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1.
Virol J ; 21(1): 87, 2024 04 19.
Artigo em Inglês | MEDLINE | ID: mdl-38641833

RESUMO

BACKGROUND: Bovine parvovirus (BPV) is an autonomous DNA virus with a smaller molecular size and subtle differences in its structural proteins, unlike other animal parvoviruses. More importantly, this virus has the potential to produce visible to silent economic catastrophes in the livestock business, despite receiving very little attention. Parvoviral virus-like particles (VLPs) as vaccines and as logistical platforms for vaccine deployment are well studied. However, no single experimental report on the role of VP1 in the assembly and stability of BPV-VLPs is available. Furthermore, the self-assembly, integrity and stability of the VLPs of recombinant BPV VP2 in comparison to VP1 VP2 Cap proteins using any expression method has not been studied previously. In this study, we experimentally evaluated the self-assembling ability with which BPV virus-like particles (VLPs) could be synthesized from a single structural protein (VP2) and by integrating both VP2 and VP1 amino acid sequences. METHODS: In silico and experimental cloning methods were carried out. His-tagged and without-His-tag VP2 and V1VP2-encoding amino acid sequences were cloned and inserted into pFastbacdual, and insect cell-generated recombinant protein was evaluated by SDS‒PAGE and western blot. Period of infectivity and expression level were determined by IFA. The integrity and stability of the BPV VLPs were evaluated by transmission electron microscopy. The secondary structure of the BPV VLPs from both VP2 and V1VP2 was analyzed by circular dichroism. RESULTS: Our findings show that VP2 alone was equally expressed and purified into detectable proteins, and the stability at different temperatures and pH values was not appreciably different between the two kinds of VLPs. Furthermore, BPV-VP2 VLPs were praised for their greater purity and integrity than BPV-VP1VP2 VLPs, as indicated by SDS‒PAGE. Therefore, our research demonstrates that the function of VP1 has no bearing on the stability or integrity of BPV-VLPs. CONCLUSIONS: In summary, incredible physiochemically stable BPV VP2-derived VLPs have been found to be promising candidates for the development of multivalent vaccines and immunodiagnostic kits against enteric viruses and to carry heterogeneous epitopes for various economically important livestock diseases.


Assuntos
Bocavirus , Parvovirus , Vacinas , Animais , Baculoviridae/genética , Proteínas Recombinantes/genética , Proteínas do Capsídeo/genética
2.
Arch Microbiol ; 200(6): 841-846, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29846759

RESUMO

Virus infections are the root cause of epidemics in the world. Vaccines and antiviral agents have been the two important methods to control viral diseases; in recent times, RNA-mediated therapeutics and prevention have received much attention. In this review, we provide an overview of the current information regarding the use of vaccines, antiviral agents, and RNA-mediated methods in controlling or preventing viral infections. We stress specifically on the potential of existing RNA-mediated methods in clinical applications.


Assuntos
Antivirais/farmacologia , Descoberta de Drogas/tendências , Viroses/virologia , Vírus/efeitos dos fármacos , Animais , Humanos , RNA Viral/genética , Viroses/tratamento farmacológico , Fenômenos Fisiológicos Virais/efeitos dos fármacos , Vírus/genética
3.
Genomics ; 107(4): 150-4, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26826341

RESUMO

Long noncoding (lnc)RNAs comprise a diverse group of transcripts including large intervening noncoding (linc)RNAs, natural antisense transcripts (NATs) and intronic lncRNAs. The functions and mechanisms of more than 200 lncRNAs have been studied in vitro and the results suggest that lncRNAs may be molecular markers of prognosis in cancer patients. Some lncRNAs can promote virus replication and allow escape from cytosolic surveillance to suppress antiviral immunity. For example, lncRNA can cause persistent infection by Theiler's virus, and microRNA (miR)-27a/b is important for efficient murine cytomegalovirus (MCMV) replication. The available evidence suggests that lncRNAs may be potential targets of novel antiviral drugs.


