病毒学实验室的溢洒现象研究

Study on the phenomenon of splashes and sprays from virology

目的研究病毒实验室的实验操作过程中的溢洒现象和病毒液液体跌落后的溅洒范围。方法利用表达绿色荧光蛋白（GFP）的重组腺病毒rADV为模式病毒，通过两组不同工作经验的实验室人员模拟实验室中的常规吸液、移液等操作，通过观察病毒液跌落至96孔细胞培养板是否出现绿色荧光信号来确定实验操作的溢洒情况；我们也将实验常规使用的病毒液量（1ml、1.5ml、4ml、8m1）从实验室可能的两种跌落高度（75cm、110cm）模拟不慎跌落后可能的溅洒范围。结果（1）两组人员使用机械移液装置均未出现溢洒现象；（2）第1组具有5年以上工作经验的实验室人员使用吸管移液等操作，出现因溢洒而导致7孔阳性信号，而2年以内工作经验的第二组人员，在实验操作过程中出现了81孔阳性信号，远大于第1组人员；（3）在两种滴落高度的情况下，1ml、1.5m1、4ml和8ml液体跌落溅撒范围最大半径分别为0.92、1.57、2.63和2.68m。结论生物安全实验室的移液操作过程中实验室人员工作经验非常重要；病毒液不慎跌落的溅撒范围与液体量有关。

Objective To investigate the phenomenon of accidental splashes and sprays from manipulation of recombinant virus material and to measure the approximate spilled distance when recombinant virus material inadvertently dropped in the biosafety laboratory. Methods first, two groups owning different experience simulated the course of accidental spills and splashes by recombinant adenovirus（rADV） which expressed green fluorescence protein（ GFP）, the GFP signal were observed in 96 well cell plate after spills appeared; Second, the routine two heights（75 cm and 110 cm） and capacity（1 ml, 1.5 ml,4 ml and 8 ml） of virus were chose to simulate the experiment of unexpected dropping. Results First, the positive quantity of the first group owning 5 years＇ experience is much less than the second group owning 2 years＇ work experience, the former was 7 positive wells, the latter was 81 positive when they used the pipette to operation. Second, when the unclosed test tubes（ 1 ml, 1.5 nd,4 ml and 8 ml recombinant virus）inadvertently dropped, the largest spill distance was 0.92 m,1.57 m,2.63 m and 2.68 m respectively. Conclusion The better experience is important to make sure safety when we make infectious material; the contaminated distance increased with the amount of recombinant virus material.