Comparison of virus isolation, immunohistochemistry, and reverse transcriptase-polymerase chain reaction procedures for detection of turkey coronavirus.
A reverse transcriptase-polymerase chain reaction (RT-PCR) procedure and two monoclonal antibody (MAb)-based immunohistochemical procedures were developed for detection of turkey coronavirus (TCV) in tissues and intestinal contents/dropping samples. The RT-PCR, MAb-based fluorescent antibody (FA), and MAb-based immunoperoxidase (IP) procedures were compared with virus isolation (VI) for detection of TCV in experimentally infected turkeys. TCV was detected in experimentally infected turkeys as early as day 1 postexposure (PE) by each of the four detection procedures. TCV was detected as late as day 35 PE by FA or IP and days 42 and 49 PE by VI and RT-PCR, respectively. With VI as a reference, sensitivity and specificity of RT-PCR were 93% and 92%, respectively; specificity of both FA and IP was 96%, and sensitivities were 69% and 61%, respectively. Each of the examined procedures was highly specific, but the RT-PCR procedure was also highly sensitive. These findings demonstrate the utility of both immunohistochemistry and RT-PCR for detection of TCV. In addition, the findings indicate that RT-PCR is a highly sensitive and specific alternative to conventional diagnostic procedures.
Breslin
J J
JJ
Department of Microbiology, Pathology, and Parasitology, North Carolina State University, Raleigh 27606, USA.
Smith
L G
LG
Barnes
H J
HJ
Guy
J S
JS
D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D004752:Coronavirus, Turkey / Q000302:isolation & purification*; D004753:Enteritis, Transmissible, of Turkeys / Q000175:diagnosis*; D005455:Fluorescent Antibody Technique; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry / Q000379:methods; D007413:Intestinal Mucosa / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014422:Turkeys
D000911:Antibodies, Monoclonal