不同培养基分离流感病毒效果及最佳培养条件

Effect of different culture medium in isolating influenza virus and exploration of the most ideal culture conditions

目的比较有血清培养基与无血清培养基分离流感病毒的效果,探索无血清培养基分离流感病毒的最佳条件。方法将原始标本接种于2种培养基培养的MDCK细胞,比较流感病毒分离效果。将原始标本接种于无血清培养基培养的MDCK细胞,比较MDCK细胞、样品液、接种方法、胞龄、培养温度、无血清病毒分离液含TPCK-胰酶浓度对流感病毒的影响,确定最优培养条件,红细胞凝集试验测血凝滴度。结果 2种培养基分离流感病毒的阳性率差异与血凝滴度差异有统计学意义(P<0.05);无血清培养基的细胞不洗涤、24 h与48 h胞龄、直接接种、2μg/ml^3μg/ml TPCK-胰酶、34.5℃培养条件下病毒血凝滴度较高。结论无血清培养基分离流感病毒优于有血清培养基且操作简便。

Objective The effects of serum culture medium and serum free media were compared for influenza virus, and the optimal conditions for the influenza virus isolation in serum free medium were explored. Methods The original samples were respectively inoculated in MDCK cells with two kinds of culture medium, and the effect of influenza virus isolation was compared. The original samples were respectively inoculated in MDCK cells of serum free medium, and by comparing cells of MDCK, sample solution（ discard, leave behind）, inoculation methods, ceUular age, culture temperature, the effect of TPCK pancreatic enzyme dosage by serum free medium that were influenced on influenza virus, the best culture conditions was determined. Hemagglutinatio method was used to detect the virus titers. Results Through the comparison of temagglutinatio method with culture medium to isolate, the differences of positive rate and hemagglutination titer of influenza virus isolations between the two medium were statistically significant （ P 〈 0.05 ）. The virus hemagglutination titer of serum free medium was higher in the culture conditions of no cells washing, cell ages at 24 h and 48 h, direct inoculation, 2μg/ml 3 μg/ml TPCK pancreatin, at 34.5℃. Conclusion Serum -free medium is easier in operation than serum medium in isolating influenza virus.