摘要
目的通过更换侵染病毒来改进制作“空斑试验”示教板,以得到更鲜明的“空斑试验”结果,使其更好地服务本科实验教学。方法用不同滴度的寨卡病毒和登革病毒感染培养于12孔细胞培养板的Vero细胞,置37℃吸附2 h。吸附后,用1%甲基纤维素Eagle氏覆盖液进行覆盖步骤,置细胞于37℃培养6~7 d将细胞固定和染色后,用自来水冲洗,待干。结果感染寨卡病毒的未死亡脱落的Vero细胞被结晶紫染色,紫色背景明显。肉眼可看到寨卡病毒感染细胞产生的无色空斑,空斑呈圆形,清晰完整,边缘整齐。空斑数量随着病毒浓度的升高而增加,复孔产生空斑数量趋势一致。感染登革病毒的未脱落Vero细胞被染成紫色,形成较鲜明的紫色背景,登革病毒感染细胞产生了无色空斑,但是空斑的形成数量较少,且不是所有复孔都产生了空斑。随着病毒稀释倍数增加,产生的空斑数量无明显减少趋势。结论用寨卡病毒感染Vero细胞能得到更典型的空斑试验结果,该方法更适用于本科教学制作示教结果用。
Objective This experiment aimed to improve the“plaque test”in teaching by replacing the infecting virus to obtain more distinct“plaque test”results,so that it could better serve undergraduate experimental teaching.Methods Infect Vero cell in 12 well cell culture plate was infected with a serial concentration of Zika virus and dengue virus for 2 h in 37℃,and then 1%MC Eagle medium was used for the conducting cover step for 6-7 days at 37℃.After incubation,the infected cells were fixed and stained;cells were then washed with flowing water before observing the results.Results After staining,round,clear and complete plaques with neat edges could be seen and counted by bare eyes under the very bright violet background.The non-dead and exfoliated Vero cells infected with Zika virus were stained with crystal violet,and the purple background was obvious.Colorless plaques produced by Zika virus-infected cells could be seen with the naked eye.The plaques were round,clear and complete,with neat edges.The number of plaques increased with the increase of virus concentration,and the trend of the number of plaques produced in multiple wells was consistent.Unshedded Vero cells infected with dengue virus were stained purple,forming a brighter purple background.Dengue virus-infected cells produced colorless plaques,but the number of plaques formed was small,and were not consistent in duplicate wells.As the virus dilution factor increased,the number of plaques produced had no obvious decreasing trend.Conclusion Infected with Zika virus,a more representative plaque forming test result could be achieved which was more applicable for teaching purpose.
作者
徐鸣阳
朱勋
谭泳谣
谭姹辉
陈剑煌
李美玉
XU Mingyang;ZHU Xun;TAN Yongyao;TAN Chahui;CHEN Jianhuang;LI Meiyu(Basic and Clinical Medicine Teaching Laboratory,School of Medicine,Sun Yat-sen University,Shenzhen,Guangdong 518107,China;Microbiology Laboratory,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou,Guangdong 510080,China;Experimental Teaching Center,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou,Guangdong 510080,China)
出处
《热带医学杂志》
CAS
2024年第1期142-145,151,共5页
Journal of Tropical Medicine
