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目的比较水体中病毒的三种浓缩方法,评价各方法的浓缩效果,为水体中病毒浓缩方法的选择提供依据。方法将不同浓度的指示病毒(脊髓灰质炎病毒1型PV1)加入经高压灭菌处理后的污水处理厂二级出水中,采用钙离子法、牛肉汤洗膜法和载阳电荷滤料法对水样分别进行浓缩,采用逆转录-PCR技术检测灵敏度,观察指示病毒的某一特征基因片段PCR结果来确定各方法的浓缩效果,并测定回收率。结果钙离子法和载阳电荷滤料法对含最低病毒滴度的水样浓缩后均能检测出特定基因片段,钙离子法的回收率为(91.18±1.77)%,牛肉汤洗膜法的回收率为(85.49±5.59)%,滤料法的回收率为(68.63±2.45)%。结论三种方法均能浓缩水样中较低浓度的肠道病毒,钙离子法回收病毒效果最佳,牛肉汤洗膜法其次,载阳电荷滤料法回收率最低。
Objective To compare the three methods of concentration of virus in water and evaluate the concentration effect of each method and provide the basis for the selection of methods of virus concentration in water. Methods Different concentrations of indicator virus (poliomyelitis virus type 1 PV1) were added to the secondary effluent of the sewage treatment plant after autoclaving. Calcium ion method, beefsteak washing method and cationic charge filter method Water samples were concentrated, the sensitivity of the reverse transcription-PCR assay was used to detect the PCR result of a specific gene fragment indicating the virus to determine the concentration effect of each method, and the recovery rate was determined. Results Calcium ion method and positive charge filter method were able to detect specific gene fragments after concentration of water samples with the lowest virus titer. The recovery rate of calcium ion was (91.18 ± 1.77)%, The recovery rate was (85.49 ± 5.59)%, and the recovery rate of filter method was (68.63 ± 2.45)%. Conclusions All three methods can concentrate the lower concentration of enterovirus in water samples, and the best effect is recovered by calcium ion method. The washing method of beef broth followed by the positive charge filter method has the lowest recovery rate.