Kim Jang Su, Kim Sun Hyung, Bae Sook Young, Lim Chae Seung, Kim Young Kee, Lee Kap No, Lee Chang Kyu
Department of Laboratory Medicine, Korea University College of Medicine, Seoul, Republic of Korea.
J Clin Virol. 2008 Jul;42(3):264-7. doi: 10.1016/j.jcv.2008.03.015. Epub 2008 May 7.
R-Mix, which contains a fresh mixture of two cell lines, Mv1Lu (mink lung cells) and A549 cells, has shown good sensitivity and specificity for respiratory virus culture. However, it has until recently only been available in North America, in part due to the shipping constraints associated with cell aging and the difficulty in providing these cells to hard to reach regions. Recently, cryopreserved R-Mix ReadyCells for longer storage were developed. These cells, which are shipped on dry ice and have a shelf life as long as 6 months from date of manufacture, can be thawed and used as needed with minimal addition of refeeding media.
Assess the potential for cryopreserved R-Mix ReadyCells to replace conventional culture.
Two hundred and twenty-three nasopharyngeal aspirates confirmed as respiratory virus-positive by conventional culture were inoculated into cryopreserved R-Mix ReadyCells and re-inoculated into conventional culture cells simultaneously. After 1 and 3 days of incubation cryopreserved R-Mix ReadyCells and conventional culture cells were screened using a respiratory virus fluorescent antibody pool for the detection of seven major respiratory viruses (influenza A and B viruses, parainfluenza 1, 2 and 3 viruses, respiratory syncytial virus and adenovirus). Positive pool results were further differentiated with specific monoclonal antibodies against the individual viruses.
After 1 day of incubation detection rates for conventional culture were 25%, 39%, 39%, 49%, and 10% for influenza A virus, influenza B virus, parainfluenza viruses, respiratory syncytial virus, and adenovirus, respectively. Corresponding detection rates for cryopreserved R-Mix ReadyCells were 78%, 91%, 72%, 81%, and 65%. Average detection rates of cryopreserved R-Mix ReadyCells for all respiratory viruses were 80% after 1 day incubation and 95% after 3 days incubation, compared to 35% and 70% by conventional culture.
The cryopreserved R-Mix ReadyCells system offers a highly sensitive and rapid method for detection of respiratory viruses that may allow it to replace conventional cell culture systems.
R-Mix包含两种细胞系（Mv1Lu [貂肺细胞]和A549细胞）的新鲜混合物，已显示出对呼吸道病毒培养具有良好的敏感性和特异性。然而，直到最近它仅在北美可用，部分原因是与细胞老化相关的运输限制以及向难以到达的地区提供这些细胞存在困难。最近，开发了用于更长时间储存的冷冻保存的R-Mix ReadyCells。这些细胞通过干冰运输，自制造日期起保质期长达6个月，解冻后可根据需要使用，只需添加极少的再喂养培养基。
评估冷冻保存的R-Mix ReadyCells替代传统培养的潜力。
将223份经传统培养确认为呼吸道病毒阳性的鼻咽抽吸物同时接种到冷冻保存的R-Mix ReadyCells中，并重新接种到传统培养细胞中。在孵育1天和3天后，使用呼吸道病毒荧光抗体池对冷冻保存的R-Mix ReadyCells和传统培养细胞进行筛查，以检测七种主要呼吸道病毒（甲型和乙型流感病毒、副流感1、2和3型病毒、呼吸道合胞病毒和腺病毒）。阳性池结果用针对个别病毒的特异性单克隆抗体进一步区分。
孵育1天后，传统培养对甲型流感病毒、乙型流感病毒、副流感病毒、呼吸道合胞病毒和腺病毒的检出率分别为25%、39%、39%、49%和10%。冷冻保存的R-Mix ReadyCells的相应检出率分别为78%、91%、72%、81%和65%。孵育1天后，冷冻保存的R-Mix ReadyCells对所有呼吸道病毒的平均检出率为80%，孵育3天后为95%，而传统培养分别为35%和70%。
冷冻保存的R-Mix ReadyCells系统提供了一种高度敏感且快速的呼吸道病毒检测方法，这可能使其能够替代传统细胞培养系统。
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