Frank A L, Couch R B, Griffis C A, Baxter B D
J Clin Microbiol. 1979 Jul;10(1):32-6. doi: 10.1128/jcm.10.1.32-36.1979.
Rhesus and cynomolgus monkey kidney tissue cultures and two continuous lines, Madin-Darby canine kidney (MDCK) and LLC-MK2, were compared in titrations and isolations of influenza and parainfluenza viruses. Tube cultures were inoculated with laboratory virus strains or stored patient specimens and observed for hemadsorption. Trypsin was added to the medium of the continuous lines to increase sensitivity. All four tissue cultures gave similar titers of influenza A/USSR (H1N1), A/Texas (H3N2), and B/HK, but lower titers of parainfluenza 1, 2, and 3 were observed with MDCK. Cynomolgus kidney was the best single tissue culture for reisolation of the six viruses, but foamy-virus contamination of many lots was a serious problem. Reisolation of influenza viruses was as successful with MDCK as with primary monkey kidney. LLC-MK2 was similar to rhesus kidney but less successful than cynomolgus kidney. For reisolation of parainfluenza viruses, LLC-MK2 was superior to rhesus monkey kidney and similar to cynomolgus kidney. MDCK was less useful for parainfluenza viruses. Thus, LLC-MK2 would be an acceptable single tissue alternative to primary monkey kidney. The combination of MDCK and LLC-MK2 would provide optimal sensitivity for isolation of all six viruses.
在流感病毒和副流感病毒的滴定和分离实验中,对恒河猴和食蟹猴的肾组织培养物以及两种连续传代细胞系——麦迪逊-达比犬肾细胞(MDCK)和LLC-MK2细胞系进行了比较。将实验室病毒株或保存的患者标本接种到试管培养物中,并观察血细胞吸附情况。向连续传代细胞系的培养基中添加胰蛋白酶以提高敏感性。所有四种组织培养物对甲型流感病毒/苏联(H1N1)、甲型流感病毒/德克萨斯(H3N2)和乙型流感病毒/香港的滴度相似,但MDCK细胞系对副流感病毒1、2和3的滴度较低。食蟹猴肾是重新分离这六种病毒的最佳单一组织培养物,但许多批次存在泡沫病毒污染是一个严重问题。用MDCK细胞系重新分离流感病毒的成功率与原代猴肾细胞相当。LLC-MK2细胞系与恒河猴肾细胞相似,但不如食蟹猴肾细胞成功。对于副流感病毒的重新分离,LLC-MK2细胞系优于恒河猴肾细胞,与食蟹猴肾细胞相似。MDCK细胞系对副流感病毒的作用较小。因此,LLC-MK2细胞系可以作为原代猴肾细胞的一种可接受的单一组织替代物。MDCK细胞系和LLC-MK2细胞系联合使用将为分离所有六种病毒提供最佳敏感性。