Direct coating of culture medium from cells secreting classical swine fever virus E2 antigen on ELISA plates for detection of E2-specific antibodies

Cheng, Tian‐Lu; Pan, Chu-Hsiang; Chen, Chien Shu; Chuang, Kai‐Hsiang; Chuang, Chih Hung; Huang, Chien Chaio; Chu, Yu; Yang, Ya Chun; Chu, Pei Yu; Kao, Chien-Han; Hsieh, Yuan-Chin; Tian, Chen

doi:10.1016/j.tvjl.2015.02.007

Cited by 10 publications

(8 citation statements)

References 10 publications

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“…The authors suggested that as cELISA proved to be nearly as sensitive and specific as the VNT while being simpler and more rapid, it would be an adequate screening test for CDV or PDV cases [23]. The glycoprotein E2 exposed on the outer surface of CSFV is the major immuno-protective antigen of C-strain vaccine that is responsible for inducing neutralizing antibodies and eliciting protective immunity against CSFV [13][14][15][16][17][18][19][20]. Thus, it is not surprising that E2 protein has been successfully used to develop ELISAs to measure anti-CSFV antibody response in pigs after vaccination [13][14][15][16].…”

Section: Discussionmentioning

confidence: 99%

“…CSFV genome is comprised of a single large open reading frame (ORF) coding for a polyprotein encompassing all the viral proteins: four structural (C, E rns , E1, and E2) and eight nonstructural viral proteins (N pro , p7, NS2, NS3, NS4A, NS4B, NS5A, and NS5B) [11,12]. The envelope glycoprotein E2 is responsible for eliciting neutralizing antibodies which are protective against virulent CSF virus and is also the target antigen for development of CSF vaccines, molecular and serological tests [13][14][15][16]. High sequence variability has been found in E2 protein among CSFVs.…”

Section: Introductionmentioning

confidence: 99%

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A neutralizing monoclonal antibody-based competitive ELISA for classical swine fever C-strain post–vaccination monitoring

Wang

Madera

et al. 2020

BMC Vet Res

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Background: Virus neutralization test (VNT) is widely used for serological survey of classical swine fever (CSF) and efficacy evaluation of CSF vaccines. However, VNT is a time consuming procedure that requires cell culture and live virus manipulation. C-strain CSF vaccine is the most frequently used vaccine for CSF control and prevention. In this study, we presented a neutralizing monoclonal antibody (mAb) based competitive enzyme-linked immunosorbent assay (cELISA) with the emphasis on the replacement of VNT for C-strain post-vaccination monitoring. Results: One monoclonal antibody (6B211) which has potent neutralizing activity against C-strain was generated. A novel cELISA was established and optimized based on the strategy that 6B211 can compete with C-strain induced neutralizing antibodies in pig serum to bind capture antigen C-strain E2. By testing C-strain VNT negative pig sera (n = 445) and C-strain VNT positive pig sera (n = 70), the 6B211 based cELISA showed 100% sensitivity (95% confidence interval: 94.87 to 100%) and 100% specificity (95% confidence interval: 100 to 100%). The C-strain antibody can be tested in pigs as early as 7 days post vaccination with the cELISA. By testing pig sera (n = 139) in parallel, the cELISA showed excellent agreement (Kappa = 0.957) with VNT. The inhibition rate of serum samples in the cELISA is highly correlated with their titers in VNT (r 2 = 0.903, p < 0.001). In addition, intra-and inter-assays of the cELISA exhibited acceptable repeatability with low coefficient of variations (CVs). Conclusions: This novel cELISA demonstrated excellent agreement and high level correlation with VNT. It is a reliable tool for sero-monitoring of C-strain vaccination campaign because it is a rapid, simple, safe and cost effective assay that can be used to monitor vaccination-induced immune response at the population level.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A neutralizing monoclonal antibody-based competitive ELISA for classical swine fever C-strain post–vaccination monitoring

Wang

Madera

et al. 2020

BMC Vet Res

View full text Add to dashboard Cite

show abstract

“…In this study, we developed a neutralizing mAb based cELISA with the emphasis on the replacement of and specific as the VNT while being simpler and more rapid, it would be an adequate screening test for CDV or PDV cases [23]. The glycoprotein E2 exposed on the outer surface of CSFV is the major immuno-protective antigen of C-strain vaccine that is responsible for inducing neutralizing antibodies and eliciting protective immunity against CSFV [13][14][15][16][17][18][19][20]. Thus, it is not surprising that E2 protein has been successfully used to develop ELISAs to measure anti-CSFV antibody response in pigs after vaccination [13][14][15][16].…”

