TY - JOUR
T1 - A multiplex RT-PCR for the detection of astrovirus, rotavirus, and reovirus in Turkeys
AU - Jindal, Naresh
AU - Chander, Yogesh
AU - Patnayak, Devi P.
AU - Mor, Sunil K.
AU - Ziegler, Andre F.
AU - Goyal, Sagar M.
PY - 2012/9
Y1 - 2012/9
N2 - This study was undertaken to develop and validate a multiplex reverse transcription-polymerase chain reaction (mRT-PCR) for simultaneous detection of avian rotavirus, turkey astrovirus-2 (TAstV-2), and avian reovirus. Primers targeting the conserved regions of NSP4 gene of avian rotavirus, polymerase gene of TAstV-2, and S4 gene of avian reovirus were used. The position of bands at 630, 802, and 1120 base pairs on agarose gel confirmed the presence of rotavirus, TAstV-2, and reovirus, respectively. This mRT-PCR was found to be specific as no amplification was observed with avian influenza virus, Newcastle disease virus, turkey coronavirus, avian metapneumovirus, and intestinal contents of uninfected turkey poults. Intestinal contents of poults from flocks suspected of exhibiting "poult enteritis syndrome" were pooled and tested. Of the 120 pooled samples tested, 70 were positive for TAstV-2, 45 for avian rotavirus, and 18 for avian reovirus. These three viruses were detected alone or in different combinations. Of the samples tested, 20 were negative for these three viruses, 38 were positive for a single virus (TAstV or rotavirus or reovirus), and 42 were positive for two or three viruses. This single-tube mRT-PCR assay has the potential to serve as a rapid diagnostic method for the simultaneous detection of the three enteric viruses in turkeys.
AB - This study was undertaken to develop and validate a multiplex reverse transcription-polymerase chain reaction (mRT-PCR) for simultaneous detection of avian rotavirus, turkey astrovirus-2 (TAstV-2), and avian reovirus. Primers targeting the conserved regions of NSP4 gene of avian rotavirus, polymerase gene of TAstV-2, and S4 gene of avian reovirus were used. The position of bands at 630, 802, and 1120 base pairs on agarose gel confirmed the presence of rotavirus, TAstV-2, and reovirus, respectively. This mRT-PCR was found to be specific as no amplification was observed with avian influenza virus, Newcastle disease virus, turkey coronavirus, avian metapneumovirus, and intestinal contents of uninfected turkey poults. Intestinal contents of poults from flocks suspected of exhibiting "poult enteritis syndrome" were pooled and tested. Of the 120 pooled samples tested, 70 were positive for TAstV-2, 45 for avian rotavirus, and 18 for avian reovirus. These three viruses were detected alone or in different combinations. Of the samples tested, 20 were negative for these three viruses, 38 were positive for a single virus (TAstV or rotavirus or reovirus), and 42 were positive for two or three viruses. This single-tube mRT-PCR assay has the potential to serve as a rapid diagnostic method for the simultaneous detection of the three enteric viruses in turkeys.
KW - avian reovirus
KW - avian rotavirus
KW - multiplex reverse transcription-polymerase chain reaction
KW - poult enteritis syndrome
KW - turkey astrovirus
UR - http://www.scopus.com/inward/record.url?scp=84866726762&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84866726762&partnerID=8YFLogxK
U2 - 10.1637/9958-100911-ResNote.1
DO - 10.1637/9958-100911-ResNote.1
M3 - Article
C2 - 23050480
AN - SCOPUS:84866726762
SN - 0005-2086
VL - 56
SP - 592
EP - 596
JO - Avian diseases
JF - Avian diseases
IS - 3
ER -