热和紫外线照射对丙型肝炎病毒JFH-1株灭活效果的研究  

Study on the effects of heat and ultraviolet light radiation on hepatitis C virus JFH-1 strain inactivation

作　　者：宋鸿硕[1] 石爽[1] 闫玲[1] 李奎[1] 庄辉[1] 

机构地区：[1]北京大学医学部病原生物学系,100191

出　　处：《中华微生物学和免疫学杂志》2009年第5期438-442,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家“十五”攻关课题（2001BA705B06）

摘　　要：目的应用HCV JFH-1株细胞培养系统，研究热和紫外线照射对HCV的灭活效果。方法感染性滴度为2．5×10^4 FFU／ml的丙型肝炎病毒JFH-1株（HCV JFH-1 strain）原液，经56℃水浴或紫外线照射不同时间后，感染Huh7-25-CD81细胞系，应用间接免疫荧光法测定病毒感染性滴度的动态变化。若被感染的细胞经盲传3代后，用间接免疫荧光法检测为阴性，则判定病毒原液已被完全灭活。结果感染性滴度为2．5×10^4 FFU／ml的HCV JFH-1株原液，经56℃水浴孵育10min、20min、30min后，其细胞的感染性滴度分别降至1．6×10^3 FFU／ml、3．1×10^2 FFU／ml和3．3×10 FFU／ml；该HCV JFH-1株原液暴露于紫外灯下（波长253．7nm，辐照强度≥60μW／cm^2）30cm处照射15s、30s、45s后，其细胞的感染性滴度分别降至1．0×10^3 FFU／ml、1．1×10^2 FFU／ml和2．7×10 FFU／ml。经56℃水浴孵育40min或紫外灯下照射（波长253．7nm，辐照强度≥60μW／cm^2）1min后，应用间接免疫荧光法检测为阴性，被感染的细胞经盲传3代后，间接免疫荧光法检测仍为阴性，证明该病毒液已被完全灭活。结论HCV JFH-1株对热和紫外线照射较为敏感，56℃下40min或紫外线照射（波长253．7nm，辐照强度≥60μW／cm^2，距离30cm）1min，可完全灭活HCV JFH-1株。Objective To investigate the inactivating effect of heat and ultraviolet （UV） light on HCV JFH-1 strain using the cell culture system. Methods The HCV JFH-1 virus stock, with an initial titer of 2.5×10^4 FFU/ml, was exposed in 56℃ water bath or to UV light for varying durations of time for exploring their inactivating effects on the virus. The kinetics of virus titer reduction was determined by an indirect immuno-fluorescence assay （ IFA ）. If the cells infected with the exposed virus stock were IFA negative after three blind passages, the virus stock was considered to be inactivated completely. Results After incubation of the HCV JFH-1 virus stock （2.5×10^4 FFU/ml）in 56℃ water bath for 10 min, 20 min and 30 min, the virus titers were reduced to 1.6×10^3 FFU/ml, 3.1 ×10^2 FFU/ml and 3.3×10 FFU/ml, respectively. The exposure of the virus stock to UV light （wavelength 253.7 nm, intensity ≥60 μW/cm/, 30 cm below the UV lamp） for 15 s, 30 s and 45 s resuhed in virus titer reduction to 1.0×10^3 FFU/ml, 1.1×10^2 FFU/ml and 2.7×10 FFU/ml, respectively. After 40 min incubation of the virus stock at 56℃, or 1 min exposure to UV light （wavelength 253.7 nm, intensity ≥60 μW/cm^2） the virus infectious titer was reduced below the detection limit of IFA, and the IFA was still negative even after three blind passages, indicating that the virus was inactivated completely. Conclusion HCV is sensitive to heat and UV light treatment. For HCV JFH-1 virus stock containing 2.5×10^4 FFU/ml virus, heat treatment at 56℃ for 40 min, or UV light exposure at an intensity of ≥60 μW/cm^2 for 1 min, resulting in complete virus inactivation.

关 键 词：丙型肝炎病毒 细胞培养 感染性滴度 灭活 

分 类 号：R51[医药卫生—内科学]