Identification of novel immunogenic proteins from Mycoplasma bovis and establishment of an indirect ELISA based on recombinant E1 beta subunit of the pyruvate dehydrogenase complex.

The pathogen Mycoplasma bovis (M. bovis) is a major cause of respiratory disease, mastitis, and arthritis in cattle. Screening the key immunogenic proteins and updating rapid diagnostic techniques are necessary to the prevention and control of M. bovis infection. In this study, 19 highly immunogenic...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Zhenhong Sun, Ping Fu, Kai Wei, Haiyan Zhang, Yuewei Zhang, Jian Xu, Fei Jiang, Xu Liu, Wei Xu, Wenxue Wu
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6fb9ca84242f464eadda84c3c6e140b8
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:6fb9ca84242f464eadda84c3c6e140b8
record_format	dspace
spelling	oai:doaj.org-article:6fb9ca84242f464eadda84c3c6e140b82021-11-18T08:33:09ZIdentification of novel immunogenic proteins from Mycoplasma bovis and establishment of an indirect ELISA based on recombinant E1 beta subunit of the pyruvate dehydrogenase complex.1932-620310.1371/journal.pone.0088328https://doaj.org/article/6fb9ca84242f464eadda84c3c6e140b82014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24520369/?tool=EBIhttps://doaj.org/toc/1932-6203The pathogen Mycoplasma bovis (M. bovis) is a major cause of respiratory disease, mastitis, and arthritis in cattle. Screening the key immunogenic proteins and updating rapid diagnostic techniques are necessary to the prevention and control of M. bovis infection. In this study, 19 highly immunogenic proteins from M. bovis strain PD were identified using 2-dimensional gel electrophoresis, immunoblotting and MALDI-TOF/TOF MS. Of these 19 proteins, pyruvate dehydrogenase E1 component beta subunit (PDHB) showed excellent immune reactivity and repeatability. PDHB was found to be conserved in different M. bovis isolates, as indicated by Western blot analysis. On the basis of these results, a rPDHB-based indirect ELISA (iELISA) was established for the detection of serum antibodies using prokaryotically expressed recombinant PDHB protein as the coating antigen. The specificity analysis result showed that rPDHB-based iELISA did not react with other pathogens assessed in our study except M. agalactiae (which infects sheep and goats). Moreover, 358 serum samples from several disease-affected cattle feedlots were tested using this iELISA system and a commercial kit, which gave positive rates of 50.8% and 39.9%, respectively. The estimated Kappa agreement coefficient between the two methods was 0.783. Notably, 39 positive serum samples that had been missed by the commercial kit were all found to be positive by Western blot analysis. The detection rate of rPDHB-based iELISA was significantly higher than that of the commercial kit at a serum dilution ratio of 1∶5120 to 1∶10,240 (P<0.05). Taken together, these results provide important information regarding the novel immunogenic proteins of M. bovis. The established rPDHB-based iELISA may be suitable for use as a new method of antibody detection in M. bovis.Zhenhong SunPing FuKai WeiHaiyan ZhangYuewei ZhangJian XuFei JiangXu LiuWei XuWenxue WuPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 2, p e88328 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Zhenhong Sun Ping Fu Kai Wei Haiyan Zhang Yuewei Zhang Jian Xu Fei Jiang Xu Liu Wei Xu Wenxue Wu Identification of novel immunogenic proteins from Mycoplasma bovis and establishment of an indirect ELISA based on recombinant E1 beta subunit of the pyruvate dehydrogenase complex.
description	The pathogen Mycoplasma bovis (M. bovis) is a major cause of respiratory disease, mastitis, and arthritis in cattle. Screening the key immunogenic proteins and updating rapid diagnostic techniques are necessary to the prevention and control of M. bovis infection. In this study, 19 highly immunogenic proteins from M. bovis strain PD were identified using 2-dimensional gel electrophoresis, immunoblotting and MALDI-TOF/TOF MS. Of these 19 proteins, pyruvate dehydrogenase E1 component beta subunit (PDHB) showed excellent immune reactivity and repeatability. PDHB was found to be conserved in different M. bovis isolates, as indicated by Western blot analysis. On the basis of these results, a rPDHB-based indirect ELISA (iELISA) was established for the detection of serum antibodies using prokaryotically expressed recombinant