Immunodiagnosis of foot-and-mouth disease using mutated recombinant 3ABC polyprotein in a competitive ELISA

Sharma, Gaurav Kumar; Mohapatra, Jajati K.; Pandey, Laxmi; Mahajan, Sumit; Mathapati, Basavaraj; Sanyal, Aniket; Pattnaik, Bramhadev

doi:10.1016/j.jviromet.2012.05.029

Cited by 22 publications

(23 citation statements)

References 31 publications

(40 reference statements)

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“…The key protease active sites (amino acids 46 and 163) in the 3C protein must be modified to express the full-length 3ABC protein. Thus, a series of primers was designed according to the method described in a previous report (5). Briefly, the histidine at site 46 (CAC) was changed to tyrosine (TAC) using overlap extension PCR with two pairs of primers, 3ABC-F/3C-46-R and 3C-46-F/3ABC-R, for amplification (Table 3).…”

Section: Methodsmentioning

confidence: 99%

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Chemiluminescence Immunoassay for the Detection of Antibodies against the 2C and 3ABC Nonstructural Proteins Induced by Infecting Pigs with Foot-and-Mouth Disease Virus

Liu

Shao

Zhao

et al. 2017

Clin Vaccine Immunol

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The potential diagnostic value of chemiluminescence immunoassays (CLIAs) has been accepted in recent years, although their use for foot-and-mouth disease (FMD) diagnostics has not been reported. Full-length 3ABC and 2C proteins were expressed in bacteria and purified by affinity chromatography to develop a rapid and accurate approach to distinguish pigs infected with foot-and-mouth disease virus (FMDV) from vaccinated pigs. The recombinant proteins were then used as antigens to develop two CLIAs for the detection of antibodies against nonstructural viral proteins. The diagnostic performance of the two assays was compared by analyzing serum from pigs (naive pigs, n ϭ 63; vaccinated, uninfected pigs, n ϭ 532; naive, infected pigs, n ϭ 117) with a known infection status. The 3ABC-2C CLIA had a higher accuracy rate, with a diagnostic sensitivity of 100% and a diagnostic specificity of 96.5%, than the 3ABC CLIA, which had a diagnostic sensitivity of 95.7% and a diagnostic specificity of 96.0%. The results of the 3ABC-2C CLIA also had a high rate of concordance with those of two commercial FMDV enzyme-linked immunosorbent assay (ELISA) kits used to assess serum collected from 962 pigs in the field (96.2% and 97.8%, respectively). The 3ABC-2C CLIA detected infection in serum samples from infected pigs earlier than the commercial ELISA kits. In addition, the 3ABC-2C CLIA produced results within 15 min. On the basis of these findings, the 3ABC-2C CLIA could serve as the foundation for the development of penside FMD diagnostics and offers an alternative method to detect FMDV infections.

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Section: Methodsmentioning

confidence: 99%

“…been used to differentiate FMDV-infected animals from vaccinated animals (5). Among these methods, NSP-based ELISAs are the most widely used technique because they can detect seven different serotypes and discriminate infected from vaccinated animals (2,6).…”

mentioning

confidence: 99%

Chemiluminescence Immunoassay for the Detection of Antibodies against the 2C and 3ABC Nonstructural Proteins Induced by Infecting Pigs with Foot-and-Mouth Disease Virus

Liu

Shao

Zhao

et al. 2017

Clin Vaccine Immunol

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show abstract

“…The clone developed earlier [10] for expression of full length mutated 3ABC poly-protein in E. coli was used for the production of 3ABC antigen. Histidine tag fused 3ABC protein was purified under denaturating conditions by immobilized metal affinity chromatography using a commercial kit (Merck, Millipore, NJ, USA).…”

Section: Recombinant 3abc Antigen and Monoclonal Antibody Secreting Cmentioning

confidence: 99%

“…The scFv based ELISAs were standardized and validated by comparing the performance of the assays with the in-house 3ABC mAb based C-ELISA [10]. A panel of serum samples (n ¼ 2150) of known FMDV infection status was constituted for comparison and validation.…”

Section: Evaluation Of Scfv For Immunodiagnosis Of Fmdmentioning

confidence: 99%

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Production and characterization of single-chain antibody (scFv) against 3ABC non-structural protein in Escherichia coli for sero-diagnosis of Foot and Mouth Disease virus

et al. 2014

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New Approaches for Improvement of Diva Vaccines and Their Companion Diagnostic Tests for Foot-and-Mouth Disease and Bluetongue Disease

Prasad

Tiwari

Jain

et al. 2018

Advances in Animal Biotechnology and Its Applications

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Immunodiagnosis of foot-and-mouth disease using mutated recombinant 3ABC polyprotein in a competitive ELISA

Cited by 22 publications

References 31 publications

Chemiluminescence Immunoassay for the Detection of Antibodies against the 2C and 3ABC Nonstructural Proteins Induced by Infecting Pigs with Foot-and-Mouth Disease Virus

Chemiluminescence Immunoassay for the Detection of Antibodies against the 2C and 3ABC Nonstructural Proteins Induced by Infecting Pigs with Foot-and-Mouth Disease Virus

Production and characterization of single-chain antibody (scFv) against 3ABC non-structural protein in Escherichia coli for sero-diagnosis of Foot and Mouth Disease virus

New Approaches for Improvement of Diva Vaccines and Their Companion Diagnostic Tests for Foot-and-Mouth Disease and Bluetongue Disease

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