A chemiluminescence immunoassay for rapid detection of classical swine fever virus E2 antibodies in pig serum samples

Ma, Zhongyuan; Lv, Jing; Zhang, Zhongwang; Zhao, Ye; Pan, Li; Zhang, Yongguang

doi:10.1111/tbed.13557

Cited by 2 publications

(1 citation statement)

References 20 publications

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“…CSFV E2 positive control and negative control sera were prepared by our group [ 19 ]. A standard positive control (P) and negative control (N) were prepared as internal controls.…”

Section: Methodsmentioning

confidence: 99%

Development and application of classical swine fever virus monoclonal antibodies derived from single B cells

Ma,

Zhao,

et al. 2023

Vet Res

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Vaccination with E2 subunit vaccines is currently the main measure to control classical swine fever virus (CSFV), which is an endemic disease, and detection of antibodies against CSFV E2 is the most effective way to evaluate herd immunity. In the present study, the E2 protein was expressed by a baculovirus expression system, and two monoclonal antibodies (mAbs), namely, 3A9 and 4F7, were successfully produced using techniques for the isolation of single B cells from splenocytes from mice immunized with the E2 protein. Moreover, two linear B-cell epitopes, 25GLTTTWKEYSHDLQL39 and 259GNTTVKVHASDERGP273, reactive to 3A9 and 4F7, respectively, were identified using epitope mapping of the E2 protein. In addition, the diagnostic performance of the two mAbs was evaluated using blocking enzyme-linked immunosorbent assay (bELISA), and the results showed that the two mAbs had high diagnostic specificity (96.08%, 94.38%) and diagnostic sensitivity (97.49%, 95.97%). Together, these findings identify two ideal candidate peptides and matching mAbs for a new method of CSFV diagnosis, which will contribute to the control and eradication of classical swine fever.

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“…CSFV E2 positive control and negative control sera were prepared by our group [ 19 ]. A standard positive control (P) and negative control (N) were prepared as internal controls.…”

Section: Methodsmentioning

confidence: 99%

Development and application of classical swine fever virus monoclonal antibodies derived from single B cells

Ma,

Zhao,

et al. 2023

Vet Res

View full text Add to dashboard Cite

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Development of a fully automated chemiluminescent immunoassay for the quantitative and qualitative detection of antibodies against African swine fever virus p72

Wang,

Li,

Zeng

et al. 2024

Microbiol Spectr

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African swine fever (ASF), caused by ASF virus (ASFV), is a highly infectious and severe hemorrhagic disease of pigs that causes major economic losses. Currently, no commercial vaccine is available and prevention and control of ASF relies mainly on early diagnosis. Here, a novel automated double antigen sandwich chemiluminescent immunoassay (DAgS-aCLIA) was developed to detect antibodies against ASFV p72 (p72-Ab). For this purpose, recombinant p72 trimer was produced, coupled to magnetic particles as carriers and labeled with acridinium ester as a signal trace. Finally, p72-Ab can be sensitively and rapidly measured on an automated chemiluminescent instrument. For quantitative analysis, a calibration curve was established with a laudable linearity range of 0.21 to 212.0 ng/mL (R 2 = 0.9910) and a lower detection limit of 0.15 ng/mL. For qualitative analysis, a cut-off value was set at 1.50 ng/mL with a diagnostic sensitivity of 100.00% and specificity of 98.33%. Furthermore, antibody response to an ASF gene-deleted vaccine candidate can be accurately quantified using this DAgS-aCLIA, as evidenced by early seroconversion as early as 7 days post-immunization and high antibody levels. Compared with available enzyme-linked immunosorbent assays, this DAgS-aCLIA demonstrated a wider linearity range of 4 to 16-fold, and excellent analytical sensitivity and agreement of over 95.60%. In conclusion, our proposed DAgS-aCLIA would be an effective tool to support ASF epidemiological surveillance. IMPORTANCE African swine fever virus (ASFV) is highly contagious in wild boar and domestic pigs. There is currently no vaccine available for ASF, so serological testing is an important diagnostic tool. Traditional enzyme-linked immunosorbent assays provide only qualitative results and are time and resource consuming. This study will develop an automated chemiluminescent immunoassay (CLIA) that can quantitatively and qualitatively detect antibodies to ASFV p72, greatly reducing detection time and labour-intensive operation, and improving detection sensitivity and linearity range. This novel CLIA would serve as a reliable and convenient tool for ASF pandemic surveillance and vaccine development.

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A chemiluminescence immunoassay for rapid detection of classical swine fever virus E2 antibodies in pig serum samples

Cited by 2 publications

References 20 publications

Development and application of classical swine fever virus monoclonal antibodies derived from single B cells

Development and application of classical swine fever virus monoclonal antibodies derived from single B cells

Development of a fully automated chemiluminescent immunoassay for the quantitative and qualitative detection of antibodies against African swine fever virus p72

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