Assuntos
RNA Longo não Codificante/genética , Replicação Viral , Vírus , Adenovírus Humanos/fisiologia , Animais , Humanos , Íntrons , Camundongos , Muromegalovirus/fisiologia , Theilovirus/fisiologia
4.
Virol J ; 10: 78, 2013 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-23497282

RESUMO

Foot-and-mouth disease (FMD) is one of most contagious animal diseases. It affects millions of cloven-hoofed animals and causes huge economic losses in many countries of the world. There are seven serotypes of which three (O, A and Asia 1) are endemic in China. Efficient control of FMD in China is crucial for the prevention and control of FMD in Asia and throughout the world. For the control of FMD, a powerful veterinary administration, a well-trained veterinary staff, a system of rapid and accurate diagnostic procedures and, in many countries, compulsory vaccination of susceptible animals are indispensable. This article strives to outline the Chinese animal disease control and prevention system, in particular for FMD, with the emphasis on diagnostic procedures applied in Chinese laboratories. In addition, new technologies for FMD diagnosis, which are currently in the phase of development or in the process of validation in Chinese laboratories, are described, such as lateral flow devices (LFD), Mab-based ELISAs, reverse transcription loop-mediated isothermal amplification (RT-LAMP) and gold nanopariticle immuno-PCR (GNP-IPCR).


Assuntos
Controle de Doenças Transmissíveis/métodos , Técnicas e Procedimentos Diagnósticos/veterinária , Febre Aftosa/diagnóstico , Febre Aftosa/prevenção & controle , Animais , China/epidemiologia , Controle de Doenças Transmissíveis/economia , Controle de Doenças Transmissíveis/instrumentação , Técnicas e Procedimentos Diagnósticos/economia , Técnicas e Procedimentos Diagnósticos/instrumentação , Febre Aftosa/epidemiologia , Febre Aftosa/virologia , Vírus da Febre Aftosa/genética , Vírus da Febre Aftosa/isolamento & purificação , Vírus da Febre Aftosa/fisiologia , Gado/virologia
5.
Virus Genes ; 46(2): 271-9, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23161403

RESUMO

The synonymous codon usage pattern of African swine fever virus (ASFV), the similarity degree of the synonymous codon usage between this virus and some organisms and the synonymous codon usage bias for the translation initiation region of viral functional genes in the whole genome of ASFV have been investigated by some simply statistical analyses. Although both GC12% (the GC content at the first and second codon positions) and GC3% (the GC content at the third codon position) of viral functional genes have a large fluctuation, the significant correlations between GC12 and GC3% and between GC3% and the first principal axis of principle component analysis on the relative synonymous codon usage of the viral functional genes imply that mutation pressure of ASFV plays an important role in the synonymous codon usage pattern. Turning to the synonymous codon usage of this virus, the codons with U/A end predominate in the synonymous codon family for the same amino acid and a weak codon usage bias in both leading and lagging strands suggests that strand compositional asymmetry does not take part in the formation of codon usage in ASFV. The interaction between the absolute codon usage bias and GC3% suggests that other selections take part in the formation of codon usage, except for the mutation pressure. It is noted that the similarity degree of codon usage between ASFV and soft tick is higher than that between the virus and the pig, suggesting that the soft tick plays a more important role than the pig in the codon usage pattern of ASFV. The translational initiation region of the viral functional genes generally have a strong tendency to select some synonymous codons with low GC content, suggesting that the synonymous codon usage bias caused by translation selection from the host takes part in modulating the translation initiation efficiency of ASFV functional genes.


