@article{guy_smith_breslin_pakpinyo_2002, title={Development of a competitive enzyme-linked immunosorbent assay for detection of turkey coronavirus antibodies}, volume={46}, ISSN={["0005-2086"]}, DOI={10.1637/0005-2086(2002)046[0334:DOACEL]2.0.CO;2}, abstractNote={SUMMARY. A competitive enzyme-linked immunosorbent assay (cELISA) was developed for detection of turkey coronavirus (TCV) antibodies. The cELISA utilized a recombinant baculovirus (Autographa californica nuclear polyhedrosis virus)-expressed TCV nucleocapsid (N) protein and biotin-labeled TCV N protein-specific monoclonal antibody. Sensitivity and specificity of the cELISA for detection of TCV antibodies were determined by comparison with the indirect fluorescent antibody test (IFAT) with 1269 reference, experimentally derived, and field-origin sera. Sera with discordant cELISA and IFAT results were further evaluated by western immunoblot analyses. The cELISA detected antibodies specific for TCV and infectious bronchitis virus, a closely related coronavirus, but did not detect antibodies specific for other avian viruses. A high degree of concordance was observed between the cELISA and IFAT; sensitivity and specificity of the cELISA relative to IFAT were 92.9% and 96.2%, respectively. Western immunoblot analyses provided additional evidence of cELISA specificity. The findings indicate that the cELISA is a rapid, sensitive, and specific serologic test for detection of TCV antibodies in turkeys.}, number={2}, journal={AVIAN DISEASES}, author={Guy, JS and Smith, LG and Breslin, JJ and Pakpinyo, S}, year={2002}, pages={334–341} }
 @article{breslin_smith_guy_2001, title={Baculovirus expression of turkey coronavirus nucleocapsid protein}, volume={45}, ISSN={["0005-2086"]}, DOI={10.2307/1593020}, abstractNote={The nucleocapsid (N) gene of turkey coronavirus (TCV) was amplified by reverse transcriptase-polymerase chain reaction, cloned, and expressed in the baculovirus expression system. A recombinant baculovirus containing the TCV N gene (rBTCV/N) was identified by polymerase chain reaction and expression of TCV N protein as determined by western immunoblot analysis. Two TCV-specific proteins, 52 and 43 kDa, were expressed by rBTCV/N; one of these proteins, p52, was comparable in size to native TCV N protein. Baculovirus-expressed N proteins were used as antigen in an indirect enzyme-linked immunosorbent assay (ELISA) for detection of TCV-specific antibodies. The ELISA detected antibodies specific for TCV and infectious bronchitis virus, a closely related avian coronavirus, but did not detect antibodies specific for other avian viruses (avian influenza, avian reovirus, avian paramyxovirus 3, avian adenovirus 1, or Newcastle disease virus). These findings indicate that baculovirus-expressed TCV N protein is a suitable source of antigen for ELISA-based detection of TCV-specific antibodies in turkeys.}, number={1}, journal={AVIAN DISEASES}, author={Breslin, JJ and Smith, LG and Guy, JS}, year={2001}, pages={136–143} }
 @article{guy_breslin_breuhaus_vivrette_smith_2000, title={Characterization of a coronavirus isolated from a diarrheic foal}, volume={38}, number={12}, journal={Journal of Clinical Microbiology}, author={Guy, J. S. and Breslin, J. J. and Breuhaus, B. and Vivrette, S. and Smith, L. G.}, year={2000}, pages={4523–4526} }
 @article{breslin_smith_barnes_guy_2000, title={Comparison of virus isolation, immunohistochemistry, and reverse transcriptase-polymerase chain reaction procedures for detection of turkey coronavirus}, volume={44}, ISSN={["1938-4351"]}, DOI={10.2307/1593102}, abstractNote={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.}, number={3}, journal={AVIAN DISEASES}, author={Breslin, JJ and Smith, LG and Barnes, HJ and Guy, JS}, year={2000}, pages={624–631} }
 @article{guy_smith_breslin_vaillancourt_barnes_2000, title={High mortality and growth depression experimentally produced in young turkeys by dual infection with enteropathogenic Escherichia coli and turkey coronavirus}, volume={44}, ISSN={["0005-2086"]}, DOI={10.2307/1592513}, abstractNote={Six-day-old turkeys were inoculated with turkey coronavirus (TCV) and an enteropathogenic Escherichia coli (EPEC) (isolate R98/5) that were isolated from poult enteritis and mortality syndrome (PEMS)-affected turkeys. Turkeys inoculated with only R98/5 did not develop clinically apparent disease, and only mild disease and moderate growth depression were observed in turkeys inoculated with only TCV. Turkeys dually inoculated with TCV and R98/5 developed severe enteritis with high mortality (38/48, 79%) and marked growth depression. R98/5 infection resulted in attaching/effacing (AE) intestinal lesions characteristic of EPEC: adherence of bacterial microcolonies to intestinal epithelium with degeneration and necrosis of epithelium at sites of bacterial attachment. AE lesions were more extensive and were detected for a prolonged duration in dually inoculated turkeys compared with turkeys inoculated with only R98/5. An apparent synergistic effect in dually inoculated turkeys was indicated by increased mortality, enhanced growth depression, and enhanced AE lesion development. The results suggest that TCV promoted intestinal colonization by R98/5; however, R98/5 did not appear to alter TCV infection. The present study provides a possible etiologic explanation for PEMS.}, number={1}, journal={AVIAN DISEASES}, author={Guy, JS and Smith, LG and Breslin, JJ and Vaillancourt, JP and Barnes, HJ}, year={2000}, pages={105–113} }
