23.36%, 4.66% and 1.27% in the proportion of the original concentration, after 30 min of radiation, the concentration was completely inactivated. Conclusions: The plasma HCV could be inactivated by methylene blue and visible light radiation for 30 min. Real-time PCR could be used to quantitatively monitor the whole process of inactivation of plasma Hepatitis C virus with methylene blue photochemistry.

关 键 词：FQ-PCR技术 亚甲蓝光化学法 血浆 丙型肝炎病毒 病毒灭活 

分 类 号：R512.63[医药卫生—内科学]