水中诺如病毒富集及定量检测研究  

Concentration and Absolute Quantitative Detection of the Norovirus of the Water Specimens

作　　者：周晓红[1,2] 李晖[2] 杨杏芬[2] 柯昌文[2] 陈传德[3] 郑焕英[2] 周惠琼[2] 陈秋霞[2] 邹丽容[2] 

机构地区：[1]暨南大学医学院流行病学教研室,广州510632 [2]广东省疾病预防控制中心,广州510300 [3]深圳市罗湖区疾病预防控制中心,深圳518020

出　　处：《热带医学杂志》2010年第2期137-140,共4页Journal of Tropical Medicine

基　　金：广东省科技计划(No.2007A0300007-7);深圳市罗湖区科技项目(No.2008068)

摘　　要：目的评估MCE-PEG富集法(混合硝酸纤维素膜第一次富集和PEG沉淀第二次富集)对水中诺如病毒的富集效果及检测灵敏度,为水源性诺如病毒胃肠炎疫情暴发的确定提供有效的实验室检测方法。方法把含有诺如病毒的粪便加入纯净水中,用MCE-PEG方法富集后,荧光PCR绝对定量检测富集前后水的病毒浓度,评估该方法的病毒回收率。同时梯度稀释粪便悬液,分别加入纯净水中,评估该方法的灵敏度。结果经过混合硝酸纤维素膜第一次富集后待滤水被浓缩100倍,病毒平均回收率为56.12%,灵敏度是待滤水中病毒浓度为10拷贝/μl。PEG沉淀第二次富集后,浓缩倍数为1000倍,病毒平均回收率为42.47%,灵敏度是待滤水中病毒浓度为1拷贝/μl。结论MCE-PEG富集法操作简单、材料价格实惠、易于购买、病毒回收率及灵敏度较高,可望进一步优化后广泛应用于不同水中病毒富集浓缩检测,为水源性诺如病毒胃肠炎疫情暴发的确定提供了有效的实验室检测方法。Objective To evaluate the MCE-PEG methody in concentrating the norovirus in water and sensitivity of detection for providing a effective laboratory detection method to due with the gastroenteritis outbreak. Methods The feces which contain the norovirus were added to the pure water.The Mixed Cellulose Esters Membrane was used to concentrate the norovirus,followed by PEG concentration.The norovirus in the water specmiens was absolute quantitatively detected by Reahime PCR to evaluate the recovery yields. Also the feces was diluted and added to the pure water samples to evaluate the sensitivity of this method. Results The water specimens which contain the norovirus were concentrated 100 times by Mixed Cellulose Esters Membrane, and the mean recovery yields was 56.12%. The water samples were concentrated 1000 times by PEG, and the mean recovery yields was 42.47%. The sensitivities of detection using the Mixed Cellulose Esters Membrane and PEG to concentrate the virus were 10 copys/μl and 1 copys/μl, respectively. Conclusion The concentration method by Mixed Cellulose Esters Membrane and PEG was appropriate to detect the virus of the water spicemens.This method was simple,and the membrane was inexpensive and easy to purchase, also the recovery yields and the sensitivity were high.

关 键 词：诺如病毒 病毒富集 回收率 灵敏度 

分 类 号：R446.6[医药卫生—诊断学]