Assuntos
Vírus da Febre Suína Africana/genética , Febre Suína Africana/virologia , Códon , Iniciação Traducional da Cadeia Peptídica , Proteínas Virais/genética , Vírus da Febre Suína Africana/classificação , Animais , Composição de Bases , Dados de Sequência Molecular , Suínos
6.
Virus Genes ; 44(3): 475-81, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22395914

RESUMO

Classical swine fever virus, bovine viral diarrhea virus (BVDV), and border disease virus can cause serious livestock diseases. The relative synonymous codon usage value, the "effective number of codons" (ENC), the ratio of K(s) value to K(a) value and the principle component analysis were employed to analyze the genetic characteristics of open reading frame (ORF) and the four genes (the N(pro), Erns, E1, E2 genes) of the three viruses and the relationship of codon usage pattern between each virus and its most common host. The amount of under-represented codons is larger than the amount of over-represented ones in ORFs or the four genes of the three viruses. The ENC value and the ratio of K(s)/K(a) for each gene show that mutation pressure plays a role in their evolutional processes. In addition, the evidence that selection from the natural host might influences the codon usage patterns of virus is found in the differences of codon usage patterns of ORF and Erns gene of BVDV strain ZM-95 isolated from domestic pig and those of the rest of BVDV strains isolated from cattle. These results indicate that although a strong mutation pressure from the three pestiviruses takes part in their evolutional processes by the alternation of synonymous codons, translation selection from the susceptible livestock on some genes should not be ignored. The codon usage pattern of the three pestiviruses is a result caused by the equilibrium of mutation pressure from virus and translation selection from its host.


Assuntos
Vírus da Doença da Fronteira/genética , Vírus da Febre Suína Clássica/genética , Códon , Vírus da Diarreia Viral Bovina Tipo 1/genética , RNA Viral/genética , Proteínas Virais/genética , Animais , Vírus da Doença da Fronteira/isolamento & purificação , Bovinos , Vírus da Febre Suína Clássica/isolamento & purificação , Biologia Computacional , Vírus da Diarreia Viral Bovina Tipo 1/isolamento & purificação , Gado , Fases de Leitura Aberta , Análise de Sequência de DNA , Sus scrofa
7.
Virol J ; 8: 419, 2011 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-21888667

RESUMO

BACKGROUND: FMD is one of the major causes of economic loss of cloven-hoofed animals in the world today. The assessment of dominant genotype/lineage and prevalent trends and confirmation the presence of infection or vaccination not only provides scientific basis and first-hand information for appropriate control measure but also for disease eradication and regaining FMD free status following an outbreak. Although different biological and serological approaches are still applied to study this disease, ELISA test based on the distinct format, antigen type and specific antibody reinforce its predominance in different research areas of FMD, and this may replace the traditional methods in the near future. This review gives comprehensive insight on ELISA currently available for typing, antigenic analysis, vaccination status differentiation and surveillance vaccine purity and content at all stages of manufacture in FMDV. Besides, some viewpoint about the recent advances and trends of ELISA reagent for FMD are described here. METHODS: More than 100 studies regarding ELISA method available for FMD diagnosis, antigenic analysis and monitor were thoroughly reviewed. We investigated previous sagacious results of these tests on their sensitivity, specificity. RESULTS: We found that in all ELISA formats for FMD, antibody-trapping and competitive ELISAs have high specificity and RT-PCR (oligoprobing) ELISA has extra sensitivity. A panel of monoclonal antibodies to different sites or monoclonal antibody in combination of antiserum is the most suitable combination of antibodies in ELISA for FMD. Even though from its beginning, 3ABC is proven to be best performance in many studies, no single NSP can differentiate infected from vaccinated animals with complete confidence. Meanwhile, recombinant antigens and peptide derived from FMDV NPs, and NSPs have been developed for use as an alternative to the inactivated virus antigen for security. CONCLUSIONS: There is a need of target protein, which accurately determines the susceptible animal status based on the simple, fast and reliable routine laboratory test. A further alternative based on virus-like particle (VLP, also called empty capsids) in combination of high throughput antibody technique (Phage antibody library/antibody microarray) may be the powerful ELISA diagnostic reagents in future.


