CAJ | 학술논문

目的建立饮用水中病毒浓缩与检测方法并对实际水样进行人腺病毒污染状况监测.方法以f2噬菌体为水中肠道病毒代表,投加于受试水样中,评价NanoCeram滤芯对原水和饮用水初次浓缩及不同浓度PEG 8000对初次浓缩液进行二次浓缩的效果.采用T-A克隆制备人腺病毒荧光定量PCR标准品,对所得质粒测序,应用Blast序列类似性检索工具将其与目的基因片段进行一致性检测,建立荧光定量PCR检测人腺病毒的方法.应用上述方法,于2011年对武汉两家自来水厂原水和饮用水采用NanoCeram滤芯进行现场初次浓缩,聚乙二醇(PEG)/NaCl法进行二次浓缩,提取浓缩液中病毒DNA后进行荧光定量PCR检测,监测原水和饮水中人腺病毒污染状况.结果所建立的NanoCeram滤芯初次浓缩法对原水中的f2噬菌体回收率为(51.63±26.60)％,对饮用水中的f2噬菌体回收率为(50.27±14.35)％.当PEG 8000浓度达到0.13 kg/L时,二次浓缩回收率可达(90.09±10.50)％.所构建的人腺病毒荧光定量PCR标准品与目的基因片段序列一致度为99％,可用于人腺病毒的绝对定量.2011年,武汉市两家自来水厂原水中的人腺病毒浓度范围为(4.13×103～2.20×106)拷贝/L,出厂水中的人腺病毒浓度范围为(5.57×102 ～7.52×1O5)拷贝/L,饮用水处理过程对人腺病毒的清除率为(75.49±11.71)％.结论 NanoCeram滤芯联合PEG/NaCl法可对水环境中病毒进行有效浓缩,应用所建立的荧光定量PCR法检测发现自来水厂原水中存在人腺病毒,目前的水处理措施不能将其完全去除.
목적 건립음용수중병독농축여검측방법병대실제수양진행인선병독오염상황감측.방법 이f2서균체위수중장도병독대표,투가우수시수양중,평개NanoCeram려심대원수화음용수초차농축급불동농도PEG 8000대초차농축액진행이차농축적효과.채용T-A극륭제비인선병독형광정량PCR표준품,대소득질립측서,응용Blast서렬유사성검색공구장기여목적기인편단진행일치성검측,건립형광정량PCR검측인선병독적방법.응용상술방법,우2011년대무한량가자래수엄원수화음용수채용NanoCeram려심진행현장초차농축,취을이순(PEG)/NaCl법진행이차농축,제취농축액중병독DNA후진행형광정량PCR검측,감측원수화음수중인선병독오염상황.결과 소건립적NanoCeram려심초차농축법대원수중적f2서균체회수솔위(51.63±26.60)％,대음용수중적f2서균체회수솔위(50.27±14.35)％.당PEG 8000농도체도0.13 kg/L시,이차농축회수솔가체(90.09±10.50)％.소구건적인선병독형광정량PCR표준품여목적기인편단서렬일치도위99％,가용우인선병독적절대정량.2011년,무한시량가자래수엄원수중적인선병독농도범위위(4.13×103～2.20×106)고패/L,출엄수중적인선병독농도범위위(5.57×102 ～7.52×1O5)고패/L,음용수처리과정대인선병독적청제솔위(75.49±11.71)％.결론 NanoCeram려심연합PEG/NaCl법가대수배경중병독진행유효농축,응용소건립적형광정량PCR법검측발현자래수엄원수중존재인선병독,목전적수처리조시불능장기완전거제.
Objective This study aimed to construct an effective method to concentrate and detect virus in drinking water,and human adenovirus pollution status in actual water samples was monitored by constructed method.Methods The concentration efficient of NanoCeram filter for the first concentration with source water and drinking water and the concentration efficient of the different concentrations of PEG 8000 for the second concentration were assessed by spiking f2 bacteriophage into water samples.The standard of human adenovirus for real-time PCR was constructed by T-A clone.The plasmid obtained was identified through sequence analyzing and consistency check comparing to target gene fragment was conducted by using blast algorithm.Then,real-time PCR was constructed to quantify the concentration of human adenovirus using the plasmid as standard.Water samples were concentrated by using NanoCeram filter on the spot and then concentrated for the second time by PEG/NaCl in 2011.The DNA of concentrated samples were extracted for the quantification of human adenovirus in real-time PCR subsequently to monitor the pollution of human adenovirus in water.Results For the first concentration by NanoCeram filter,the recovery rates were (51.63 ±26.60) ％ in source water and (50.27 ± 14.35) ％ in treated water,respectively.For the second concentration,the highest recovery rate was reached to (90.09 ± 10.50 ) ％ at the concentration of 0.13 kg/L of PEG 8000.The sequence identity score of standard of adenovirus for real time PCR and adenovirus gene was 99％,implying that it can be successfully used to quantification with human adenovirus.The levels of human adenovirus in the water samples sampled in 2011 ranged from 4.13 × 103 to 2.20 × 106 copies/L in source water,while range from 5.57 × 102 to 7.52 × 105 copies/L in treated water and the removal efficiency range was (75.49 ± 11.71 )％.Conclusion NanoCeram filers combined with PEG/NaCl was an effective method to concentrate virus in aquatic environment.There was a large number of human adenovirus in source water,and it is not sufficient to remove them thoroughly through conventional water treatment processes.