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

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

By: Francesca Anne Culver¹, Paul Britton², Dave Cavanagh¹

Abstract

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Of the many primer combinations that we have investigated for the detection of avian coronaviruses, two have worked better than any of the others: they worked with the largest number of strains/samples of a given coronavirus and the most species of avian coronavirus, and they also produced the most sensitive detection tests. The primer combinations were: oligonucleotide pair 2Bp/4Bm, which is in a region of gene 1 that is moderately conserved among all species of coronavirus ( 1 ); and UTR11-/UTR41+, which are in a highly conserved part of the 3′ untranslated region of avian coronaviruses related to infectious bronchitis virus ( 2 ). The gene 1 primer pair enabled the detection of a new coronavirus in a green-cheeked Amazon parrot (Amazon viridigenalis Cassin). In this chapter we describe the use of these oligonucleotides in a one-step (single-tube) RT-PCR, and describe the procedure that we used to extract RNA from turkey feces.

Affiliation(s): (1) Institute for Animal Health, Compton Laboratory, Newbury, Berkshire, UK
(2) Division of Microbiology, Institute for Animal Health, Compton Laboratory, Newbury, Berkshire, UK

Book Title: SARS- and Other Coronaviruses: Laboratory Protocols

Series: Methods in Molecular Biology | Volume: 454 | Year: 2008 | Page Range: 1-8 | DOI: 10.1007/978-1-59745-181-9_4

Subject: Infectious Diseases

Key Words: pan-coronavirus PCRs - coronavirus detection - coronavirus discovery - avian coronaviruses - galliform birds

References

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