Krauss Scott, Walker David, Webster Robert G
Division of Virology, Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, TN, USA.
Methods Mol Biol. 2012;865:11-24. doi: 10.1007/978-1-61779-621-0_2.
The isolation of influenza viruses is important for the diagnosis of respiratory diseases in lower animals and humans, for the detection of the infecting agent in surveillance programs, and is an essential element in the development and production of vaccine. Since influenza is caused by a zoonotic virus it is necessary to do surveillance in the reservoir species (aquatic waterfowls), intermediate hosts (quails, pigs), and in affected mammals including humans. Two of the hemagglutinin (HA) subtypes of influenza A viruses (H5 and H7) can evolve into highly pathogenic (HP) strains for gallinaceous poultry; some HP H5 and H7 strains cause lethal infection of humans. In waterfowls, low pathogenic avian influenza (LPAI) isolates are obtained primarily from the cloaca (or feces); in domestic poultry, the virus is more often recovered from the respiratory tract than from cloacal samples; in mammals, the virus is most often isolated from the respiratory tract, and in cases of high pathogenic avian influenza (HPAI) from the blood and internal organs of infected birds. Virus isolation procedures are performed by inoculation of clinical specimens into embryonated eggs (primarily chicken eggs) or onto a variety of primary or continuous tissue culture systems. Successful isolation of influenza virus depends on the quality of the sample and matching the appropriate culture method to the sample type.
流感病毒的分离对于诊断低等动物和人类的呼吸道疾病、在监测项目中检测感染因子以及疫苗的研发和生产至关重要。由于流感是由一种人畜共患病毒引起的,因此有必要在储存宿主物种(水禽)、中间宿主(鹌鹑、猪)以及包括人类在内的受感染哺乳动物中进行监测。甲型流感病毒的两种血凝素(HA)亚型(H5和H7)可演变成对鸡形目家禽具有高致病性(HP)的毒株;一些高致病性H5和H7毒株可导致人类致命感染。在水禽中,低致病性禽流感(LPAI)分离株主要从泄殖腔(或粪便)中获得;在家禽中,病毒更多地从呼吸道而非泄殖腔样本中分离出来;在哺乳动物中,病毒最常从呼吸道分离出来,在高致病性禽流感(HPAI)病例中,则从受感染鸟类的血液和内脏中分离出来。病毒分离程序是通过将临床标本接种到鸡胚(主要是鸡蛋)或各种原代或连续组织培养系统中来进行的。流感病毒的成功分离取决于样本质量以及使适当的培养方法与样本类型相匹配。