title: Virus Isolation and Preparation of Sucrose-Banded Chikungunya Virus Samples for Transmission Electron Microscopy.
abstract: Virus isolation and purification is an invaluable technique in virology to detect and characterize viruses. This chapter describes a large-scale Chikungunya virus (CHIKV) propagation and purification methods by using discontinuous sucrose gradient, and sample preparation for transmission electron microscopy. Sucrose-banding yields large quantities of high-titer (10(10) pfu/ml) CHIKV stocks. Such stocks are stable for years when stored at -70 °C.
authors.affiliation: Laboratory of Neurovirology, Department of Virology 1, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuiku-ku, Tokyo, 162-8640, Japan. ck@nih.go.jp.
authors.forename: Chang-Kweng
authors.initials: CK
authors.lastname: Lim
mesh_terms: D000818:Animals; D002499:Centrifugation, Density Gradient / Q000379:methods*; D065632:Chikungunya Fever / Q000097:blood* / Q000821:virology; D002646:Chikungunya virus / Q000254:growth & development / Q000302:isolation & purification*; D002522:Chlorocebus aethiops; D006801:Humans; D046529:Microscopy, Electron, Transmission; D014709:Vero Cells; D019562:Viral Load
keywords: Immunogold labeling; Negative staining; Sucrose gradient; Virus propagation