{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,18]],"date-time":"2026-04-18T08:55:45Z","timestamp":1776502545864,"version":"3.51.2"},"reference-count":24,"publisher":"Springer Science and Business Media LLC","issue":"1","content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Virol J"],"published-print":{"date-parts":[[2010,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background<\/jats:title>\n            <jats:p>The duck plague virus (DPV) UL46 protein (VP11\/12) is a 739-amino acid tegument protein encoded by the <jats:italic>UL46<\/jats:italic> gene. We analyzed the amino acid sequence of UL46 using bioinformatics tools and defined the main antigenic domains to be between nucleotides 700-2,220 in the <jats:italic>UL46<\/jats:italic> sequence. This region was designated UL46M. The DPV <jats:italic>UL46<\/jats:italic> and <jats:italic>UL46M<\/jats:italic> genes were both expressed in <jats:italic>Escherichia coli<\/jats:italic> Rosetta (DE3) induced by isopropy1-\u03b2-<jats:sc>D<\/jats:sc>-thiogalactopyranoside (IPTG) following polymerase chain reaction (PCR) amplification and subcloning into the prokaryotic expression vector pET32a(+). The recombinant proteins were purified using a Ni-NTA spin column and used to generate the polyclonal antibody against UL46 and UL46M in New Zealand white rabbits. The titer was then tested using enzyme-linked immunosorbent assay (ELISA) and agar diffusion reaction, and the specificity was tested by western blot analysis. Subsequently, we established Dot-ELISA using the polyclonal antibody and applied it to DPV detection.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>In our study, the DPV UL46M fusion protein, with a relative molecular mass of 79 kDa, was expressed in <jats:italic>E. coli<\/jats:italic> Rosetta (DE3). Expression of the full <jats:italic>UL46<\/jats:italic> gene failed, which was consistent with the results from the bioinformatic analysis. The expressed product was directly purified using Ni-NTA spin column to prepare the polyclonal antibody against UL46M. The titer of the anti-UL46M antisera was over 1:819,200 as determined by ELISA and 1:8 by agar diffusion reaction. Dot-ELISA was used to detect DPV using a 1:60 dilution of anti-UL46M IgG and a 1:5,000 dilution of horseradish peroxidase (HRP)-labeled goat anti-rabbit IgG.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions<\/jats:title>\n            <jats:p>The anti-UL46M polyclonal antibody reported here specifically identifies DPV, and therefore, it is a promising diagnostic tool for DPV detection in animals. UL46M and the anti-UL46M antibody can be used for further clinical examination and research of DPV.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1743-422x-7-83","type":"journal-article","created":{"date-parts":[[2010,4,29]],"date-time":"2010-04-29T18:14:21Z","timestamp":1272564861000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Polyclonal antibody against the DPV UL46M protein can be a diagnostic candidate"],"prefix":"10.1186","volume":"7","author":[{"given":"Liting","family":"Lu","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anchun","family":"Cheng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mingshu","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jinfeng","family":"Jiang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dekang","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Renyong","family":"Jia","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qihui","family":"Luo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fei","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhengli","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoyue","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jinlong","family":"Yang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2010,4,29]]},"reference":[{"key":"828_CR1","first-page":"354","volume-title":"Diseases of Poultry","author":"TS Sandhu","year":"2003","unstructured":"Sandhu TS, Shawky SA: Duck virus enteritis (duck plague). In Diseases of Poultry. Edited by: Saif YM, Barnes HJ, Glisson, JR. Ames: Iowa State University Press; 2003:354-363."},{"key":"828_CR2","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1159\/000150328","volume":"36","author":"R Gardner","year":"1993","unstructured":"Gardner R, Wilkerson J, Johnson JC: Molecular characterization of the DNA of Anatid herpesvirus 1. Intervirology 1993, 36: 99-112.","journal-title":"Intervirology"},{"key":"828_CR3","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1080\/03079459008418673","volume":"19","author":"EF Kaleta","year":"1990","unstructured":"Kaleta EF: Herpesviruses of birds-a review. Avian Pathol 1990, 19: 193-211. 