A host's immune system can be invaded by mycotoxin deoxynivalenol (DON) poisoning and porcine circovirus type 2 (PCV2) infections, which affect the host's natural immune function. Pro-inflammatory cytokines, IL-1β and IL-6, are important regulators in the process of natural immune response, which participate in inflammatory response and enhance immune-mediated tissue damage. Preliminary studies have shown that DON promotes PCV2 infection by activating the MAPK signaling pathway. Here, we explored whether the mRNA expression of IL-1β and IL-6, induced by the combination of DON and PCV2, would depend on the MAPK signaling pathway. Specific pharmacological antagonists U0126, SP600125 and SB203580, were used to inhibit the activities of ERK, JNK and p38 in the MAPK signaling pathway, respectively. Then, the mRNA expression of IL-1β and IL-6 in PK-15 cells was detected to explore the effect of the MAPK signaling pathway on IL-1β and IL-6 mRNA induced by DON and PCV2. The results showed that PK-15 cells treated with DON or PCV2 induced the mRNA expression of IL-1β and IL-6 in a time- and dose-dependent manner. The combination of DON and PCV2 has an additive effect on inducing the mRNA expression of IL-1β and IL-6. Additionally, both DON and PCV2 could induce the mRNA expression of IL-1β and IL-6 via the ERK and the p38 MAPK signal pathways, while PCV2 could induce it via the JNK signal pathway. Taken together, our results suggest that MAPKs play a contributory role in IL-1β and IL-6 mRNA expression when induced by both DON and PCV2.
- MeSH
- buněčné linie MeSH
- Circovirus * MeSH
- infekce viry čeledi Circoviridae genetika metabolismus MeSH
- interleukin-1beta genetika MeSH
- interleukin-6 genetika MeSH
- MAP kinasový signální systém účinky léků MeSH
- messenger RNA MeSH
- prasata MeSH
- trichotheceny toxicita MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Psittacine beak and feather disease (PBFD) is one of the most significant viral diseases in psittacine birds. The aim of the presented study was to develop a highly specific and sensitive TaqMan real-time PCR assay for universal detection of beak and feather disease virus (BFDV). Primers and a hydrolysis probe were selected on the highly conserved regions belonging to the ORF1 of the BFDV genome which were identified by aligning 814 genomic sequences downloaded from the GenBank database. The evaluation of the reaction parameters suggested a reaction efficiency of 97.1%, with consistent detection of 101 virus copies/μl of nucleic acid extract. The low values of standard deviation and coefficient of variation indicate a high degree of reproducibility and repeatability. The diagnostic applicability of the assay was proven on 36 BFDV positive and 107 negative specimens of psittacine origin representing 28 species. The assay showed a 100% ability to detect distinct genetic variants of the virus. Our data suggest that the presented TaqMan real-time PCR represents a specific, sensitive and reliable assay facilitating the molecular detection of BFDV.
- MeSH
- Circovirus genetika izolace a purifikace MeSH
- diagnostické techniky molekulární metody MeSH
- DNA primery genetika MeSH
- infekce viry čeledi Circoviridae diagnóza veterinární virologie MeSH
- kvantitativní polymerázová řetězová reakce metody MeSH
- nemoci ptáků diagnóza virologie MeSH
- oligonukleotidové sondy genetika MeSH
- ptáci MeSH
- reprodukovatelnost výsledků MeSH
- senzitivita a specificita MeSH
- veterinární lékařství metody MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
A loop-mediated isothermal amplification (LAMP) method with a real-time monitoring system was developed for the detection of porcine circovirus type 1 (PCV1) in commercial swine vaccines. This method was highly specific for PCV1. No cross-reaction to porcine circovirus type 2, porcine parvovirus, pseudorabies virus, classical swine fever virus, and porcine reproductive and respiratory syndrome virus was observed. The analytical sensitivity of the LAMP for PCV1 DNA was 10 copies/μl in the case of positive recombinant plasmid comparable to that obtained from the nested polymerase chain reaction (nested PCR). Furthermore, 25 commercial swine vaccines were tested by both the LAMP and the nested PCR, and three of them were tested positive for PCV1 DNA. These results indicate that PCV1 DNA can be real-time detected by the LAMP; the method was highly specific, sensitive, and rapid for the detection of PCV1 DNA, particularly in commercial swine vaccines.
- MeSH
- Circovirus klasifikace genetika izolace a purifikace MeSH
- DNA primery genetika MeSH
- infekce viry čeledi Circoviridae prevence a kontrola veterinární virologie MeSH
- nemoci prasat prevence a kontrola virologie MeSH
- prasata MeSH
- techniky amplifikace nukleových kyselin metody MeSH
- virové vakcíny genetika izolace a purifikace MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
A capsid protein of porcine circovirus 2 (PCV 2) serves as a diagnostic antigen for the detection of PCV 2-associated disease known as a postweaning multisystemic wasting syndrome (PMWS). In this report, a bacterial expression system was developed for the expression and purification of the full-length PCV 2 capsid (Cap) protein from a codon-optimized cap gene. Replacement of rare arginine codons located at the 5' end of the cap reading frame with codons optimal for E. coli was found to overcome the poor expression of the viral protein in the prokaryotic system. The Cap protein was purified to greater than 95% homogeneity by using a single cation-exchange chromatography at a yield of 10 mg per litre of bacterial culture. Despite the failure of the E. coli-expressed Cap protein to self-assemble into virus-like particles (VLPs), the immunization of mice with recombinant Cap yielded antibodies with the same specificity as those raised against native PCV 2 virions. In addition, the antigenic properties of the purified Cap protein were employed in a subunit-based indirect ELISA to monitor the levels of PCV 2 specific antibodies in piglets originating from a herd which was experiencing PCV 2 infection. These results pave the way for a straightforward large-scale production of the recombinant PCV 2 capsid protein and its use as a diagnostic antigen or a PCV 2 subunit vaccine.
- MeSH
- Circovirus genetika imunologie metabolismus MeSH
- Escherichia coli genetika metabolismus MeSH
- imunizace MeSH
- molekulární sekvence - údaje MeSH
- nemoci prasat diagnóza prevence a kontrola virologie MeSH
- prasata virologie MeSH
- protilátky virové krev MeSH
- rekombinantní proteiny genetika imunologie metabolismus MeSH
- sekvence aminokyselin MeSH
- syndrom multisystémového chřadnutí selat po odstavení diagnóza prevence a kontrola virologie MeSH
- virion metabolismus MeSH
- virové plášťové proteiny diagnostické užití genetika imunologie metabolismus MeSH
- virové vakcíny aplikace a dávkování imunologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- MeSH
- Circovirus patogenita MeSH
- DNA virů MeSH
- dospělí MeSH
- infekce viry čeledi Circoviridae diagnóza MeSH
- jaterní cirhóza chirurgie mikrobiologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- polymerázová řetězová reakce MeSH
- transplantace jater MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- přehledy MeSH
- srovnávací studie MeSH
- MeSH
- Circovirus genetika izolace a purifikace klasifikace MeSH
- DNA virů genetika MeSH
- infekce viry čeledi Circoviridae epidemiologie veterinární MeSH
- nemoci prasat epidemiologie virologie MeSH
- polymerázová řetězová reakce veterinární MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Geografické názvy
- Česká republika MeSH
- MeSH
- DNA viry patogenita MeSH
- genotyp MeSH
- hepatitida C epidemiologie terapie MeSH
- hepatitida virologie MeSH
- infekce viry čeledi Circoviridae etiologie přenos MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
