Detection and isolation of H5N1 influenza virus from large volumes of natural water
Detection and isolation of H5N1 influenza virus from large volumes of natural water
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DOI：
10.1016/j.jviromet.2008.01.001
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发表时间：
2008-04-01
影响因子：
3.1
通讯作者：
Webster, Robert G.
中科院分区：
文献类型：
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作者：
Khalenkov, Alexey;Laver, W. Graeme;Webster, Robert G.
各种水生或湿地鸟类都可能是所有血凝素（HA）亚型禽流感A病毒的天然宿主。病毒进入水中导致水禽之间的传播，是家禽流行病和人类流行病的主要威胁。天然水库中流感病毒的浓度通常太低，无法用大多数方法检测到。该程序旨在检测和分离大量水中的低浓度流感病毒，而无需昂贵的设备和试剂。将病毒吸附到福尔马林固定的红细胞上，随后在鸡胚中分离。使用反向遗传H5 N1病毒确定该方法的灵敏度。有效检测到浓度低至每毫升初始水体积50%鸡蛋感染剂量（EID 50/ml）的0.03。检测概率接近13%，与通过PCR扩增检测流感病毒M基因的概率相当。该方法可用于需要从天然宿主中分离H5 N1流感病毒的简单方法的野外工作者、生态学家、鸟类学家和研究人员。鸡胚中病毒的检测和分离有助于流行病学、遗传学和疫苗研究。(c)2008 Elsevier B. V.保留所有权利。
Various species of aquatic or wetlands birds can be the natural reservoir of avian influenza A viruses of all hemagglutinin (HA) subtypes. Shedding of the virus into water leads to transmission between waterfowl and is a major threat for epidemics in poultry and pandemics in humans. Concentrations of the influenza virus in natural water reservoirs are often too low to be detected by most methods. The procedure was designed to detect and isolate low concentrations of the influenza virus in large volumes of water without the need for costly installations and reagents. The virus was adsorbed onto formalin-fixed erythrocytes and subsequently isolated in chicken embryos. Sensitivity of the method was determined using a reverse-genetic H5N1 virus. A concentration as low as 0.03 of the 50% egg infection dose per milliliter (EID50/ml) of the initial volume of water was effectively detected. The probability of detection was similar to 13%, which is comparable to that of detecting the influenza virus M-gene by PCR amplification. The method can be used by field workers, ecologists, ornithologists, and researchers who need a simple method to isolate H5N1 influenza virus from natural reservoirs. The detection and isolation of virus in embryonated chicken eggs may help epidemiologic, genetic, and vaccine studies. (c) 2008 Elsevier B.V. All rights reserved.