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RT-PCR detection of avian coronaviruses of galliform birds (chicken, turkey, pheasant) and in a parrot.

ABSTRACT: 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-checked 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.

SUBMITTER: Culver FA

PROVIDER: S-EPMC7120991 | biostudies-literature | 2008

REPOSITORIES: biostudies-literature

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RT-PCR detection of avian coronaviruses of galliform birds (chicken, turkey, pheasant) and in a parrot.

Culver Francesca Anne FA Britton Paul P Cavanagh Dave D

Methods in molecular biology (Clifton, N.J.) 20080101

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-/UTR4 ...[more]

PMID: 19057865

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