10.1080\/03079459008418673","journal-title":"Avian Pathol"},{"key":"828_CR4","doi-asserted-by":"publisher","first-page":"308","DOI":"10.1637\/0005-2086(2002)046[0308:LSAROD]2.0.CO;2","volume":"46","author":"S Shawky","year":"2002","unstructured":"Shawky S, Schta KA: Latency sites and reactivation of duck enteritis virus. Avian Dis 2002, 46: 308-313. 10.1637\/0005-2086(2002)046[0308:LSAROD]2.0.CO;2","journal-title":"Avian Dis"},{"key":"828_CR5","volume-title":"Virus Taxonomy: Eighth Report of the International Committee on Taxonomy of Viruses","author":"CM Fauquet","year":"2005","unstructured":"Fauquet CM, Fauquet C, Mayo MA, Maniloff J: Virus Taxonomy: Eighth Report of the International Committee on Taxonomy of Viruses. San Diego, Elsevier; 2005."},{"key":"828_CR6","first-page":"231","volume":"25","author":"AC Cheng","year":"2005","unstructured":"Cheng AC, Wang MS, Liu F, Song Y, Yuan GP, Han XY, Xu C, Liao YH, Wen M, Zhou WG, Jia RY: Distribution and excretion of duck plague virus attenuated Chv strain in vaccinated ducklings. Chinese Journal of Veterinary Science 2005, 25: 231-233. (in Chinese)","journal-title":"Chinese Journal of Veterinary Science"},{"key":"828_CR7","doi-asserted-by":"publisher","first-page":"338","DOI":"10.1637\/8120-100207-ResNote.1","volume":"52","author":"XF Qi","year":"2008","unstructured":"Qi XF, Yang XY, Cheng AC, Wang MS, Zhu DK, Jia RY: Quantitative analysis of virulent duck enteritis virus loads in experimentally infected ducklings. Avian Dis 2008, 52: 338-344. 10.1637\/8120-100207-ResNote.1","journal-title":"Avian Dis"},{"key":"828_CR8","doi-asserted-by":"publisher","first-page":"50","DOI":"10.1637\/7237-071004R","volume":"49","author":"GP Yuan","year":"2005","unstructured":"Yuan GP, Cheng AC, Wang MS, Liu F, Han XY, Liao YH, Xu C: Electron microscopic studies of the morphogenesis of duck enteritis virus. Avian Dis 2005, 49: 50-55. 10.1637\/7237-071004R","journal-title":"Avian Dis"},{"key":"828_CR9","first-page":"202","volume":"23","author":"RY Jia","year":"2007","unstructured":"Jia RY, Cheng AC, Wang MS, Guo YF, Wen M, Xu C, Yuan GP, Zhou WG, Zhou Y, Chen XY: Studies of ultrastructure of duck enteritis virus CHv virulent strain. Chinese Journal of Virology 2007, 23: 202-205. (in Chinese)","journal-title":"Chinese Journal of Virology"},{"key":"828_CR10","first-page":"486","volume":"36","author":"GP Yuan","year":"2005","unstructured":"Yuan GP, Cheng AC, Wang MS, Zhou Y, Liu F, Han XY, Guo YF, Liao YH, Xu C, Wen M, Jia RY, Zhou WG, Chen XY: Electron microscopic studies on morphology and morphogenesis of duck enteritis virus in ducks infected with DEV virulent strain experimentally. Acta Veterinaria et Zootechnica Sinica 2005, 36: 486-491. (in Chinese)","journal-title":"Acta Veterinaria et Zootechnica Sinica"},{"key":"828_CR11","doi-asserted-by":"publisher","first-page":"266","DOI":"10.2307\/1592539","volume":"44","author":"WR Hansen","year":"2000","unstructured":"Hansen WR, Nashold SW, Docherty DE, Brown SE, Knudson DL: Diagnosis of duck plague in waterfowl by polymerase chain reaction. Avian Dis 2000, 44: 266-274. 10.2307\/1592539","journal-title":"Avian Dis"},{"key":"828_CR12","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1128\/jvi.14.1.8-19.1974","volume":"14","author":"RW Honess","year":"1974","unstructured":"Honess RW, Roizman B: Regulation of herpesvirus macromolecular synthesis.I. Cascade regulation of the synthesis of there groups of viral proteins. Journal of Virology 1974, 14: 8.","journal-title":"Journal of Virology"},{"key":"828_CR13","doi-asserted-by":"publisher","first-page":"1531","DOI":"10.1099\/0022-1317-69-7-1531","volume":"69","author":"DJ Mcgeoch","year":"1988","unstructured":"Mcgeoch DJ, Dalrymple MA, Davison AJ, Dolan A, Frame MC, McNab D, Perry LJ, Scott JE, Taylor P: The complete DNA sequence of the unique long region in the genome of herpes simplex virus type 1. J Gen Virol 1988, 69: 1531-1574. 10.1099\/0022-1317-69-7-1531","journal-title":"J Gen Virol"},{"key":"828_CR14","first-page":"125","volume":"154","author":"MS Chee","year":"1990","unstructured":"Chee MS, Bankier AT, Beck S, Bohni R, Brown CM, Cerny R, Horsnell T, Hutchison CA, Kouzarides T, Martignetti JA: Analysis of the protein coding content of the sequence of human cytomegalovirus strain AD169. Curr Top Microbiol Immunol 1990, 154: 125-169.","journal-title":"Curr Top Microbiol Immunol"},{"key":"828_CR15","doi-asserted-by":"publisher","first-page":"207","DOI":"10.1038\/310207a0","volume":"310","author":"R Baer","year":"1984","unstructured":"Baer R, Bankier AT, Biggin MD, Deininger PL, Farrell PJ, Gibson TJ, Hatfull G, Hudson GS, Satchwell SC, Seguin C, Tuffnell PS, Barrell BG: DNA sequence and expression of the B95-8 Epstein-Barr virus genome. Nature 1984, 310: 207-211. 