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目的比较两相分离法和阴离子滤膜吸附法在浓缩处理外环境污水脊髓灰质炎病毒时的回收效率,选择更高效、便捷的方法以提高外环境污水脊灰病毒监测的敏感性。方法在灭菌的外环境污水中人工添加I型脊灰病毒,使之总病毒量分别达到10~4、10~3、10~2半数组织培养感染剂量(TCID_(50)),再分别用两相分离法和阴离子滤膜吸附法对污水中的脊灰病毒进行浓缩,用实时荧光定量PCR方法和细胞培养方法测定回收病毒的浓度。结果使用实时荧光定量PCR方法测定病毒添加量为10~4TCID_(50)时两相分离法的病毒回收率高于阴离子滤膜吸附法;细胞培养方法测定结果为添加10~4TCID_(50)病毒量时两相分离法和阴离子滤膜吸附法的病毒回收率分别为44.78%和41.06%。结论在对存在脊灰病毒的外环境监测中,两相分离法的浓缩效果整体优于阴离子滤膜吸附法,适合长期应用。
OBJECTIVE To compare the recovery efficiency of two-phase separation and anion-exchange membrane adsorption in the enrichment of poliovirus polluted by external environment pollutions, and select a more efficient and convenient method to improve the sensitivity of external poliovirus monitoring. Methods The poliovirus type I was artificially added to the sterilized external environment water so that the total viral load reached 10 ~ 4, 10 ~ 3 and 10 ~ 2 half of the tissue culture infection dose (TCID_ (50)), respectively Two-phase separation method and anion filter adsorption method for polluting the poliovirus in sewage, using real-time fluorescence quantitative PCR method and cell culture method to determine the concentration of recovered virus. Results The real-time PCR method was used to determine the virus recovery of the two-phase separation method when the virus dosage was 10 ~ 4TCID_ (50) compared with that of the anion-exchange membrane adsorption method. The cell culture method was determined by adding 10 ~ 4TCID_ (50) When the two-phase separation and anion exchange membrane adsorption of the virus recovery rates were 44.78% and 41.06%. Conclusion In the external environment monitoring of poliovirus, the two-phase separation method is better than the anion-exchange membrane adsorption method in overall concentration, which is suitable for long-term application.