PDHB protein as the coating antigen. The specificity analysis result showed that rPDHB-based iELISA did not react with other pathogens assessed in our study except M. agalactiae (which infects sheep and goats). Moreover, 358 serum samples from several disease-affected cattle feedlots were tested using this iELISA system and a commercial kit, which gave positive rates of 50.8% and 39.9%, respectively. The estimated Kappa agreement coefficient between the two methods was 0.783. Notably, 39 positive serum samples that had been missed by the commercial kit were all found to be positive by Western blot analysis. The detection rate of rPDHB-based iELISA was significantly higher than that of the commercial kit at a serum dilution ratio of 1∶5120 to 1∶10,240 (P<0.05). Taken together, these results provide important information regarding the novel immunogenic proteins of M. bovis. The established rPDHB-based iELISA may be suitable for use as a new method of antibody detection in M. bovis.
format	article
author	Zhenhong Sun Ping Fu Kai Wei Haiyan Zhang Yuewei Zhang Jian Xu Fei Jiang Xu Liu Wei Xu Wenxue Wu
author_facet	Zhenhong Sun Ping Fu Kai Wei Haiyan Zhang Yuewei Zhang Jian Xu Fei Jiang Xu Liu Wei Xu Wenxue Wu
author_sort	Zhenhong Sun
title	Identification of novel immunogenic proteins from Mycoplasma bovis and establishment of an indirect ELISA based on recombinant E1 beta subunit of the pyruvate dehydrogenase complex.
title_short	Identification of novel immunogenic proteins from Mycoplasma bovis and establishment of an indirect ELISA based on recombinant E1 beta subunit of the pyruvate dehydrogenase complex.
title_full	Identification of novel immunogenic proteins from Mycoplasma bovis and establishment of an indirect ELISA based on recombinant E1 beta subunit of the pyruvate dehydrogenase complex.
title_fullStr	Identification of novel immunogenic proteins from Mycoplasma bovis and establishment of an indirect ELISA based on recombinant E1 beta subunit of the pyruvate dehydrogenase complex.
title_full_unstemmed	Identification of novel immunogenic proteins from Mycoplasma bovis and establishment of an indirect ELISA based on recombinant E1 beta subunit of the pyruvate dehydrogenase complex.
title_sort	identification of novel immunogenic proteins from mycoplasma bovis and establishment of an indirect elisa based on recombinant e1 beta subunit of the pyruvate dehydrogenase complex.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/6fb9ca84242f464eadda84c3c6e140b8
work_keys_str_mv	AT zhenhongsun identificationofnovelimmunogenicproteinsfrommycoplasmabovisandestablishmentofanindirectelisabasedonrecombinante1betasubunitofthepyruvatedehydrogenasecomplex AT pingfu identificationofnovelimmunogenicproteinsfrommycoplasmabovisandestablishmentofanindirectelisabasedonrecombinante1betasubunitofthepyruvatedehydrogenasecomplex AT kaiwei identificationofnovelimmunogenicproteinsfrommycoplasmabovisandestablishmentofanindirectelisabasedonrecombinante1betasubunitofthepyruvatedehydrogenasecomplex AT haiyanzhang identificationofnovelimmunogenicproteinsfrommycoplasmabovisandestablishmentofanindirectelisabasedonrecombinante1betasubunitofthepyruvatedehydrogenasecomplex AT yueweizhang identificationofnovelimmunogenicproteinsfrommycoplasmabovisandestablishmentofanindirectelisabasedonrecombinante1betasubunitofthepyruvatedehydrogenasecomplex AT jianxu identificationofnovelimmunogenicproteinsfrommycoplasmabovisandestablishmentofanindirectelisabasedonrecombinante1betasubunitofthepyruvatedehydrogenasecomplex AT feijiang identificationofnovelimmunogenicproteinsfrommycoplasmabovisandestablishmentofanindirectelisabasedonrecombinante1betasubunitofthepyruvatedehydrogenasecomplex AT xuliu identificationofnovelimmunogenicproteinsfrommycoplasmabovisandestablishmentofanindirectelisabasedonrecombinante1betasubunitofthepyruvatedehydrogenasecomplex AT weixu identificationofnovelimmunogenicproteinsfrommycoplasmabovisandestablishmentofanindirectelisabasedonrecombinante1betasubunitofthepyruvatedehydrogenasecomplex AT wenxuewu identificationofnovelimmunogenicproteinsfrommycoplasmabovisandestablishmentofanindirectelisabasedonrecombinante1betasubunitofthepyruvatedehydrogenasecomplex
_version_	1718421628081143808

Identification of novel immunogenic proteins from Mycoplasma bovis and establishment of an indirect ELISA based on recombinant E1 beta subunit of the pyruvate dehydrogenase complex.

Ejemplares similares