Evaluation of a multiplex real-time RT-PCR for quantitative and differential detection of wild-type viruses and C-strain vaccine of Classical swine fever virus

Jian-jun, Zhao; Cheng, Dan; Li, Na; Sun, Yuan; Shi, Zhu‐Pei; Zhu, Qing; Tu, Changchun; Tong, Guangzhi; Qiu, Hua‐Ji

doi:10.1016/j.vetmic.2007.04.046

Cited by 113 publications

(76 citation statements)

References 24 publications

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“…In particular, the mixed infection among CSFV, PRRSV, HP-PRRSV and PRV are often clinically manifested as porcine reproductive and respiratory syndrome, high fever etc. (Dong et al 2006, Zhao et al 2008, Zhou and Yang 2010, Zanella et al 2012. Therefore, it is difficult to accurately and timely distinguish these infections only based on clinical manifestations and pathological changes.…”

Section: Discussionmentioning

confidence: 99%

A multiplex PCR for simultaneous detection of classical swine fever virus, African swine fever virus, highly pathogenic porcine reproductive and respiratory syndrome virus, porcine reproductive and respiratory syndrome virus and pseudorabies in swines

Hu¹,

Lin²,

Yang³

et al. 2015

Polish Journal of Veterinary Sciences

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In this assay, we developed and evaluated a multiplex PCR (mPCR) for its ability in detecting multiple infections of swine simultaneously. Four pairs of primers were used to detect five viruses. Specific primers were designed for classical swine fever virus (CSFV), African swine fever virus (ASFV) and pseudorabies (PRV). A pair of primers was designed prudently for two different types of porcine reproductive and respiratory syndrome virus that respectively were porcine reproductive and respiratory syndrome virus (PRRSV), highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV). The detection limits of the mPCR were 1. A total of 49 clinical specimens were tested by the mPCR, and the result showed that co-infection by two or three viruses was 51%. In conclusion, the PCR is a useful tool for clinical diagnosis of not only single infections but also mixed infections in swines.

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Section: Discussionmentioning

confidence: 99%

A multiplex PCR for simultaneous detection of classical swine fever virus, African swine fever virus, highly pathogenic porcine reproductive and respiratory syndrome virus, porcine reproductive and respiratory syndrome virus and pseudorabies in swines

Hu¹,

Lin²,

Yang³

et al. 2015

Polish Journal of Veterinary Sciences

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“…The viral genome copies in CSFV-infected PK-15 cells were quantified by real-time reverse transcription-PCR (RT-PCR) (27). Briefly, the total RNA was extracted from CSFV-infected cells with TRIzol (catalog no.…”

Section: Methodsmentioning

confidence: 99%

“…The isolated RNA was then reverse transcribed to cDNA with Moloney murine leukemia virus reverse transcriptase (TaKaRa) according to the manufacturer's instructions. Quantification of genome copies of CSFV was performed as described previously (27). The expression of interleukin 8 (IL-8) was examined by a relative quantitative real-time RT-PCR.…”

Section: Methodsmentioning

confidence: 99%

Thioredoxin 2 Is a Novel E2-Interacting Protein That Inhibits the Replication of Classical Swine Fever Virus

Wang

et al. 2015

J Virol

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The E2 protein of classical swine fever virus (CSFV) is an envelope glycoprotein that is involved in virus attachment and entry. To date, the E2-interacting cellular proteins and their involvement in viral replication have been poorly documented. In this study, thioredoxin 2 (Trx2) was identified to be a novel E2-interacting partner using yeast two-hybrid screening from a porcine macrophage cDNA library. Trx2 is a mitochondrion-associated protein that participates in diverse cellular events. The Trx2-E2 interaction was further confirmed by glutathione S-transferase (GST) pulldown, in situ proximity ligation, and laser confocal assays. The thioredoxin domain of Trx2 and the asparagine at position 37 (N37) in the E2 protein were shown to be critical for the interaction. C lassical swine fever (CSF), caused by classical swine fever virus (CSFV), is a highly contagious, often fatal porcine disease with significant economic losses. CSFV belongs to the Pestivirus genus within the Flaviviridae family (1). It is an enveloped virus with a single-stranded, positive-sense RNA genome of approximately 12.3 kb in length. The RNA genome contains a single large open reading frame (ORF). This ORF is translated into a polyprotein, which is further processed into 12 mature proteins (N pro -C-E rns -E1-E2-p7-NS2-3-NS4A-NS4B-NS5A-NS5B) by viral and cellular proteases (2). The C, E rns , E1, and E2 proteins represent the structural components of the virion. The E1 and E2 glycoproteins are anchored to the envelope by their carboxyl termini, while E rns is loosely associated with the envelope. E rns presents as homodimers linked by disulfide bridges in virus-infected cells and in virions (3, 4). The C terminus of E2 functions as a membrane-spanning domain anchoring the E2-E1 or E2-E2 dimer to the viral envelope (5). E2 is also involved in virus attachment to and entry into the target cell (6). Furthermore, as a virulence determinant in pigs (7), the E2 protein can efficiently induce protective immune responses (8-13). Additionally, we recently showed that host ␤-actin interacts with E2 and modulates the early life cycle of CSFV (14).Thioredoxins (Trxs) are a class of ubiquitously expressed redox proteins that contain a conserved consensus amino acid sequence (Cys-Gly-Pro-Cys) in the catalytic center. There are two distinct forms of Trxs (Trx1 and Trx2), which act as antioxidants facilitating the reduction of other proteins by cysteine thiol-disulfide exchange. Trx2 consists of 157 amino acid (aa) residues, and its thioredoxin domain is located in the C terminus of the protein.Trxs are involved in diverse biological processes, such as cell growth, proliferation, apoptosis, and gene regulation (15-18). Additionally, they interact with several transcription factors and

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“…Different tissues, including brain, spleen, lung, kidney, urinary bladder, tonsils, and lymph nodes, were collected at 14 dpc and subjected to detection of the CSFV RNA by a real-time reverse transcription-PCR (RT-PCR) as described previously (21).…”

Section: Figmentioning

confidence: 99%

Generation and Efficacy Evaluation of a Recombinant Pseudorabies Virus Variant Expressing the E2 Protein of Classical Swine Fever Virus in Pigs

Wang

Yuan

et al. 2015

Clin Vaccine Immunol

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Classical swine fever (CSF) is an economically important infectious disease of pigs caused by classical swine fever virus (CSFV). Pseudorabies (PR), which is caused by pseudorabies virus (PRV), is another important infectious disease of pigs and other animals. Coinfections of pigs with PRV and CSFV occur occasionally in the field. The modified live vaccine Bartha-K61 strain has played an important role in the control of PR in many countries, including China. Since late 2011, however, increasing PR outbreaks caused by an emerging PRV variant have been reported in Bartha-K61-vaccinated swine populations on many farms in China. Previously, we generated a gE/gI-deleted PRV (rPRVTJ-delgE) based on this PRV variant, which was shown to be safe and can provide rapid and complete protection against lethal challenge with the PRV variant in pigs. Here, we generated a new recombinant PRV variant expressing the E2 gene of CSFV (rPRVTJ-delgE/gI-E2) and evaluated its immunogenicity and efficacy in pigs. The results showed that rPRVTJ-delgE/gI-E2 was safe for pigs, induced detectable anti-PRV and anti-CSFV neutralizing antibodies, and provided complete protection against the lethal challenge with either the PRV TJ strain or the CSFV Shimen strain. The data indicate that rPRVTJ-delgE/gI-E2 is a promising candidate bivalent vaccine against PRV and CSFV coinfections.

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Evaluation of a multiplex real-time RT-PCR for quantitative and differential detection of wild-type viruses and C-strain vaccine of Classical swine fever virus

Cited by 113 publications

References 24 publications

A multiplex PCR for simultaneous detection of classical swine fever virus, African swine fever virus, highly pathogenic porcine reproductive and respiratory syndrome virus, porcine reproductive and respiratory syndrome virus and pseudorabies in swines

A multiplex PCR for simultaneous detection of classical swine fever virus, African swine fever virus, highly pathogenic porcine reproductive and respiratory syndrome virus, porcine reproductive and respiratory syndrome virus and pseudorabies in swines

Thioredoxin 2 Is a Novel E2-Interacting Protein That Inhibits the Replication of Classical Swine Fever Virus

Generation and Efficacy Evaluation of a Recombinant Pseudorabies Virus Variant Expressing the E2 Protein of Classical Swine Fever Virus in Pigs

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