Rapid Fluorescent Detection Assay for Human Parainfluenza Viruses

• Takahashi Tadanobu

  Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka

• Takano Maiko

  Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka

• Kurebayashi Yuuki

  Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka

• Agarikuchi Takashi

  Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka

• Suzuki Chihiro

  Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka

• Fukushima Keijo

  Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka

• Takahashi Shunsaku

  Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka

• Otsubo Tadamune

  Department of Organic Chemistry, School of Pharmaceutical Sciences, Hiroshima International University

• Ikeda Kiyoshi

  Department of Organic Chemistry, School of Pharmaceutical Sciences, Hiroshima International University

• Minami Akira

  Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka

• Suzuki Takashi

  Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka

抄録

Human parainfluenza virus type 1 (hPIV1) does not form clear plaque by the conventional plaque formation assay because of slightly a cytopathic effects in many cell lines infected with hPIV1, thus making in virus titration, isolation and inhibitor evaluation difficult. We have succeeded in fluorescent histochemical visualization of sialidase activities of influenza A and B viruses, Newcastle disease virus and Sendai virus by using a novel fluorescent sialidase substrate, 2-(benzothiazol-2-yl)-4-bromophenyl 5-acetamido-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosidonic acid (BTP3-Neu5Ac). In this study, we applied the BTP3-Neu5Ac assay for rapid detection of hPIV1 and hPIV type 3. The BTP3-Neu5Ac assay could histochemically visualize dot-blotted hPIVs on a membrane and hPIV-infected cells as local fluorescence under UV irradiation. We succeeded in distinct fluorescent visualization of hPIV1-infected cells in only 3 d using the BTP3-Neu5Ac assay. Due to there being no fixation, hPIV1 was isolated directly from fluorescent stained focus cells by the BTP3-Neu5Ac assay. Establishment of a sensitive, easy, and rapid fluorescent focus detection assay for hPIV, hPIV1 in particular will contribute greatly to progress in hPIV studies.

収録刊行物

• Ｂｉｏｌｏｇｉｃａｌ　＆　Ｐｈａｒｍａｃｅｕｔｉｃａｌ　Ｂｕｌｌｅｔｉｎ

  Ｂｉｏｌｏｇｉｃａｌ　＆　Ｐｈａｒｍａｃｅｕｔｉｃａｌ　Ｂｕｌｌｅｔｉｎ 38 (8), 1214-1219, 2015

  公益社団法人 日本薬学会

キーワード

• human parainfluenza virus (hPIV)
• sialidase activity
• fluorescent visualization
• histochemical staining
• 2-(benzothiazol-2-yl)-4-bromophenyl 5-acetamido-3,5-dideoxy-α-D-<i>glycero</i>-D-<i>galacto</i>-2-nonulopyranosidonic acid (BTP3-Neu5A)

詳細情報

• CRID: 1390001204632115200
• NII論文ID: 130005090643
• NII書誌ID: AA10885497
• DOI: 10.1248/bpb.b15-00298
• ISSN: 13475215
• ISSN: 09186158
• NDL書誌ID: 026617333
• PubMed: 26235585
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I026617333
• Web Site: https://www.jstage.jst.go.jp/article/bpb/38/8/38_b15-00298/_pdf
• Web Site: https://search.jamas.or.jp/link/ui/2016128321
• 本文言語コード: en
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