摘要
目的 探索温度对血清中乙型肝炎病毒（HBV）感染力的影响。 方法 将含有高浓度HBV DNA（1.33×108拷贝/ml）的HBV阳性血清分装于1.5 ml无菌离心管内，每管100 μl共23管，密封后各有5管置37、56、65 ℃恒温水浴箱内孵育，分别于0、30、60、120、600 min后各取出1管；各有4管放置在98 ℃恒温水浴箱孵育和沸水锅中，放置0、5、10、30 min后各取1管，进行HepG-2细胞感染试验，共孵育18 h后洗弃接种液加新鲜培养基，48 h后收集细胞，实时荧光定量PCR（FQ-PCR）检测HBV DNA，以0 min处理组为对照组。结果 HBV阳性血清在低热处理30、60、120、600 min后感染HepG-2细胞，细胞内HBV DNA拷贝数在37 ℃时分别是（4.85±1.71）×105、（3.85±1.76）×105、（1.67±0.67）×105、（7.86±1.03）×104拷贝/ml，56 ℃时分别是（4.01±0.16）×105、（9.77±0.97）×104、（6.36±0.65）×104、（5.05±0.24）×103拷贝/ml；65 ℃时60、120 min处理组感染细胞内HBV DNA拷贝数分别是（5.15±7.28）×103、（7.56±10.6）×102拷贝/ml，是对照组[（6.79±1.48）×105拷贝/ml]的10-2、10-3倍，不同温度处理HBV阳性血清后感染HepG-2细胞、细胞内HBV DNA的含量差异有统计学意义（F处理温度=104.4，P＜0.001），不同处理时间后的感染细胞内HBV DNA的含量差异有统计学意义（F处理时间=144.0，P＜0.001），温度和处理时间有交互作用（F处理时间＊处理温度=23.6，P＜0.001），高温处理时，98 ℃孵育10 min、煮沸5 min处理时感染细胞内HBV DNA拷贝数分别为（3.02±4.26）×102、（4.31±6.09）×102，比对照组[（6.79±1.48）×105拷贝/ml]下降10-3倍，98 ℃孵育30 min、煮沸10 min后感染细胞内6次检测均未检出HBV DNA。结论 离体血清HBV感染力在低热条件下相对稳定，相对高温条件下短时间即可使其感染力降低。
Objective This article was to explore the impact of temperature on hepatitis B virus infectivity. Methods HBV positive serum with a HBV DNA titer of 1.33×108copies/ml was aliquoted into 23 Ep tubes with 1.5 ml, 100 μl in one tube. 15 tubes were incubated at 37 ℃,56 ℃ and 65 ℃ for 0,30,60,120 and 600 min,respectively. The other 8 tubes were incubated at 98 ℃ for 0, 5,10 and 30 min, respectively. Post-treated serum at all time points were selected to infect HepG-2 cell. When 18 hours after infection,these cells were extensively washed with phosphate buffered saline. Cells were harvested after the addition of fresh culture medium to culture cells for 48 hrs. HBV DNA was detected by FQ-PCR. Results HBV DNA was detected in cells that were infected by serum at 37 ℃ and 56 ℃ for 30, 60, 120 and 600 min, respectively. The titers for the cells incubated at 37 ℃ were （4.85±1.71）×105,（3.85±1.76）×105,（1.67±0.67）×105,（7.86±1.03）×104 copies/ml,and those for the cells incubated at 56 ℃ were （4.01±0.16）×105,（9.77±0.97）×104,（6.36±0.65）×104,（5.05±0.24）×103 copies/ml at different incubation time points. For the cells incubated at 65 ℃ for 60 min and 120 min, HBV DNAs were（5.15±7.28）×103 and （7.56±10.6）×102 copies/ml,respectively,which were much lower than those in the controls cells （（6.79±1.48）×105 copies/ml）. The results of HBV DNA were different （Ftreatment×temperature=104.4,P〈0.001） in groups treated with different temperature,and results of HBV DNA were also different （Fprocessing×time=144.0, P〈0.001） in groups processed for different period of time. Temperature and processing time had interaction （Fprocessing time×treatment temperature=23.6, P〈0.001）. After heating at 98 ℃ for 10 min and boiling for 5 min, the HBV DNA copy number（（3.02±4.26）×102,（4.31±6.09）×102 copies/ml） in infected cells decreased by about 10 folds than that in the control group（（6.79±1.48）×105 copies/ml）.HBV DNAs were not detected in cells that were infected by serum which was heated at 98 ℃ for 30 min and boiled for 10 min. Conclusion The infectivity of HBV serum in vitro was relatively stable at low temperture, and it would lose its infectivity in short period of time at high temperature.
出处
《中华预防医学杂志》
CAS
CSCD
北大核心
2011年第8期723-726,共4页
Chinese Journal of Preventive Medicine
基金
中国博士后基金（20090450636）
关键词
肝炎病毒
乙型
热力学
感染控制
评价研究
Hepatitis B virus
Thermodynamics
Infection control
Evaluation studies
