RT-PCR detection of avian coronaviruses of galliform birds (chicken, turkey, pheasant) and in a parrot.
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.
Culver
Francesca Anne
FA
Institute for Animal Health, Compton Laboratory, Newbury, Berkshire, UK.
Britton
Paul
P
Cavanagh
Dave
D
D000818:Animals; D002645:Chickens / Q000821:virology; D017934:Coronavirus / Q000235:genetics* / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D005243:Feces / Q000821:virology; D046094:Galliformes / Q000821:virology*; D010311:Parrots / Q000821:virology*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014422:Turkeys / Q000821:virology
D017931:DNA Primers; D012367:RNA, Viral