SARS- and Other Coronaviruses

Laboratory Protocols

Editors:

ISBN: 978-1-58829-867-6 (Print) 978-1-59745-181-9 (Online)

Table of contents (23 protocols)

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  1. Front Matter

    Pages i-xv

  2. Detection and Discovery of Coronaviruses

    1. Front Matter

      Pages 1-1

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      Pages 3-12

      A Pancoronavirus RT-PCR Assay for Detection of All Known Coronaviruses

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      Pages 13-26

      Detection of Group 1 Coronaviruses in Bats Using Universal Coronavirus Reverse Transcription Polymerase Chain Reactions

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      Pages 27-34

      Detection and Sequence Characterization of the 3′-End of Coronavirus Genomes Harboring the Highly Conserved RNA Motif s2m

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      Pages 35-42

      RT-PCR Detection of Avian Coronaviruses of Galliform Birds (Chicken, Turkey, Pheasant) and in a Parrot

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      Pages 43-59

      Detection of Group 2a Coronaviruses with Emphasis on Bovine and Wild Ruminant Strains

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      Pages 61-72

      Detection of SARS Coronavirus in Humans and Animals by Conventional and Quantitative (Real Time) Reverse Transcription Polymerase Chain Reactions

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      Pages 73-89

      Detection of New Viruses by VIDISCA

  3. Isolation, Growth, Titration, and Purification of Coronaviruses

    1. Front Matter

      Pages 91-91

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      Pages 93-102

      Titration of Human Coronaviruses, HCoV-229E and HCoV-OC43, by an Indirect Immunoperoxidase Assay

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      Pages 103-107

      The Preparation of Chicken Tracheal Organ Cultures for Virus Isolation, Propagation, and Titration

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      Pages 109-117

      Isolation and Propagation of Coronaviruses inEmbryonated Eggs

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      Pages 119-126

      Large-Scale Preparation of UV-Inactivated SARS Coronavirus Virions for Vaccine Antigen

  4. Structure of Covs Analyzed by Electron Microscopy

    1. Front Matter

      Pages 127-127

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      Pages 129-136

      Purification and Electron Cryomicroscopy of Coronavirus Particles

  5. Expression of Coronavirus Proteins and Crystallization

    1. Front Matter

      Pages 137-137

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      Pages 139-159

      Production of Coronavirus Nonstructural Proteins in Soluble Form for Crystallization

  6. Raising Antibodies to Coronavirus Proteins

    1. Front Matter

      Pages 161-161

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      Pages 163-189

      Generating Antibodies to the Gene 3 Proteins of Infectious Bronchitis Virus

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      Pages 191-203

      Establishment and Characterization of Monoclonal Antibodies Against SARS Coronavirus

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      Pages 205-226

      Production of Monospecific Rabbit Antisera Recognizing Nidovirus Proteins

  7. Manipulating the Genomes of Coronaviruses

    1. Front Matter

      Pages 227-227

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      Pages 229-236

      Manipulation of the Coronavirus Genome Using Targeted RNA Recombination with Interspecies Chimeric Coronaviruses

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      Pages 237-254

      Generation of Recombinant Coronaviruses Using Vaccinia Virus as the Cloning Vector and Stable Cell Lines Containing Coronaviral Replicon RNAs

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      Pages 255-273

      Transient Dominant Selection for the Modification and Generation of Recombinant Infectious Bronchitis Coronaviruses

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      Pages 275-291

      Engineering Infectious cDNAs of Coronavirus as Bacterial Artificial Chromosomes

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