用优化的切向流超滤法提高环境水中对虾白斑综合症病毒的浓度及检测效率  

Use of Optimized Tangential Flow Ultrafiltration for Improved Concentration and Detection of White Spot Syndrome Virus in Environmental Water

作　　者：张娜 刘荭 谢艳辉 黄磊 孙思阳 斯泽恩 李家侨 ZHANG Na;LIU Hong;XIE Yan-hui;HUANG Lei;SUN Si-yang;SI Ze-en;LI Jia-qiao(Zhanjiang Customs Technology Center,Zhanjiang 524000,China;Animal and Plant Inspection and Quarantine Technical Centre,Shenzhen Customs,Shenzhen 518045,China;Nanchang Customs Technology Center,Nanchang 330038,China)

机构地区：[1]湛江海关技术中心,广东湛江524000 [2]深圳海关动植物检验检疫技术中心,广东深圳518045 [3]南昌海关技术中心,江西南昌330038

出　　处：《中国兽医杂志》2021年第10期104-110,共7页Chinese Journal of Veterinary Medicine

基　　金：海关总署科技项目(2019HK038,2021HK007);广东出入境检验检疫局科技项目(2018GDK35)。

摘　　要：水是病毒传播的最重要途径之一,但是从水中检测病毒是一项挑战。由于水中的病毒滴度低,而且水的样品成分复杂,检测前通常需要进行水样品处理,浓缩水中的病毒以达到可检测的数量。本试验分析切向流超滤技术的关键参数(包括切向流的不同孔径、不同面积的超滤柱)以提高从水环境样品中浓缩对虾白斑综合症病毒的效率,并用荧光PCR和数字PCR来定量检测病毒浓缩效果。结果显示,浓缩白斑综合症病毒的过滤最佳过滤柱为0.2μm微过滤膜、UFP-100-C-4MA超滤柱(孔径100 kDa,面积为420 cm^(2))。在进行水样品倍比稀释后所构建的切向流超滤方法对白斑综合症病毒的平均浓缩回收率为73.92%,浓缩效果较好;且病毒回收重复性较高。因此,本试验所优化的切向流超滤方法可以有效地浓缩环境水中的虾病毒,提高水生病毒病原体的检测与流行病学调查的效率。Water is one of the most important routes for virus transmission,but the detection of virus in water samples remains a challenge.Because of low virus titer and complex composition,water samples require multistep treatments to ensure sufficient quantity of viruses for detection.This study examined the key parameters(e.g.,ultrafiltration columns with different pore sizes and areas) for tangential flow ultrafiltration to concentrate shrimp white spot syndrome virus(WSSV) from water samples,followed by detection with fluorescence PCR and digital PCR.The results showed that the best filtration column for concentrating WSSV was 0.2 μm microfiltration membrane and UFP-100-C-4 MA ultrafiltration column(pore size of 100 kDa and area of 420 cm^(2)),and that the average concentration recovery rate of the optimized ultrafiltration method for WSSV was 73.92%,with good concentration effect and high repeatability.Thus,the optimized tangential flow ultrafiltration method allows efficient concentration of shrimp virus in environmental water,and improves detection and epidemiological investigation of virus pathogens in aquaculture.

关 键 词：环境水样品 对虾 白斑综合症病毒 切向流超滤 数字PCR 

分 类 号：S855.3[农业科学—临床兽医学]