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Title: 同時感染したHEK細胞を用いたアデアウイルス5型のプラーク法
Other Titles: Plaque Assay Method for Adenovirus Type 5 with the Culture of HEK Cells Synchronously Infected with the Virus.
Authors: 上田, 芳秋 / 明石, 光伸 / 林, 薫
Authors (alternative): Ueda, Yoshiaki / Akashi, Mitsunobu / Hayashi, Kaoru
Issue Date: 30-Jan-1976
Publisher: 長崎大学熱帯医学研究所 / Institute of Tropical Medicine, Nagasaki University
Citation: 熱帯医学 Tropical medicine 17(3). p151-158, 1976
Abstract: 適宜に階段稀釈した抗血清とX×10^4 PFUのウイルスを混じ室温1時間放置した後,直ちに1:100まで稀釈する.この稀釈液と等量の維持に浮游させた2~3×10^6/mlのHEK細胞とを混和し,37℃30分間軽く振盪する.径5cmのシヤレーに1mlずつ入れ,細胞をガラス面に拡げ,0.5%仔牛血清,0.75%カルボオキシメチールセルローズを含んだ維持液を加え蔽う.5日ないし6日目に細胞をギムザ液で染色し,プラークを算える.以上が抗血清の中和抗体の測定法である.抗血清と抗原(分画その他)を加え充分反応させた後,更らにX×10^4PFUのウイルスを追加し室温60分放置し,以後の手順は上記の方法に従う,以上が抗原による抗血清中の抗体のブロック能を知る方法である.以上の方法を用いて,アデノウイルス5型の抗血清に対するA抗原,C抗原及びP抗原のブロック能を検査した.抗血清が完全にブロックされたのはA抗原によってのみであり, C抗原及びP抗原は抗血清のブロック能を有しないことを知った. / The mixture of antiserum and antigen at each adequate dilution was added the seed virus contained X×10^4 PFU per ml. The mixture was diluted upto the concentration contained 50 to 100 plaques per ml per a dish and 2 ml of the last dilution was made. The last dilution of the mixture was added equal volume of HEK cell suspension contained 2 or 3×10^6 cells per ml and shaked at 37℃ for 20 or 30 minutes. One ml of these mixture of antiserum, antigen and seed virus was plated into a dish, and 4 ml of maintainance medium contained 0.5% calf serum and 0.75% carboxymethylcellulose was added and spread over in a dish. After incubation for 5 or 6 days, the cell sheet was fixed with 10% formaldehyde saline solution and washed and stained with Gimsa solution. The plaques formed in this method were clear and easy to count. The activity of antigen A, C and P of adenovirus type 5 for the blocking antibody against purified adenovirus type 5 was studied with the application of this plaque assay method.
URI: http://hdl.handle.net/10069/4188
ISSN: 03855643
Type: Departmental Bulletin Paper
Appears in Collections:Volume 17, No. 3

Citable URI : http://hdl.handle.net/10069/4188

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