Assuntos
Anticorpos Antivirais/imunologia , Antígenos Virais/análise , Doenças dos Bovinos/diagnóstico , Impressões Digitais de DNA/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Vírus da Febre Aftosa/genética , Febre Aftosa/diagnóstico , Doenças dos Suínos/diagnóstico , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/metabolismo , Anticorpos Antivirais/metabolismo , Antígenos Virais/imunologia , Bovinos , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/virologia , Diagnóstico Diferencial , Erradicação de Doenças , Ensaio de Imunoadsorção Enzimática/normas , Ensaio de Imunoadsorção Enzimática/tendências , Febre Aftosa/imunologia , Febre Aftosa/prevenção & controle , Febre Aftosa/virologia , Vírus da Febre Aftosa/classificação , Vírus da Febre Aftosa/imunologia , Camundongos , Sensibilidade e Especificidade , Suínos , Doenças dos Suínos/imunologia , Doenças dos Suínos/virologia , Estomatite Vesicular/imunologia , Vacinas Virais
8.
Virol J ; 8: 148, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21453461

RESUMO

BACKGROUND: Foot-and-mouth disease (FMD) is one of the most contagious of all artiodactyl animal diseases, and its infection has an obvious ability to spread over long distances and to contribute to epidemics in FMD-free areas. A highly sensitive and specific method is required to detect FMDV. In this study, we evaluated the usefulness of a bio-barcode assay (BCA) technique for detecting clinical samples of FMDV. METHODS: Highly sensitive gold nanopariticle (GNP) improved immuno -PCR (GNP-IPCR) which derived from the bio-barcode assay (BCA) was designed for the detection of FMDV. The target viral particles were captured by a polyclonal antibody coated on ELISA microplate, followed by adding GNP which was dually modified with oligonucleotides and a FMDV specific monoclonal antibody (MAb) 1D11 to form a sandwiched immune complex. After the formation of immuno-complex, the signal DNA was released by heating, and consequently characterized by PCR and real time PCR. RESULTS: The detection limit of GNP-PCR could reach to 10 fg/ml purified FMDV particles, and the assay can detect clinical samples of FMDV with highly sensitivity, while detect limit of conventional ELISA is 100 ng/ml in this study. CONCLUSION: GNP-IPCR may provide a highly sensitive method for the detection of FMDV.


Assuntos
Vírus da Febre Aftosa/isolamento & purificação , Febre Aftosa/diagnóstico , Ouro , Técnicas de Diagnóstico Molecular/métodos , Nanopartículas , Reação em Cadeia da Polimerase/métodos , Virologia/métodos , Animais , Febre Aftosa/virologia , Vírus da Febre Aftosa/genética , Vírus da Febre Aftosa/imunologia , Imunoensaio/métodos , Sensibilidade e Especificidade , Medicina Veterinária/métodos
9.
Virol J ; 8: 325, 2011 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-21708006

RESUMO

BACKGROUND: Cardioviruses are positive-strand RNA viruses in the Picornaviridae family that can cause enteric infection in rodents and also been detected at lower frequencies in other mammals such as pigs and human beings. The Cardiovirus genus consists two distinct species: Encephalomyocarditis virus (EMCV) and Theilovirus (ThV). There are a lot differences between the two species. In this study, the differences of codon usage in EMCV and ThV were compared. RESULTS: The mean ENC values of EMCV and ThV are 54.86 and 51.08 respectively, higher than 40.And there are correlations between (C+G)12% and (C+G)3% for both EMCV and ThV (r = -0.736; r = 0.986, P < 0.01, repectively). For ThV the (C+G)12%, (C+G)3%, axis f'1 and axis f'2 had a significant correlations respectively but not for EMCV. According to the RSCU values, the EMCV species seemed to prefer U, G and C ending codon, while the ThV spice seemed to like using U and A ending codon. However, in both genus AGA for Arg, AUU for Ile, UCU for Ser, and GGA for Gly were chosen preferentially. Correspondence analysis detected one major trend in the first axis (f'1) which accounted for 22.89% of the total variation, and another major trend in the second axis (f'2) which accounted for 17.64% of the total variation. And the plots of the same serotype seemed at the same region at the coordinate. CONCLUSION: The overall extents of codon usage bias in both EMCV and ThV are low. The mutational pressure is the main factor that determines the codon usage bias, but the (C+G) content plays a more important role in codon usage bias for ThV than for EMCV. The synonymous codon usage pattern in both EMCV and ThV genes is gene function and geography specific, but not host specific. Maybe the serotype is one factor effected the codon bias for ThV, and location has no significant effect on the variations of synonymous codon usage in these virus genes.


Assuntos
Aminoácidos/genética , Códon , Vírus da Encefalomiocardite/genética , Theilovirus/genética , Composição de Bases , Biologia Computacional/métodos , Mutação , Seleção Genética
10.
Virol J ; 8: 268, 2011 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-21635788

RESUMO

Foot-and-Mouth Disease (FMD), as a major global animal disease, affects millions of animals worldwide and remains the main sanitary barrier to the international and national trade of animals and animal products. Inactivated vaccination is the most effective measure for prevention of FMD at present, but fail to induce long-term protection and content new requires for production of FMD vaccines. As a number of Researchers hope to obtain satisfactory novel vaccines by new bio-technology, novel vaccines have been studied for more than thirty years. Here reviews the latest research progress of new vaccines, summarizes some importance and raises several suggestions for the future of FMD vaccine.


Assuntos
Febre Aftosa/prevenção & controle , Vacinas Virais/imunologia , Animais , Pesquisa Biomédica/tendências , Febre Aftosa/imunologia , Memória Imunológica , Vacinas de Produtos Inativados/imunologia
11.
Virol J ; 8: 146, 2011 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-21450075

RESUMO

BACKGROUND: Poliovirus, the causative agent of poliomyelitis, is a human enterovirus and a member of the family of Picornaviridae and among the most rapidly evolving viruses known. Analysis of codon usage can reveal much about the molecular evolution of the viruses. However, little information about synonymous codon usage pattern of polioviruses genome has been acquired to date. METHODS: The relative synonymous codon usage (RSCU) values, effective number of codon (ENC) values, nucleotide contents and dinucleotides were investigated and a comparative analysis of codon usage pattern for open reading frames (ORFs) among 48 polioviruses isolates including 31 of genotype 1, 13 of genotype 2 and 4 of genotype 3. RESULTS: The result shows that the overall extent of codon usage bias in poliovirus samples is low (mean ENC = 53.754 > 40). The general correlation between base composition and codon usage bias suggests that mutational pressure rather than natural selection is the main factor that determines the codon usage bias in those polioviruses. Depending on the RSCU data, it was found that there was a significant variation in bias of codon usage among three genotypes. Geographic factor also has some effect on the codon usage pattern (exists in the genotype-1 of polioviruses). No significant effect in gene length or vaccine derived polioviruses (DVPVs), wild viruses and live attenuated virus was observed on the variations of synonymous codon usage in the virus genes. The relative abundance of dinucleotide (CpG) in the ORFs of polioviruses are far below expected values especially in DVPVs and attenuated virus of polioviruses genotype 1. CONCLUSION: The information from this study may not only have theoretical value in understanding poliovirus evolution, especially for DVPVs genotype 1, but also have potential value for the development of poliovirus vaccines.


Assuntos
Composição de Bases , Códon , Poliovirus/genética , Sequência de Bases , Evolução Molecular , Humanos , Poliomielite/virologia , Poliovirus/classificação , Poliovirus/isolamento & purificação
12.
Arch Virol ; 156(1): 153-60, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21069395

RESUMO

Bovine viral diarrhea virus (BVDV) is a widespread virus in beef and dairy herds. BVDV has been grouped into two genotypes, genotype 1 and genotype 2. In this study, the relative synonymous codon usage (RSCU) values, effective number of codon (ENC) values and nucleotide content were investigated, and a comparative analysis of codon usage patterns for open reading frames (ORFs) of 22 BVDV genomes, including 14 of genotype 1 and 8 of genotype 2, was carried out. A high A+U content and low codon bias were found in BVDV genomes. Depending on the RSCU data, it was found that there was a significant variation in bias of codon usage between the two genotypes, and a geographic factor exists only in genotype-1 of BVDV. The RSCU data have a negative correlation with general average hydrophobicity (GRAVY), aromaticity and nucleotide content. Furthermore, the overall abundance of C and U has no effect on the synonymous codon usage patterns. In contrast, the A and G content showed a significant correlation with the nucleotide content at the third position. In addition, the codon usage patterns of BVDV are similar to those of 22 conserved genes of Bos taurus. Taken together, the genetic characteristics of BVDV possibly result from interactions between natural selection and mutation pressure.


Assuntos
Códon/genética , Vírus da Diarreia Viral Bovina/genética , Regulação Viral da Expressão Gênica/fisiologia , Animais , Genoma Viral , Fases de Leitura Aberta
13.
Virus Genes ; 42(2): 245-53, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21249440

RESUMO

In this study, the relative synonymous codon usage (RSCU) values, effective number of codon (ENC) values, nucleotide contents, and dinucleotide were used to investigate codon usage pattern of each protein-coding gene and genome among 31 Newcastle disease virus (NDV) isolates. The result shows that the overall extent of codon usage bias in NDV is low (mean ENC = 56.15 > 40). The good correlation between the (C + G)(12)% and (G + C)(3)% suggests that the mutational pressure, rather than natural selection, is the main factor that determines the codon usage bias and base component in NDV. It is observed that synonymous codon usage pattern in NDV genes is gene function and geography specific, but not host specific. By contrasting synonymous codon usage patterns of different NDV isolates, we suggest that more than one genotype of NDV circulates in waterfowl in USA; and gene length has no significant effect on the variations of synonymous codon usage in these virus genes. CpG under-represented is a characteristic for NDV to fit in its host. These results not only provide an insight into the variation of codon usage pattern among the genomes of NDV, but also may help in understanding the processes governing the evolution of NDV.


Assuntos
Códon/genética , Vírus da Doença de Newcastle/genética , Composição de Bases , Códon/análise , Evolução Molecular , Genoma Viral , Modelos Lineares , Mutação , Análise de Componente Principal , RNA Viral/genética
14.
Mol Cell Probes ; 24(2): 104-6, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19835950

RESUMO

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay targeting the nucleocapsid phosphoprotein gene of infectious bronchitis virus (IBV) was developed. The detection limits for the IBV RT-LAMP assay were 10(1) 50% egg infection dose (EID(50)) per 50 microl of titrated viruses and no cross-reaction of IBV RT-LAMP was found when tested with other viruses including Newcastle disease virus (NDV), avian reovirus (ARV), and infectious laryngotrachietis virus (ILTV) due to their mismatch with IBV RT-LAMP primers. A total of 187 clinical tissues samples (88 blood, 62 kidney and 37 lung) were evaluated and compared to conventional RT-PCR. The sensitivity of RT-LAMP and RT-PCR assays for detecting IBV RNA in clinical specimens was 99.5% and 98.4%, respectively. These findings showed that the RT-LAMP assay has potential usefulness for rapid and sensitive diagnosis in outbreak of IBV.


Assuntos
Galinhas/virologia , Infecções por Coronavirus/veterinária , Vírus da Bronquite Infecciosa/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Doenças das Aves Domésticas/virologia , Transcrição Reversa/genética , Temperatura , Animais , Sequência de Bases , Infecções por Coronavirus/virologia , Primers do DNA , Eletroforese em Gel de Ágar , Vírus da Bronquite Infecciosa/genética , Dados de Sequência Molecular , Especificidade de Órgãos , Sensibilidade e Especificidade
15.
Mol Cell Probes ; 23(2): 71-4, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19103283

RESUMO

The usefulness of reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid pre-clinical detection of classical swine fever virus (CSFV) infection was evaluated. The RT-LAMP reaction could be finished in 60 min under isothermal condition at 65 degrees C by employing a set of four primers targeting the 5' untranslated region of CSFV. The RT-LAMP assay of CSFV showed higher sensitivities than that of RT-PCR, with a detection limit of 5 copies per reaction. No cross-reactivity was observed from the samples of other related viruses including porcine circovirus type 2, porcine parvovirus, porcine pseudorabies virus, Japanese encephalitis virus, and porcine reproductive and respiratory syndrome virus. The detection rates of CSFV RT-LAMP, RT-PCR and virus isolation for samples including blood, tonsil, nasal and rectal swabs from uninoculated pigs without any clear clinical symptom were 89%, 78% and 71%, respectively. Furthermore, all of the assays showed higher sensitivity for blood and tonsil swabs samples than nasal and rectal swabs. These results indicate that the CSFV RT-LAMP assay is a valuable tool for its rapid, cost-effective detection and has potential usefulness for rapid pre-clinical detection and surveillance of classical swine fever in developing countries.


Assuntos
Vírus da Febre Suína Clássica/genética , Vírus da Febre Suína Clássica/isolamento & purificação , Peste Suína Clássica/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/métodos , Animais , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Sensibilidade e Especificidade , Suínos
16.
Vet Res Commun ; 32(6): 491-8, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18481190

RESUMO

Avian influenza and Newcastle disease are the highly contagious and most economically important diseases in poultry industry throughout the world. A multiplex reverse transcription polymerase chain reaction (mRT-PCR) assay was developed for the rapid and specific discrimination of H5 and H9 subtypes of avian influenza viruses (AIV) and Newcastle disease virus (NDV). Three sets of specific primers were applied in the assay based on the sequences of the hemagglutinin gene of H5-AIV, H9-AIV and fusion protein gene of NDV. 59 clinical samples including the throat washes, oral swabs, and cloacal scrapings were detected by mRT-PCR and single RT-PCR (sRT-PCR), respectively. The results indicated that the sensitivity and specificity of mRT-PCR were in accordance with sRT-PCR. The mRT-PCR developed in this study may therefore provide a new avenue to rapid detection of these important pathogens in one reaction.


Assuntos
Virus da Influenza A Subtipo H5N1/classificação , Vírus da Influenza A Subtipo H9N2/classificação , Influenza Aviária/virologia , Doença de Newcastle/virologia , Vírus da Doença de Newcastle/classificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Animais , Galinhas/virologia , Patos/virologia , Virus da Influenza A Subtipo H5N1/genética , Vírus da Influenza A Subtipo H9N2/genética , Vírus da Doença de Newcastle/genética , Sensibilidade e Especificidade , Organismos Livres de Patógenos Específicos
17.
J Vet Res ; 62(4): 431-437, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30729199

RESUMO

INTRODUCTION: The extremely high genetic variation and the continuously emerging variants of foot-and-mouth disease virus (FMDV) of Southern African Territory (SAT) serotypes including SAT1, SAT2, and SAT3 make it necessary to develop a new RT-PCR for general use for monitoring viruses based on the updated genome information. MATERIAL AND METHODS: A FMDV SAT-D8 one-step RT-PCR was established based on the 1D2A2B genes of the SAT serotype viruses with a multiplex primer set. FMDV A, O, C, and Asia 1 serotypes, other vesicular disease viruses, inactivated SAT viruses, and 125 bovine, ovine, caprine and porcine tissue samples collected from the Chinese mainland were included for evaluating the assay. RESULTS: The new RT-PCR was proven to be specific without cross-reactions with Eurasian FMDV, swine vesicular disease virus (SVDV), Seneca valley virus (SVV), or other common viral pathogens of cattle, sheep, goat, and pig. An around 257 bp-sized amplicon clearly appeared when the inactivated SAT viruses were detected. However, all 125 samples collected from FMDV-susceptible animals from the Chinese mainland which has not known SAT epidemics showed negative results. CONCLUSIONS: A FMDV SAT-D8 one-step RT-PCR is a promising method for primary screening for FMDV SAT serotypes.

