Development of an indirect ELISA assay for the detection of IgG antibodies against the ORF1 of Torque teno sus viruses 1 and 2 in conventional pigs

Nieto, D.; Martínez-Guinó, Laura; Jiménez-Melsió, Alexandra; Segalés, Joaquím; Kekarainen, Tuija

doi:10.1016/j.vetmic.2015.08.023

Cited by 5 publications

(2 citation statements)

References 38 publications

(54 reference statements)

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“…A conformational change in the precise site of dominant epitopes can have a relevant impact in immune function, particularly on immunoglobulin production. It has been found that several IgGs are able to bind to ORF1 (Singh & Ramamoorthy, ; Jarasova & Celer, ; Liu et al., ; Giménez‐Lirola et al., ; Nieto, Martínez‐Guinó, Jiménez‐Melsio, Segalés, & Kekarainen, ) and ORF2 products (Jiménez‐Melsió et al., ) from TTSuV1a and TTSuV1b.…”

Section: Discussionmentioning

confidence: 99%

Molecular characterization of the ORF2 of Torque teno sus virus 1a and Torque teno sus virus 1b detected in cases of postweaning multisystemic wasting syndrome in Mexico

Vargas-Ruiz¹,

García-Camacho²,

Álvarez³

et al. 2018

Transbound Emerg Dis

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Worldwide Torque teno sus virus (TTSuV, genus Iotatorquevirus) species have been regarded as possible agents associated with porcine circovirus-associated disease. Iotatorquevirus species possess high genomic variability, suggesting that diverse genotypes are widely geographically distributed. In this study, we validated the genomic variability of Iotaroquevirus species in pigs with postweaned multisystemic wasting syndrome. Genomic DNA from nine TTSuV1a-positive tissues and 15 TTSuV1b-positive tissues was used to amplify the complete ORF2 of each species by nested PCR to perform a molecular characterization. It was found that Mexican TTSuV1a sequences belong to genotype B, sharing phylogenetic origin, high nucleic acid and amino acid sequence similarity and dominant epitope conformation with commercially linked countries, such as the United States, Canada and China, whereas the Mexican TTSuV1b sequences belong to genotype A, being more divergent among each other and displaying low nucleotide identity with worldwide genotype A sequences. In both Iotatorquevirus species, a PTPase-like signature motif was identified in the predicted amino acid sequence, being more conserved for Mexican TTSuV1b sequences than for Mexican TTSuV1a sequences, in which several substitutions were observed. These changes may influence the conformation of dominant epitopes as different arrays were determined among TTSuV1a genotypes. ORF2 variability may account for pathogenic differences by modifying viral replication and immune response, as depicted for human TTV.

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Section: Discussionmentioning

confidence: 99%

Molecular characterization of the ORF2 of Torque teno sus virus 1a and Torque teno sus virus 1b detected in cases of postweaning multisystemic wasting syndrome in Mexico

Vargas-Ruiz¹,

García-Camacho²,

Álvarez³

et al. 2018

Transbound Emerg Dis

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“…A standard indirect Enzyme-linked immunosorbent assay (ELISA) using a purified rPotD protein-coated polystyrene microtiter 96-well plate (Costar; USA) and twofold serially diluted PotD antiserum in a range of 10 −2 -10 −5 was used to determine the neutralizing antibody IgG titers of the serum samples [Nieto et al, 2015]. The titer was defined as the absorbance of the highest dilution at a wavelength of 450 nm (A 450nm ) whose value was at least 0.1 above the average value of the background wells in which PBS replaced serum samples.…”

Section: Expression/purification Of Recombinant Potd Protein and Preparation Of Mouse Antiserummentioning

confidence: 99%

Deletion of Polyamine Transport Protein PotD Exacerbates Virulence in Glaesserella (Haemophilus) parasuis in the Form of Non-biofilm-generated Bacteria in a Murine Acute Infection Model

Dai

Yang

et al. 2021

Virulence

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Polyamines are small, polycationic molecules with a hydrocarbon backbone and multiple amino groups required for optimal cell growth. The potD gene, belonging to the ABC (ATP-binding cassette) transport system potABCD, encodes the bacterial substrate-binding subunit of the polyamine transport system, playing a pivotal role in bacterial metabolism and growth. The swine pathogen Glaesserella parasuis possesses an intact pot operon, and the studies presented here mainly examined the involvement of PotD in Glaesserella pathogenesis. A potD-deficient mutant was constructed using a virulent G. parasuis strain SC1401 by natural transformation; immuno-electron microscopy was used to identify the subcellular location of native PotD protein; an electron microscope was adopted to inspect biofilm and bacterial morphology; immunofluorescence technique was employed to study cellular adhesion, the levels of inflammation and apoptosis. The TSA++-pre-cultured mutant strain showed a significantly reduced adhesion capacity to PK-15 and MLE-12 cells. Likewise, we also found attenuation in virulence using murine models focusing on the clinical sign, H&E, and IFA for inflammation and apoptosis. However, when the mutant was grown in TSB++, virulence recovered to normal levels, along with a high level of radical oxygen species formation in the host. The expression of PotD could actively stimulate the production of ROS in Raw 264.7. Our data suggested that PotD from G. parasuis has a high binding potential to polyamine, and is essential for the full bacterial virulence within mouse models. However, the virulence of the potD mutant is highly dependent on its TSA++ culture conditions rather than on biofilm-formation.

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Anelloviruses

Segalés¹,

Kekarainen²

2019

Diseases of Swine

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Development of an indirect ELISA assay for the detection of IgG antibodies against the ORF1 of Torque teno sus viruses 1 and 2 in conventional pigs

Cited by 5 publications

References 38 publications

Molecular characterization of the ORF2 of Torque teno sus virus 1a and Torque teno sus virus 1b detected in cases of postweaning multisystemic wasting syndrome in Mexico

Molecular characterization of the ORF2 of Torque teno sus virus 1a and Torque teno sus virus 1b detected in cases of postweaning multisystemic wasting syndrome in Mexico

Deletion of Polyamine Transport Protein PotD Exacerbates Virulence in Glaesserella (Haemophilus) parasuis in the Form of Non-biofilm-generated Bacteria in a Murine Acute Infection Model

Anelloviruses

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