By Yoshihiro Kawaoka, Gabriele Neumann
Reports of influenza-like health problems date again to the center a long time, and outbreaks of influenza most likely troubled people lengthy earlier than that. over the past part century, influenza virus study has ended in the advance of 2 periods of antivirals – ion channel and neuraminidase inhibitors. lately, a mode of the bogus iteration of an influenza virus was once verified. the program has been instrumental within the improvement of novel influenza vaccines and within the figuring out of viral pathogenicity and the features of viral proteins. Influenza Virus: tools and Protocols summarizes the present suggestions that experience made this development attainable, starting from protocols for virus isolation, development, and subtyping to approaches for the effective iteration of any influenza virus. Written within the winning Methods in Molecular Biology™ sequence layout, chapters contain introductions to their respective issues, lists of the mandatory fabrics and reagents, step by step, quite simply reproducible protocols, and notes on troubleshooting and averting recognized pitfalls.
Authoritative and simply obtainable, Influenza Virus: equipment and Protocols seeks to serve either execs and newcomers with the suggestions utilized in a variety of laboratories all over the world which are, hence, the construction blocks that underpin just about all influenza virus examine.
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Extra resources for Influenza Virus: Methods and Protocols
J Virol. 77, 8418–25. 21. C. (1998) New helpercells and matched early region 1-deleted adenovirus vectors prevent generation of replication -competent adenoviruses. Hum Gene Ther. 9, 1909–17. 22. , Blödorn, J. (1975) Activation of influenza A viruses by trypsin treatment. Virology. 68, 426–39. 23. L. G. (2010) Perspectives on influenza evolution and the role of research. Anim Health Res Rev. 11, 3–18. Chapter 3 Influenza Virus Titration, Antigenic Characterization, and Serological Methods for Antibody Detection Alexander Klimov, Amanda Balish, Vic Veguilla, Hong Sun, Jarad Schiffer, Xiuhua Lu, Jacqueline M.
3. Stack plates and cover with an empty plate. Place in 37°C, 5% CO2 incubator for 1 h to mimic the virus + serum incubation step of the microneutralization assay. 4. Prepare MDCK cells for use in assay as described. Add 100 ml of diluted cells to each well of the microtiter plate. 5 × 104 cells. Incubate at 37°C in 5% CO2 for 18–20 h. 5. The procedures for day 2 of the TCID determination, plate fixation, 1° antibody, 2° antibody, and substrate, are the same as these procedures for the microneutralization assay.
5004). 3. MDCK cells (ATCC# CCL-34) confluent in a 6-well tissue culture plates (Nunc, Thermo-Fisher Scientific, Cat. # 150239). 4. Plaque assay wash medium: supplement 490 ml of DMEM with 5 ml of penicillin–streptomycin (100 U/ml penicillin G and 100 mg/ml streptomycin). 2-mm membrane filter. Can be stored up to 2 months at 4–8°C. 5. L-1-Tosylamide-2-phenylethyl chloromethyl ketone (TPCK)– trypsin (2 mg/ml) working solution: supplement 10 ml of *Identification of particular products is provided as a guide to aid in the selection of equivalent, suitable products 30 A.