Establishment of an antibody avidity test to differentiate vaccinated cattle from those naturally infected with Mycoplasma bovis

Han, Xing; Khan, Farhan Anwar; Zhu, Xifang; Zhang, Rui; Mustafa, Riaz; Hu, Changmin; Chen, Yingyu; Chen, Huanchun

doi:10.1016/j.tvjl.2014.10.032

Cited by 22 publications

(31 citation statements)

References 22 publications

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“…Thirty-six clinically healthy 5-6 month old local calves were confirmed to be free of M. bovis as described previously [49]. These animals were randomly allocated among 3 groups.…”

Section: Methodsmentioning

confidence: 99%

“…The animals were euthanized and necropsied 40 days after infection/vaccination for gross lesion evaluation using a previously described scoring system [3]. The antibody titers of serum samples were measured by iELISA as described previously [49]. Serum samples collected on day 0 before infection or immunization were used as negative controls.…”

Section: Methodsmentioning

confidence: 99%

“…Disease was diagnosed as described previously [49]. All serum samples were stored at −80°C for further analysis.…”

Section: Methodsmentioning

confidence: 99%

“…iELISA conditions were optimized for M. bovis WCPs and each recombinant protein as described elsewhere [49]. In brief, 96-well microtitre plates were coated overnight at 4°C with 125 ng of WCPs and 250 ng of each purified recombinant protein diluted in 100 μL sodium carbonate buffer (pH 9.6) and washed with PBS containing 0.05% Tween 20 (PBST).…”

Section: Methodsmentioning

confidence: 99%

“…An iELISA was established as described previously [49]. Briefly, 96-well ELISA plates were coated with rMbovP579 serially diluted two-fold from a concentration of 2 μg to 0.97 ng/well and incubated at 4°C overnight.…”

Section: Methodsmentioning

confidence: 99%

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Immunoproteomic identification of MbovP579, a promising diagnostic biomarker for serological detection of Mycoplasma bovis infection

et al. 2016

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A lack of knowledge regarding the antigenic properties of Mycoplasma bovis proteins prevents the effective control of bovine infections using immunological approaches. In this study, we detected and characterized a specific and sensitive M. bovis diagnostic biomarker. After M. bovis total proteins and membrane fractions were separated with two dimensional gel electrophoresis, proteins reacting with antiserawere detected using MALDI-TOF MS. Thirty-nine proteins were identified, 32 of which were previously unreported. Among them, immunoinformatics predicted eight antigens, encoded by Mbov_0106, 0116, 0126, 0212, 0275, 0579, 0739, and 0789, to have high immunological value. These genes were expressed in E. coli after mutagenesis of UGA to UGG using overlap extension PCR. A lipoprotein, MbovP579, encoded by a functionally unknown gene, was a sensitive and specific antigen for detection of antibodies in sera from both M. bovis-infected and vaccinated cattle. The specificity of MbovP579 was confirmed by its lack of cross-reactivity with other mycoplasmas, including Mycoplasma agalactiae. An iELISA based on rMbovP579 detected seroconversion 7 days post-infection (dpi). The ELISA had sensitivity of 90.2% (95% CI: 83.7%, 94.3%) and a specificity of 97.8% (95% CI: 88.7%, 99.6%) with clinical samples. Additional comparative studies showed that both diagnostic and analytic sensitivities of the ELISA were higher than those of a commercially available kit (p<0.01). We have thus detected and characterized the novel antigen, MbovP579, and established an rMbovP579-based ELISA as a highly sensitive and specific method for the early diagnosis of M. bovis infection.

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“…Thirty-six clinically healthy 5-6 month old local calves were confirmed to be free of M. bovis as described previously [49]. These animals were randomly allocated among 3 groups.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Disease was diagnosed as described previously [49]. All serum samples were stored at −80°C for further analysis.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations