Detection of Asymptomatic Antigenemia in Pigs Infected by Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) by a Novel Capture Immunoassay with Monoclonal Antibodies against the Nucleocapsid Protein of PRRSV

Cai, Jian‐Piao; Wang, Yadi; Tse, Herman; Xiang, Hua; Yuen, Kwok‐Yung; Che, Xin

doi:10.1128/cvi.00244-09

Cited by 9 publications

(7 citation statements)

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“…Seventeen point eight percent (79/444) of the asymptomatic pigs were positive in this study. Similar findings (15.02%, 70/466) were reported in a study that employed traditional RT-PCR in China [ 29 ]. However, this is first study to report the viral load using serum samples in asymptomatic PRRSV-infected pigs using ZNA probe-based real-time PCR.…”

Section: Discussionsupporting

confidence: 88%

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Comparison of viremia of type II porcine reproductive and respiratory syndrome virus in naturally infected pigs by zip nucleic acid probe-based real-time PCR

Lin¹,

Lin²,

Hung³

et al. 2013

BMC Vet Res

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BackgroundPorcine reproductive and respiratory syndrome virus (PRRSV) is a RNA virus with high genetic variation. This virus causes significant economic losses in most pig-producing countries. The clinical presentation of PRRSV ranges from asymptomatic to devastating. In this study, we developed a sensitive and specific zip nucleic acid probe-based real-time PCR assay to evaluate the viremia of natural PRRSV-infected pigs in Taiwan. Serum samples were collected from 577 pigs aged 5–12 weeks. These include 444 clinically healthy pigs and 133 symptomatic pigs were confirmed to have porcine respiratory disease complex (PRDC).ResultsViremia was quantified in 79 of the 444 (17.8%) clinically healthy pigs and in 112 of the 133 (84.2%) PRDC cases. Viremias were significantly more common in pigs with PRDC compared with the clinically healthy pigs (P <0.0001). These results suggest that a high viral load is a major feature of PRRSV-affected pigs.ConclusionsZNA probe-based real-time PCR can be a useful tool to diagnose symptomatic and asymptomatic PRRSV-infected pigs. The presence of this marker in a sample of animals with high PRRSV loads (>104.2 PRRSV genomes/μl of serum) seems to indicate that it correlates with the presence of PRDC in pigs.

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Section: Discussionsupporting

confidence: 88%

“…The viral load is an indicator of active infection, virus-host interaction and disease progression [ 28 ]. The asymptomatic pigs are believed to serve as important reservoirs for the transmission of PRRSV to uninfected pigs [ 29 ]. Seventeen point eight percent (79/444) of the asymptomatic pigs were positive in this study.…”

Section: Discussionmentioning

confidence: 99%

Comparison of viremia of type II porcine reproductive and respiratory syndrome virus in naturally infected pigs by zip nucleic acid probe-based real-time PCR

Lin¹,

Lin²,

Hung³

et al. 2013

BMC Vet Res

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“…As illustrated in Figure 4B, the level of serum Abs specific to PRRSV was also determined at 0, 10 and 21 dpc by the IDEXX ELISA. This ELISA detects Abs mainly specific to the PRRSV N protein [54,55]. Pigs that had not received the inactivated vaccine developed N protein-specific Abs from 10 dpc with no differences in these Ab levels between groups.…”

Section: Resultsmentioning

confidence: 99%

Immunogenic and protective properties of GP5 and M structural proteins of porcine reproductive and respiratory syndrome virus expressed from replicating but nondisseminating adenovectors

Roques

Girard

St-Louis

et al. 2013

Vet Res

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Porcine reproductive and respiratory syndrome virus (PRRSV) is responsible for significant economic losses in the porcine industry. Currently available commercial vaccines do not allow optimal and safe protection. In this study, replicating but nondisseminating adenovectors (rAdV) were used for the first time in pigs for vaccinal purposes. They were expressing the PRRSV matrix M protein in fusion with either the envelope GP5 wild-type protein (M-GP5) which carries the major neutralizing antibody (NAb)-inducing epitope or a mutant form of GP5 (M-GP5m) developed to theoretically increase the NAb immune response. Three groups of fourteen piglets were immunized both intramuscularly and intranasally at 3-week intervals with rAdV expressing the green fluorescent protein (GFP, used as a negative control), M-GP5 or M-GP5m. Two additional groups of pigs were primed with M-GP5m-expressing rAdV followed by a boost with bacterially-expressed recombinant wild-type GP5 or were immunized twice with a PRRSV inactivated commercial vaccine. The results show that the rAdV expressing the fusion proteins of interest induced systemic and mucosal PRRSV GP5-specific antibody response as determined in an ELISA. Moreover the prime with M-GP5m-expressing rAdV and boost with recombinant GP5 showed the highest antibody response against GP5. Following PRRSV experimental challenge, pigs immunized twice with rAdV expressing either M-GP5 or M-GP5m developed partial protection as shown by a decrease in viremia overtime. The lowest viremia levels and/or percentages of macroscopic lung lesions were obtained in pigs immunized twice with either the rAdV expressing M-GP5m or the PRRSV inactivated commercial vaccine.

