WHO Collaborating Centre for Reference and Research on Influenza, Victorian Infectious Diseases Reference Laboratory, at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, 3000, Australia.
WHO Collaborating Centre for Reference and Research on Influenza, Victorian Infectious Diseases Reference Laboratory, at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, 3000, Australia.
J Clin Virol. 2017 Dec;97:54-58. doi: 10.1016/j.jcv.2017.10.018. Epub 2017 Nov 2.
The isolation and propagation of influenza viruses from clinical specimens are essential tools for comprehensive virologic surveillance. Influenza viruses must be amplified in cell culture for detailed antigenic analysis and for phenotypic assays assessing susceptibility to antiviral drugs or for other assays.
To conduct an external quality assessment (EQA) of proficiency for isolation and identification of influenza viruses using cell culture techniques among National Influenza Centres (NICs) in the World Health Organisation (WHO) South East Asia and Western Pacific Regions.
Twenty-one NICs performed routine influenza virus isolation and identification techniques on a proficiency testing panel comprising 16 samples, containing influenza A or B viruses and negative control samples. One sample was used exclusively to determine their capacity to measure hemagglutination titer and the other 15 samples were used for virus isolation and identification.
All NICs performed influenza virus isolation using Madin Darby canine kidney (MDCK) or MDCK-SIAT-1 cells. If virus growth was detected, the type, subtype and/or lineage of virus present in isolates was determined using immunofluorescence, RT-PCR and/or hemagglutination inhibition (HI) assays. Most participating laboratories could detect influenza virus growth and could identify virus amplified from EQA samples. However, some laboratories failed to isolate and identify viruses from EQA samples that contained lower titres of virus, highlighting issues regarding the sensitivity of influenza virus isolation methods between laboratories.
This first round of EQA was successfully conducted by NICs in the Asia Pacific Region, revealing good proficiency in influenza virus isolation and identification.
从临床标本中分离和培养流感病毒是全面进行病毒学监测的重要工具。为了进行详细的抗原分析以及评估对抗病毒药物的敏感性或其他检测，流感病毒必须在细胞培养中扩增。
通过细胞培养技术，在世界卫生组织（WHO）东南亚和西太平洋地区的国家流感中心（NIC）中进行流感病毒分离和鉴定能力的外部质量评估（EQA）。
21 个 NIC 对一个包含 16 个样本的能力测试面板进行了常规的流感病毒分离和鉴定技术，这些样本包含了甲型或乙型流感病毒以及阴性对照样本。一个样本专门用于确定其测定血凝滴度的能力，而另外 15 个样本用于病毒分离和鉴定。
所有 NIC 都使用 Madin Darby 犬肾（MDCK）或 MDCK-SIAT-1 细胞进行流感病毒分离。如果检测到病毒生长，则使用免疫荧光、RT-PCR 和/或血凝抑制（HI）检测来确定分离物中存在的病毒的类型、亚型和/或谱系。大多数参与实验室能够检测到流感病毒的生长，并能够鉴定从 EQA 样本中扩增的病毒。然而，一些实验室未能从 EQA 样本中分离和鉴定出病毒，这些样本中病毒滴度较低，这突出了实验室之间流感病毒分离方法的敏感性问题。
亚太地区的 NIC 成功地进行了第一轮 EQA，显示出在流感病毒分离和鉴定方面的良好能力。
