About: Development of luciferase-linked antibody capture assay based on luciferase immunoprecipitation systems for antibody detection of porcine reproductive and respiratory syndrome virus

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About: Development of luciferase-linked antibody capture assay based on luciferase immunoprecipitation systems for antibody detection of porcine reproductive and respiratory syndrome virus Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : wasabi.inria.fr associated with source document(s)

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type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Development of luciferase-linked antibody capture assay based on luciferase immunoprecipitation systems for antibody detection of porcine reproductive and respiratory syndrome virus
Creator	Wang, Gang Nan, Yuchen Wu, Chunyan Cai, Xuehui Liu, Yonggang Zhou, En-Min Zhao, Bao Li, Jie Yang, Di Zhao, Yongpan
Source	Medline; PMC
abstract	BACKGROUND: Early detection of porcine reproductive and respiratory syndrome virus (PRRSV) infection of swine is necessary to control this devastating disease. By monitoring host serum antibodies to viral antigens, early virus detection within herds is feasible. In this study, recombinant antigens were generated using recombinant DNA techniques to fuse PRRSV structural protein (N) or nonstructural protein 1α (nsp1α) with the Rellina luciferase gene. Next, fused genes were cloned into plasmids and transfected into HEK-293 T cells for transient expression. Upon co-incubation of lysates with pig sera, antigen-antibody complexes formed that bound to Protein-G coated onto microplates. By further measurement of luminance value, a modified form of Luciferase Immunoprecipitation Systems, namely luciferase-linked antibody capture assay (LACA) was developed for detection of PRRSV-specific antibodies. RESULTS: Known anti-PRRSV antibody-positive or -negative serum samples (125 and 122 samples, respectively) were used to validate the LACA and compared it with IDEXX PRRS ×3 ELISA. Based on the result, N-Rluc and nsp1α-Rluc LACA results were 95.3 and 94.4% in agreement with IDEXX ELISA, suggested a similar specificity of LACA to IDEXX ELISA. Moreover, when both LACA and IDEXX ELISA were used to evaluate sequential serum samples obtained from PRRSV experimentally infected pigs, the PRRSV-specific antibody response was detectable as early as 3 days post-inoculation (dpi) using N-Rluc LACA, but undetectable until 7 dpi using IDEXX ELISA, suggesting an improved sensitivity of LACA. Meanwhile, antibodies specific for nsp1α were detected at higher levels overall, but were undetectable until 10 dpi. Furthermore,. Notably, one IDEXX ELISA positive result was not confirmed by LACA or IFA and was thus considered a false-positive result. CONCLUSION: The LACA exhibited similar specificity but improved sensitivity to that of the commercial IDEXX PRRS ×3 ELISA kit for detection of PRRSV-specific antibodies in pig serum. Importantly, LACA could be adapted for detecting antibodies against other PRRSV targets, such as nsp1α, to achieve earlier detection of PRRSV infection.
has issue date	2018-11-16(xsd:dateTime)
bibo:doi	10.1186/s12896-018-0483-5
bibo:pmid	30445953
has license	cc-by
sha1sum (hex)	c7bf679ec25305b15089e89885f393de974be10d
schema:url	https://doi.org/10.1186/s12896-018-0483-5
resource representing a document's title	Development of luciferase-linked antibody capture assay based on luciferase immunoprecipitation systems for antibody detection of porcine reproductive and respiratory syndrome virus
has PubMed Central identifier	PMC6240198
has PubMed identifier	30445953
schema:publication	BMC Biotechnol
resource representing a document's body	covid:c7bf679ec25305b15089e89885f393de974be10d#body_text
is schema:about of	named entity 'protein' named entity 'detection' named entity 'antibody' named entity 'measurement' named entity 'result' named entity 'detection' named entity 'pig' named entity 'fuse' named entity 'porcine' named entity 'antibody' covid:arg/c7bf679ec25305b15089e89885f393de974be10d named entity 'developed' named entity 'cloned' named entity 'specificity' named entity 'bound' named entity 'recombinant DNA' named entity 'fused' named entity 'Immunoprecipitation' named entity 'study' named entity 'levels' named entity 'Next' named entity 'pig' named entity 'herds' named entity 'goat' named entity 'antibodies' named entity 'cell lysate' named entity 'P-value' named entity 'Western blot' named entity 'antibodies' named entity 'PRRS' named entity 'undetectable' named entity 'suggested' named entity 'monitoring' named entity 'antibodies' named entity 'modified' named entity 'respiratory' named entity 'LACA' named entity 'T cells' named entity 'luciferase' named entity 'gene' named entity 'LACA' named entity 'transfected' named entity 'infection' named entity 'LACA' named entity 'Immunoprecipitation' named entity 'assay' named entity 'antibody' named entity 'LACA' named entity 'antigen-antibody complexes' named entity 'antibody' named entity 'LACA' named entity 'plasmids' named entity 'false-positive result' named entity 'immunoprecipitation' named entity 'antibody' named entity 'Corning, NY' named entity 'immunochromatographic' named entity 'false negative results' named entity 'antibody' named entity 'clone' named entity 'ad libitum' named entity 'Sigma-Aldrich' named entity 'porcine circovirus' named entity 'coding sequence' named entity 'PRRSV' named entity 'cell lysis' named entity 'Protein' named entity 'Renilla luciferase' named entity 'supernatants' named entity 'mammalian cells' named entity 'positive controls' named entity 'enzymatic activity'

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