Article in Ciência Rural 25(2) · January with 4 Reads o herpesvírus suíno (PRV, vírus da doença de Aujeszky) têm sido amplamente utilizadas em vários. doença de Aujeszky em sistema de baculovirus. Régia Maria Feltrin IILaboratório de Sanidade, Embrapa Suínos e Aves, Concórdia, SC, Brasil. IIICentro de. CLONING AND EXPRESSION OF AUJESZKY’S DISEASE VIRUS GLYCOPROTEIN E .. vírus da doença de Aujeszky de surtos em suínos no Estado de Santa.
|Country:||Moldova, Republic of|
|Published (Last):||17 September 2013|
|PDF File Size:||11.51 Mb|
|ePub File Size:||9.44 Mb|
|Price:||Free* [*Free Regsitration Required]|
World Organization for Animal Health. This region was highly conserved for all reported genomes as shown by aligning of these sequences. Lanes 1 and 3 are amplification products, Lanes 2 and 4 are amplification products after digestion with Sma I.
The polymerase chain reaction PCR is a rapid tool that can be used no only to detect acutely PRV infected pigs but it is the recommended test aujesaky detect PRV latent infection. Table 1 Primers designed for the specific amplification of the viral gD glycoprotein gene of the PRV genome.
A rapid and accurate diagnosis of PRV infection is important for the initiation of appropriate control strategies. This assay was based on the amplification of a highly conserved viral gD gene fragment. In general, PRV infections must be considered in the differential diagnosis of respiratory, reproductive and nervous disorders.
Also, the BLAST search against nucleotide databases of different herpesvirus and random nucleotide sequences revealed this region is very specific for PRV genomes. The conventional diagnostic procedure is time-consuming and false-negative results may occur in submissions from latently infected animals.
Doença de Aujeszky
The effect of temperature and oligonucleotide primer length on the specificity and efficiency of amplification by the polymerase chain reaction.
The analysis directly from clinical samples from naturally infected animals proved the potential usefulness of the method for a rapid disease diagnosis from field cases. The etiological agent of this disease is suid herpesvirus type 1, usually named pseudorabies virus PRVa pantropic alphaherpesvirus which causes fatal infections in baby pigs, respiratory disease and poor growth in fattening pigs and reproductive disorders in adults 28 Each one of the nine tissue field samples from pigs diagnosed as PRV infected based on clinical signs and laboratory methods yielded the corresponding PRV amplified product when analyzed.
The trigeminal ganglion is the most consistent site for virus isolation, although latent virus is usually difficult to culture or even impossible 113 and PCR is the method recommended to detect viral genome present in this site. The viral agent following a primary replication can establish latent infection and develops a latency-reactivation infection which allows its perpetuation in pig populations 1012 However, this method is time-consuming and false negative results may occur in submissions from latently infected animals The nucleotide sequence amplified in this study corresponds to a bp fragment in the gD gene of the PRV genome The assay specificity was demonstrated by the absence of amplifications in all heterologous viruses evaluated and in tissue samples derived from seven healthy pigs.
The analysis of tissue homogenate samples from naturally infected animals proved the potential usefulness of the method for a rapid disease diagnosis from field cases.
A rapid, sensitive and specific PCR-based diagnostic assay to detect pseudorabies virus in clinical samples is provided. Diagnostic methods for detection of Classical swine fever virus—Status quo and new developments. Author information Article notes Copyright and License information Disclaimer.
Multiplex PCR for the simultaneous detection of pseudorabies virus, porcine cytomegalovirus, and porcine circovirus in pigs. Support Center Support Center. Agarose gel electrophoresis was used to detect PCR products.
Doença de Aujeszky – Wikipédia, a enciclopédia livre
Oligonucleotide primers and restriction endonuclease selection PRV specific primers were designed using the Oligo 6. Seven virus-negative tissues samples from clinically healthy animals were also included. Primers sequences, genome positions and the size of PCR products are shown in Table 1.
The annealing temperature and number of cycles were determined experimentally. Potential sites of virus latency associated with indigenous pseudorabies viruses in feral swine. Replication in the respiratory tract, central nervous system and reproductive organs is responsible for pathological changes causing different disorders The analytical sensitivity of the test was estimated to be 1. Iowa State University Press; Traditionally, PRV detection is based on direct virus isolation followed by confirmation using immunofluorescence, immunoperoxidase or neutralization tests with specific antiserum 2.
The virus primarily replicates in the respiratory tract, spreads along cranial nerves to the brains and via lymph and blood to internal organs, with the reproductive organs being affected.
In addition, positive amplifications were obtained in all the tissue samples, from PRV natural infected pigs, evaluated. Cell biological and molecular characteristics of pseudorabies virus infections in cell cultures and in pigs with emphasis on the respiratory tract.
Finally, nucleic acids from tissue homogenates samples derived from seven healthy pigs, and a non infected PK cell line were also tested showing no positive products data not shown. Under typical conditions of intensive swine production, several clinically similar viral diseases can occur which require laboratory differential diagnosis.
The Sma I restriction endonuclease site was used for additional specificity confirmation of the amplification products. Cell biological and molecular characteristics of pseudorabies virus infections in cell cultures and pigs with emphasis on respiratory tract.
Induction and inhibition of apoptosis by pseudorabies virus in the trigeminal ganglion during acute infection of swine. Can J Com Med.
The analytical sensitivity of the test was consistently observed to be 1. Thus, the optimal concentration of MgCl2 and primers were 1. This gene codes for an envelope glycoprotein named gD which plays and important role in binding cellular receptors and is critical for virus replication in different organs Detection of porcine circovirus type 2, porcine parvovirus and porcine pseudorabies virus from pigs with postweaning multisystemic wasting syndrome by multiplex PCR.
Moreover, the viral genomes of a related herpesvirus and other DNA genome porcine viruses as follow: Articles from Brazilian Journal of Microbiology are provided here courtesy of Elsevier.
The virus principally affects pigs, which are considered to be the natural host for PRV and the reservoir of the virus in nature, but also infects a broad range of wild and non-porcine mammals with the important exception of higher-order primates 8. PCR experiments were performed on serial ten-fold dilutions of a viral suspension of PRV isolate V with a titer of 10 6. Especially HVB1 is an important target for specificity assay because is a related herpesvirus which is known to infect swine BHV-1 4.
The PCR assay described here provides a rapid, highly sensitive, and cost-effective laboratory diagnosis for pseudorabies infections. Specificity of the PRV amplicons was furthermore confirmed by Sma I restriction endonuclease analysis which generated the two expected fragments of and bp in length Fig.
The polymerase chain reaction PCR can be used to identify PRV genomes in secretions or organ samples and although some PCR assays for PRV detection with different sensitivities have been reported 37915 there is no standard procedure recommended so far 2.