18.
J Thorac Dis ; 8(6): 1069-79, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27293822

RESUMO

BACKGROUND: The cellular and molecular mechanisms underlying lung allograft rejection remain poorly understood. We investigated the potential role of interleukin (IL)-17A in lung transplant rejection in a mouse model, because previous studies in clinical and rodent models have implicated IL-17A in both acute and chronic rejection. METHODS: To generate an orthotopic lung transplantation model, lungs from C57BL/6 or BALB/c mice were transplanted into C57BL/6 mice (isograft and allograft models, respectively). The effects of anti-IL-17A treatment in allograft recipients were investigated. The histological features and rejection status of isografts and allografts were assessed at 3, 7, and 28 days after transplantation, and differences in graft infiltrating cells and mRNA expression of relevant cytokines were quantified at 3 and 7 days after transplantation. RESULTS: As expected, isografts showed no obvious signs of rejection, whereas allografts exhibited minimal-to-mild rejection (grade A1-A2) by day 3 and moderate-to-severe rejection (grade A3-A4) by day 7, without evidence of obliterative bronchiolitis (OB). However, by 28 days, evidence of OB was observed in 67% (2/3) of allografts and severe rejection (grade A4) was observed in all. IL-17 mRNA expression in allografts was increased with rejection, and interferon (IFN)-γ and IL-6 mRNA expression levels followed a similar pattern. In contrast, IL-22 expression in allografts was only slightly increased. Antibody (Ab) neutralization of IL-17A diminished the signs of acute rejection at 7 days after transplantation in allografts, and this early protection was accompanied by a decrease in cellular stress according to histological evaluation, suggesting the involvement of IL-17A in the development of early post-transplantation lesions. CONCLUSIONS: Our data indicate that IL-17A is important in the pathophysiology of allograft rejection, and neutralization of IL-17A is a potential therapeutic strategy to preventing lung transplant rejection.

19.
Infect Genet Evol ; 28: 101-6, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25152485

RESUMO

The 2A region of the foot-and-mouth disease virus (FMDV) polyprotein is 18 amino acids in length, and 2A self-cleavage site (2A/2B) contains a conserved amino acid motif G2A/P2B. To investigate the synonymous codon usage for Glycine at the 2A/2B cleavage site of FMDV, 66 2A/2B1 nucleotide sequences were aligned and found that the synonymous codon usage of G2A is conserved and GGG was the most frequently used. To examine the role of synonymous codons for G2A in self-cleavage efficiency of 2A/2B, recombinant constructs which contains the chloramphenicol acetyltransferase protein (CAT) and enhanced green fluorescent protein (EGFP) linked by the FMDV 2A sequence with four synonymous codons for G2A were produced. The activities of all the F2As based plasmids were determined in CHO cells. The results showed that the synonymous codon usage patterns for G2A at the cleavage site (2A/2B) have no effect on the cleavage efficiency. This suggests that the synonymous codon usage of 2A peptide has no effect on the cleavage efficiency of FMDV 2A element.


Assuntos
Vírus da Febre Aftosa/genética , Mutação Puntual , Proteínas Virais/genética , Motivos de Aminoácidos , Animais , Sequência de Bases , Linhagem Celular , Vírus da Febre Aftosa/classificação , Genótipo , Matrizes de Pontuação de Posição Específica , Sorogrupo , Proteínas Virais/química
20.
J Vet Sci ; 15(3): 423-6, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24690607

RESUMO

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed to rapidly detect foot-and-mouth disease virus serotype C (FMDV C). By testing 10-fold serial dilutions of FMDV C samples, sensitivity of the FMDV C RT-LAMP was found to be 10 times higher than that of conventional reverse transcription- PCR (RT-PCR). No cross-reactivity with A, Asia 1, or O FMDV or swine vesicular disease virus (SVDV) indicated that FMDV C RT-LAMP may be an exciting novel method for detecting FMDV C.


Assuntos
Febre Aftosa/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/métodos , Animais , Vírus da Febre Aftosa/genética , Técnicas de Amplificação de Ácido Nucleico/veterinária , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Transcrição Reversa/genética , Sensibilidade e Especificidade
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