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“…It is known that NP is a highly conserved antigen among a variety of influenza viruses (18). Accordingly, isolation of conserved NPs from H9N2 viruses and preparation of monoclonal antibodies against NPs can be helpful in identifying the virus in different species, based on serological methods (19)(20)(21)(22) and competitive ELISA assay (23).…”

Section: Introductionmentioning

confidence: 99%

Monoclonal Antibody Preparation Against Nucleoprotein of Avian Influenza Virus Subtype H9N2

Hashemi

Madani

Aghamaali

et al. 2019

Iran Red Crescent Med J

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Background:In recent years, various poultry diseases have posed risks to this industry. Respiratory and infectious diseases are the most common diseases. According to previous findings, influenza disease is recognized as a significant life-threatening disease in the industry of poultry worldwide. Influenza virus type A, which belongs to the family Orthomyxoviridae, is responsible for a serious infectious disease. H9N2 AVI subtype circulates in the poultry worldwide, causing significant economic losses and infections in humans, also domestic and wild animals. Objectives: The purpose of the present experimental research was monoclonal antibodies (MAbs) production in contradiction of the NP of avian influenza virus (AIV) H9N2 subtype to detect AIV antigens and antibodies in the Department of Proteomics and Biochemistry, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran. in 2017. Methods: The conserved protein of NP from H9N2A/Chicken/Iran/259/2014 virus, with 60 kDa molecular weight, was isolated using the electroelution method. The purified protein was applied for Australian BALB/c mice immunization. After evaluating immunization by ELISA assay, the spleen of immunized mice was isolated and hybridized with SP2/0 myeloma cell. Next, the hybridized cell was cultured, and clone soups were collected after 15 days to examine antibodies via ELISA assay. The produced antibodies using Western blotting and antibody isotype kits were characterized.Results: Thirty antibody-producing clones were examined for reactivity against Nucleoprotein (NP), the antigen, at 1 µg/mL concentration. According to the ELISA assay of antibody titers, two (3/F10 and 2/D7) out of 30 antibodies were bound to the antigen with titer 0.863 and 1.641, respectively. Two hybrid clones, 2D7 and 3F10, which produced anti-NP antibodies, were isolated and cultured. Characterizing of the produced antibodies using Western blotting was performed using H9N2 virus; finally, two clone soups (3/F10 and 2/D7) reacted with the NP virus protein. According to the isotyping of antibodies produced by 3F10 and 2D7 clones, 3F10 clone produced IgG1 with a κ chain, and IgG1 concentration was 1.997, based on ELISA assay. Also, 2D7 clone produced IgG2a with a κ chain, and IgG2a concentration was obtained 1.951. Conclusions: According to the findings of our study, the produced antibody might be used in the diagnosis of influenza.

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Detection of Asymptomatic Antigenemia in Pigs Infected by Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) by a Novel Capture Immunoassay with Monoclonal Antibodies against the Nucleocapsid Protein of PRRSV

Abstract: These findings suggest that the antigen assay is a valuable field tool for the epidemiological control of PRRSV that can be used for rapid screening, particularly in asymptomatic animals.

Cited by 9 publications

References 35 publications

Comparison of viremia of type II porcine reproductive and respiratory syndrome virus in naturally infected pigs by zip nucleic acid probe-based real-time PCR

Comparison of viremia of type II porcine reproductive and respiratory syndrome virus in naturally infected pigs by zip nucleic acid probe-based real-time PCR

Immunogenic and protective properties of GP5 and M structural proteins of porcine reproductive and respiratory syndrome virus expressed from replicating but nondisseminating adenovectors

Monoclonal Antibody Preparation Against Nucleoprotein of Avian Influenza Virus Subtype H9N2

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