Elevated Mhp462 antibody induced by natural infection but not in vitro culture of Mycoplasma hyopneumoniae

Ning, Yaru; Zhou, Yuqing; Wang, Zhaodi; Wen, Yukang; Xu, Zuobo; Tian, Yaqin; Yang, Mei; Wang, Xudong; Yang, Yujiao; Ding, Honglei

doi:10.1016/j.heliyon.2020.e04832

Cited by 1 publication

(1 citation statement)

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“…Based on the characteristics of the Mhp366 protein, we developed two ELISAs, one for screening serological immunodominant protein antigens [ 12 ] and another for further screening the discriminative immunodominant proteins that can distinguish between anti- M. hyopneumoniae IgG in inactivated bacterin-induced hyperimmune sera and convalescent sera resulting from natural M. hyopneumoniae infection [ 13 ]. The studies identified 15 serological immunodominant proteins and 1 discriminative serological immunodominant protein, Mhp462 [ 14 ]. These data suggest that a Mhp366-based ELISA has potential to be used as a diagnostic tool to detect natural M. hyopneumoniae infection.…”

Section: Introductionmentioning

confidence: 99%

Development of an indirect ELISA for detection of anti-Mycoplasma hyopneumoniae IgG in naturally infected pathogen-induced convalescent sera

Tian

Wen

et al. 2021

BMC Vet Res

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Background Immunization of pigs with an inactivated Mycoplasma hyopneumoniae vaccine (bacterin) generates hyperimmune serum that contains high concentrations of anti-M. hyopneumoniae IgG. Commercially available IgG-ELISA kits cannot distinguish between anti-M. hyopneumoniae IgG in inactivated bacterin-induced hyperimmune sera and convalescent sera resulting from natural M. hyopneumoniae infection. Establishment of an ELISA to detect anti-M. hyopneumoniae IgG in convalescent sera will facilitate the evaluation of the M. hyopneumoniae status of pig farms. Results In this study, we expressed and purified recombinant Mhp366-N protein, which contains an epitope recognized by M. hyopneumoniae convalescent sera but not hyperimmune sera, for use as a coating antigen. For the M. hyopneumoniae convalescent serum IgG-ELISA, the optimal antigen concentration, blocking buffer, blocking time, dilution of serum, incubation time with serum, secondary antibody dilution, secondary antibody incubation time and colorimetric reaction time were 0.25 µg/mL, 2.5 % skim milk, 1 h, 1:500, 0.5 h, 1:10,000, 1 h and 15 min, respectively. Validation of the M. hyopneumoniae convalescent serum IgG-ELISA showed a cut-off value of 0.323, the intra-assay CV ranged from 3.27 to 7.26 %, the inter-assay CV ranged from 3.46 to 5.93 %, and the assay was able to differentiate convalescent sera from antibodies to 7 other porcine respiratory pathogens. The convalescent serum IgG-ELISA detected no anti-M. hyopneumoniae IgG in hyperimmune serum samples while a commercial IgG-ELISA identified 95/145 of these sera as positive. The accuracy of the M. hyopneumoniae convalescent serum IgG-ELISA was comparable to the sIgA-ELISA but better than the commercial IgG-ELISA. Conclusions The convalescent serum IgG-ELISA is a reproducible, sensitive, and specific indirect ELISA to detect anti-M. hyopneumoniae IgG in naturally infected pathogen-induced convalescent sera. This ELISA could be used to carry out large scale surveillance of M. hyopneumoniae infection in pig farms regardless of vaccination status.

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Section: Introductionmentioning

confidence: 99%