Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix
1 Coronaviruses: An Overview of Their Replication and Pathogenesis . . . . . . . . 1
Anthony R. Fehrand Stanley Perlman
PART I DETECTION, DIAGNOSIS AND EVOLUTION OF CORONAVIRUSES
2 Identification of a Novel Coronavirus from Guinea Fowl
Using Metagenomics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Mariette F. Ducatezand Jean-Luc Guérin
3 Serological Diagnosis of Feline Coronavirus Infection
by Immunochromatographic Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
Tomomi Takanoand Tsutomu Hohdatsu
4 Estimation of Evolutionary Dynamics and Selection Pressure
in Coronaviruses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
Muhammad Munirand Martí Cortey
PART II PROPOGATION, TITRATION, AND PURIFICATION
OF CORONAVIRUSES
5 The Preparation of Chicken Tracheal Organ Cultures
for Virus Isolation, Propagation, and Titration . . . . . . . . . . . . . . . . . . . . . . . . 51
Ruth M. Hennion
6 The Preparation of Chicken Kidney Cell Cultures for Virus Propagation . . . . . 57
Ruth M. Hennionand Gillian Hill
7 Isolation and Propagation of Coronaviruses in Embryonated Eggs . . . . . . . . . 63
James S. Guy
8 Characterization of Human Coronaviruses on Well- Differentiated
Human Airway Epithelial Cell Cultures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
Hulda R. Jonsdottirand Ronald Dijkman
9 Quantification of Infectious Bronchitis Coronavirus
by Titration In Vitro and In Ovo . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89
Joeri Kint , Helena Jane Maier , and Erik Jagt
10 Purification of Coronavirus Virions for Cryo-EM and Proteomic Analysis. . . . 99
Stuart Dentand Benjamin W. Neuman
11 Partial Purification of IBV and Subsequent Isolation of Viral RNA
for Next-Generation Sequencing. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109
Sarah M. Keep , Erica Bickerton , and Paul Britton
Contents
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