10.1038\/310207a0","journal-title":"Nature"},{"key":"828_CR16","doi-asserted-by":"publisher","first-page":"12891","DOI":"10.1128\/JVI.77.23.12891-12900.2003","volume":"77","author":"W Fuchs","year":"2003","unstructured":"Fuchs W, Granzow H, Mettenleiter TC: A Pseudorabies Virus Recombinant Simultaneously Lacking the Major Tegument Proteins Encoded by the UL46, UL47, UL48, and UL49 Genes Is Viable in Cultured Cells. J Virol 2003, 77: 12891-12900. 10.1128\/JVI.77.23.12891-12900.2003","journal-title":"J Virol"},{"key":"828_CR17","doi-asserted-by":"crossref","first-page":"1482","DOI":"10.1128\/jvi.67.3.1482-1492.1993","volume":"67","author":"YZ Zhang","year":"1993","unstructured":"Zhang YZ, Mcknight JL: Herpes simplex virus type 1 UL46 and UL47 deletion mutants lack VP11 and VP12 or VP13 and VP14, respectively, and exhibit altered viral thymidine kinase expression. J Virol 1993, 67: 1482-1492.","journal-title":"J Virol"},{"key":"828_CR18","doi-asserted-by":"publisher","first-page":"2149","DOI":"10.1007\/s007050070045","volume":"14","author":"K Kato","year":"2000","unstructured":"Kato K, Daikoku T, Goshima F, Kume H, Yamaki K, Nishiyama Y: Synthesis, subcellular localization and VP16 interaction of the herpes simplex virus type 2 UL46 gene product. Arch Virol 2000, 14: 2149-2162. 10.1007\/s007050070045","journal-title":"Arch Virol"},{"key":"828_CR19","first-page":"27","volume-title":"Molecular Cloning: A Laboratory Manual","author":"J Sambrook","year":"2003","unstructured":"Sambrook J, Russell DW: Molecular Cloning: A Laboratory Manual. 3rd edition. Edited by: Huang Pei-tang, Wang Jia-xi, Zhu Hou-chu, et al. Beijing: Science Press; 2003:27-30. 96-99. (in Chinese)","edition":"3"},{"key":"828_CR20","first-page":"948","volume":"16","author":"AC Cheng","year":"2006","unstructured":"Cheng AC, Wang MS, Wen M, Zhou WG, Guo YF, Jia RY, Xu C, Yuan GP, Liu YC: Construction of duck enteritis virus gene libraries and discovery, cloning and identification of viral nucleocapsid protein gene. High Technol Lett 2006, 16: 948-953.","journal-title":"High Technol Lett"},{"key":"828_CR21","doi-asserted-by":"publisher","first-page":"141","DOI":"10.1159\/000221833","volume":"52","author":"MS Cai","year":"2009","unstructured":"Cai MS, Cheng AC, Wang MS, Zhao LC, Zhu DK, Luo QH, Liu F, Chen XY: His-Tagged UL46 protein of duck plague virus: expression, purification and production of polyclonal antibody. intervirology 2009, 52: 141-151. 10.1159\/000221833","journal-title":"intervirology"},{"key":"828_CR22","doi-asserted-by":"publisher","first-page":"248","DOI":"10.1016\/0003-2697(76)90527-3","volume":"72","author":"M Bradford","year":"1976","unstructured":"Bradford M: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 1976, 72: 248-254. 10.1016\/0003-2697(76)90527-3","journal-title":"Analytical Biochemistry"},{"key":"828_CR23","doi-asserted-by":"publisher","first-page":"157","DOI":"10.1038\/sj.cr.7290122","volume":"12","author":"YX Hu","year":"2002","unstructured":"Hu YX, Guo JY, Shen L, Chen Y, Zhang ZC, Zhang YL: Get effective polyclonal antisera in one month. Cell res 2002, 12: 157-160. 10.1038\/sj.cr.7290122","journal-title":"Cell res"},{"key":"828_CR24","unstructured":"Walker JM: Purification of IgG by precipitation with sodium sulfate or ammonium sulphate. In The protein protocols handbook. second edition. Humana press; 101385\/1-59259-169-8: 983-984 (copyright 2002)"}],"container-title":["Virology Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/1743-422X-7-83.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,9,1]],"date-time":"2021-09-01T12:18:24Z","timestamp":1630498704000},"score":1,"resource":{"primary":{"URL":"https:\/\/virologyj.biomedcentral.com\/articles\/10.1186\/1743-422X-7-83"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2010,4,29]]},"references-count":24,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2010,12]]}},"alternative-id":["828"],"URL":"https:\/\/doi.org\/10.1186\/1743-422x-7-83","relation":{},"ISSN":["1743-422X"],"issn-type":[{"value":"1743-422X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2010,4,29]]},"assertion":[{"value":"3 February 2010","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"29 April 2010","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"29 April 2010","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}}],"article-number":"83"}}