Section: Discussionmentioning

confidence: 99%

A neutralizing monoclonal antibody-based competitive ELISA for classical swine fever C-strain post–vaccination monitoring

Wang

Madera

et al. 2020

Preprint

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Background: Virus neutralization test (VNT) is widely used for serological survey of classical swine fever (CSF) and efficacy evaluation of CSF vaccines. However, VNT is a time consuming procedure that requires cell culture and live virus manipulation. C-strain CSF vaccine is the most frequently used vaccine for CSF control and prevention. In this study, we presented a neutralizing monoclonal antibody (mAb) based competitive enzyme-linked immunosorbent assay (cELISA) with the emphasis on the replacement of VNT for C-strain post–vaccination monitoring. Results: One monoclonal antibody (6B211) which has potent neutralizing activity against C-strain was generated. A novel cELISA was established and optimized based on the strategy that 6B211 can compete with C-strain induced neutralizing antibodies in pig serum to bind capture antigen C-strain E2. By testing C-strain VNT negative pig sera (n=445) and C-strain VNT positive pig sera (n=70), the 6B211 based cELSIA showed 100% sensitivity (95% confidence interval: 94.87 to 100%) and 100% specificity (95% confidence interval: 100 to 100%). The C-strain antibody can be detected in pigs as early as 7 days post vaccination with the cELISA. By testing pig sera (n=139) in parallel, the cELISA showed excellent agreement (Kappa=0.957) with VNT. The inhibition rate of serum samples in the cELISA is highly correlated with their titers in VNT (r 2 =0.903, p<0.001). In addition, intra- and inter-assays of the cELISA exhibited acceptable repeatability with low coefficient of variations (CVs). Conclusions: This novel cELISA demonstrated excellent agreement and high level correlation with VNT. It is a reliable tool for sero-monitoring of C-strain vaccination campaign because it is a rapid, simple, safe and cost effective assay that can be used to monitor vaccination-induced immune response at the population level.

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“…CSFV E2-specific antibodies, as important diagnostic targets (Ganges et al, 2008), have been commonly used for evaluating vaccination efficacy (Cheng et al, 2015), which is a key step in the control of CSF.…”

Section: Discussionmentioning

confidence: 99%

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

Zhang

et al. 2020

Transbound Emerg Dis

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The high performance of chemiluminescence immunoassays (CLIAs) in diagnosis has been gradually recognized in recent years, but their application in the diagnosis of classical swine fever (CSF) has not been reported. Here, a recombinant E2 (rE2) protein and a peroxidase‐conjugated monoclonal antibody (MAb G5) were used to develop a competition‐based chemiluminescence immunoassay (cCLIA) for rapid and accurate detection of E2‐specific antibodies in pig serum. To evaluate the feasibility of cCLIA in the diagnosis of CSF, we developed a competition‐based enzyme‐linked immunosorbent assay (cELISA) as a control. Under the optimum test conditions, cCLIA showed a higher signal‐to‐noise ratio than that of the control cELISA. The best signal‐to‐noise ratios of cCLIA and cELISA were 70 and 17, respectively. Then, the diagnostic performance of the two assays was compared by examining a panel of pig serum samples (n = 285) with a confirmed status, and cCLIA showed higher diagnostic sensitivity (Dn) and diagnostic specificity (Dp) values than those of cELISA. The Dn and Dp of cCLIA were 97.49% and 96.08%, respectively, and those of cELISA were 93.97% and 94.12%, respectively. Furthermore, cCLIA can provide results within 20 min, whereas the control cELISA requires at least 1 hr. According to these findings, the newly developed cCLIA has potential application in the diagnosis of CSF and offers an alternative approach for efficient and rapid detection of E2‐specific antibodies.

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Direct coating of culture medium from cells secreting classical swine fever virus E2 antigen on ELISA plates for detection of E2-specific antibodies

Cited by 10 publications

References 10 publications

A neutralizing monoclonal antibody-based competitive ELISA for classical swine fever C-strain post–vaccination monitoring

A neutralizing monoclonal antibody-based competitive ELISA for classical swine fever C-strain post–vaccination monitoring

A neutralizing monoclonal antibody-based competitive ELISA for classical swine fever C-strain post–vaccination monitoring

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

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