模拟水样中甲肝病毒超滤浓缩和PEG浓缩方法的比较  

Comparison of hepatitis A virus concentration methods from simulated water samples using Ultrafiltration and PEG precipitation

作　　者：赵霖[1] 周文亭[1] 曹经瑗 毕胜利[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京102206 [2]通信作者:曹经瑗.Email:[e-mail]

出　　处：《中华实验和临床病毒学杂志》2016年第1期71-75,共5页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的比较模拟水样中甲肝病毒（HAV）超滤浓缩和PEG浓缩方法，为水中HAV检测提供参考。方法向模拟水样中接种已知量的HAV，分别选用Millipore和Sartorius公司的100KD和50KD孔径的微孔超滤管及终浓度为10％聚乙二醇（PEG）-1M氯化钠（NaCl）沉淀的方法对模拟水样进行浓缩，采用荧光定量lit-PCR方法检测浓缩后的病毒量，用SPSS21．0对数据进行统计学分析。结果采用同一孔径不同厂家的超滤管浓缩病毒无统计学差异（P=0．25，P=0．532），对浓缩后的CT值及回收量进行分组比较，结果均表明不同处理方法有统计学差异（P：0．000〈0．05，P=0．000〈0．05），其中50KD组浓缩病毒回收率为（94．57±16．26％），100KD组浓缩病毒回收率为（52．37±14．75％），10％PEG-1．0MNaCl沉淀病毒回收效率为（63．5±8．45％）。结论超滤和PEG沉淀两种方法均可用于水样中HAV浓缩，超滤管浓缩病毒操作简便，耗时更短，选择合适孔径可以提高HAV病毒回收率，本实验中50KD超滤管HAV回收率高于100KD；PEG更适用处理大体积样本。Objective To compare concentration methods of ultrafihration and PEG precipitation for hepatitis A virus（HAV） in simulated water samples, and provide a reference for detection of HAV in water. Methods Quantitative HAV was inoculated in the simulated water samples. Millipore, Sartorius, 100KD and 50KD aperture microporous uhrafihration centrifuge tubes and the final concentration of 10% polyethylene glycol （PEG） -1 M sodium chloride （NaC1） precipitation method were selected to concentrate HAV. The concentration of virus was detected by fluorescence quantitative RT-PCR method. The data were statistically analyzed with SPSS21.0. Results The comparison of CT value and recovery rate shows that there were significant differences in different treatment methods （P 〈 0. 0001, P 〈 0.0001 ）. There were not statistically significant between ultrafiltration centrifuge tubes that they produced by different factory but with the same aperture （P = 0. 25, P = 0. 532）, 50KD group had a higher recovery rate（94. 57± 16. 26% ）than 100KD group （52.37± 14.75%） and the 10%PEG-1.0MNaC1 （63.5 ±8.45%）was in middle of them. Conclusions Both methods of ultrafiltration and PEG precipitation can be used for HAV concentration in water samples, the uhrafiltration centrifuge tube has the advantages of simple operation, less time-consuming and so on, select the appropriate aperture can improve the recovery rate of HAV virus. The HAV recovery rate of 50KD ultraliltration centrifuge tube is higher than 100kD; PEG is more economical and practical, also it can process a large volume sample.

关 键 词：甲肝病毒 病毒浓缩 PEG浓缩 超滤 荧光定量RT-PCR 

分 类 号：R373.21[医药卫生—病原生物学]