 @article{breslin_smith_fuller_guy_1999, title={Sequence analysis of the matrix nucleocapsid gene region of turkey coronavirus}, volume={42}, ISSN={["0300-5526"]}, DOI={10.1159/000024956}, abstractNote={A reverse transcriptase, polymerase chain reaction (RT-PCR) procedure was used to amplify a segment of the genome of turkey coronavirus (TCV) spanning portions of the matrix and nucleocapsid (MN) protein genes (approximately 1.1 kb). The MN gene region of three epidemiologically distinct TCV strains (Minnesota, NC95, Indiana) was amplified, cloned into pUC19, and sequenced. TCV MN gene sequences were compared with published sequences of other avian and mammalian coronaviruses. A high degree of similarity (>90%) was observed between the nucleotide, matrix protein, and nucleocapsid protein sequences of TCV strains and published sequences of infectious bronchitis virus (IBV). The matrix and nucleocapsid protein sequences of TCV had limited homology (<30%) with MN sequences of mammalian coronaviruses. These results demonstrate a close genetic relationship between the avian coronaviruses, IBV and TCV.}, number={1}, journal={INTERVIROLOGY}, author={Breslin, JJ and Smith, LG and Fuller, FJ and Guy, JS}, year={1999}, pages={22–29} }
 @article{breslin_smith_fuller_guy_1999, title={Sequence analysis of the turkey coronavirus nucleocapsid protein gene and 3 ' untranslated region identifies the virus as a close relative of infectious bronchitis virus}, volume={65}, ISSN={["0168-1702"]}, DOI={10.1016/S0168-1702(99)00117-3}, abstractNote={The 3' end of the turkey coronavirus (TCV) genome (1740 bases) including the nucleocapsid (N) gene and 3' untranslated region (UTR) were sequenced and compared with published sequences of other avian and mammalian coronaviruses. The deduced sequence of the TCV N protein was determined to be 409 amino acids with a molecular mass of approximately 45 kDa. The TCV N protein was identical in size and had greater than 90% amino acid identity with published N protein sequences of infectious bronchitis virus (IBV); less than 21% identity was observed with N proteins of bovine coronavirus and transmissible gastroenteritis virus. The 3' UTR showed some variation among the three TCV strains examined, with two TCV strains, Minnesota and Indiana, containing 153 base segments which are not present in the NC95 strain. Nucleotide sequence identity between the 3' UTRs of TCV and IBV was greater than 78%. Similarities in both size and sequence of TCV and IBV N proteins and 3' UTRs provide additional evidence that these avian coronaviruses are closely related.}, number={2}, journal={VIRUS RESEARCH}, author={Breslin, JJ and Smith, LG and Fuller, FJ and Guy, JS}, year={1999}, month={Dec}, pages={187–193} }
 @article{guy_barnes_smith_breslin_1997, title={Antigenic characterization of a turkey coronavirus identified in poult enteritis- and mortality syndrome-affected turkeys}, volume={41}, ISSN={["1938-4351"]}, DOI={10.2307/1592148}, abstractNote={A turkey coronavirus (TCV [NC95]) was characterized by antigenic comparison with other avian and mammalian coronaviruses using immunofluorescence (FA) and immunoperoxidase (IP) procedures. Based on FA and IP procedures, TCV (NC95) was determined to be antigenically indistinguishable from turkey enteric (bluecomb) coronavirus (TECV). In addition, TCV (NC95) and TECV were found to be closely related to infectious bronchitis virus (IBV); a one-way antigenic relationship was demonstrated. Polyclonal antibodies specific for TECV and IBV reacted strongly against TCV (NC95), as determined by FA procedures. Monoclonal antibodies (MAbs) specific for IBV matrix protein (MAb 919) reacted strongly against TCV (NC95) and TECV as determined by FA and IP procedures; an IBV peplomer protein-specific MAb (MAb 94) did not recognize the two viruses. These studies suggest an identification of TCV (NC95) as a strain of TECV, and provide evidence of a close antigenic relationship between these viruses and IBV.}, number={3}, journal={AVIAN DISEASES}, author={Guy, JS and Barnes, HJ and Smith, LG and Breslin, J}, year={1997}, pages={583–590} }