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Maria Gabriela Echeverria - One of the best experts on this subject based on the ideXlab platform.
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expression and purification of Suid Herpesvirus 1 glycoprotein e in the baculovirus system and its use to diagnose aujeszky s disease in infected pigs
Protein Expression and Purification, 2013Co-Authors: Maria Soledad Serena, German Ernesto Metz, Eduardo Carlos Mortola, Christoph Geisler, Santiago Corva, Alejandra Larsen, Donald L Jarvis, Maria Gabriela EcheverriaAbstract:Suid Herpesvirus 1 (SHV-1) is the etiological agent of Aujeszky’s disease (AD), which affects swine herds worldwide and causes substantial economic losses due to animal mortality and lost productivity. In order to eradicate SHV-1, vaccination programs using viruses lacking the gene encoding glycoprotein E (gE) are ongoing in several countries. These eradication programs have generated a currently unmet demand for affordable and sensitive tests that can detect SHV-1 infection, yet distinguish between infected and vaccinated pigs. To meet this demand, we used the baculovirus-insect cell system to produce immunologically authentic full-length recombinant gE protein for use in a serum ELISA assay. As previous efforts to clone the gE gene had failed due to its extremely high GC-content (75% average), we used betaine as a PCR enhancer to facilitate amplification of the entire gE gene from the Argentinian CL15 strain of SHV-1. The cloned gE gene was expressed at high levels in recombinant baculovirus-infected insect cells and reacted strongly with sera from SHV-1 infected pigs. We used the recombinant gE protein to develop a local indirect ELISA test with sensitivity and specificity comparable to currently available commercial tests. Thus, recombinant gE produced in baculovirus-infected insect cells is a viable source of antigen for the detection of SHV-1 in ELISA tests. We also provide evidence supporting a potential application of this recombinant form of gE as a SHV-1 subunit vaccine.
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phylogenetic analysis of Suid Herpesvirus 1 isolates from argentina
Veterinary Microbiology, 2011Co-Authors: Maria Soledad Serena, German Ernesto Metz, Eduardo Carlos Mortola, Maria Gabriela EcheverriaAbstract:Argentinean Suid Herpesvirus 1 isolates were compared with reference strains and sequences available at GenBank and phylogenetically analyzed. A short fragment of the gE gene of the immunodominant epitopes was used for preliminary grouping of isolates by phylogenetic analysis. The analysis of the partial gC gene provided more precise genetic typing and segregation into the main genotypes I and II. Results confirmed that the Argentinean genotype I isolates predominate in our country. The topology of the partial gC gene was similar to that previously reported. The Argentinean type I isolates belonged to one cluster and grouped together with NIA-3 and American and Brazilian genotype I strains. However, the results obtained by the algorithms allow inferring that the Yamagata S-81 and Mer (genotype II) strains are grouped together.
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characterization of Suid Herpesvirus 1 field isolates from argentina
Revista Argentina De Microbiologia, 2010Co-Authors: Maria Soledad Serena, German Ernesto Metz, Eduardo Carlos Mortola, G Martin P Ocampos, S E Gambaro, Maria Gabriela EcheverriaAbstract:The genomic characterization of Suid Herpesvirus 1 (SHV-1) isolates from Argentina was accomplished by restriction pattern analysis using the BamHI, BstEII and XhoI enzymes. Type II genome has been described only once in Argen- tina. This study revealed considerable homogeneity of BamHI endonuclease sites in all the strains analyzed, accord- ing to the number and size of the fragments. No deletion of BamHI fragment #7 among the Argentinean isolates suggests that these strains are wild-type. In addition, the main antigenic domain of glycoprotein E of all the Argentinean strains, as well as the reference strains and sequences available in the GenBank, were characterized. The similarity percent oscillated between 99 and 100%.
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Characterization of Suid Herpesvirus 1 field isolates from Argentina Caracterización de cepas de campo de Herpesvirus suino 1 aisladas en Argentina
Elsevier España S.L.U., 2010Co-Authors: Maria Soledad Serena, German Ernesto Metz, Eduardo Carlos Mortola, G Martin P Ocampos, S E Gambaro, Maria Gabriela EcheverriaAbstract:The genomic characterization of Suid Herpesvirus 1 (SHV-1) isolates from Argentina was accomplished by restriction pattern analysis using the BamHI, BstEII and XhoI enzymes. Type II genome has been described only once in Argentina. This study revealed considerable homogeneity of BamHI endonuclease sites in all the strains analyzed, according to the number and size of the fragments. No deletion of BamHI fragment #7 among the Argentinean isolates suggests that these strains are wild-type. In addition, the main antigenic domain of glycoprotein E of all the Argentinean strains, as well as the reference strains and sequences available in the GenBank, were characterized. The similarity percent oscillated between 99 and 100%.Se caracterizaron cepas argentinas de Herpesvirus suino tipo 1 (HVS-1) mediante corte con las enzimas de restricción BamHI, BstEII y XhoI. La presencia del tipo genómico II había sido reportada en la Argentina una sola vez y, hasta el momento, no se han informado nuevos casos. Este estudio reveló una homogeneidad de los sitios BamHI, acorde con el número y el tamaño de los fragmentos. La presencia del fragmento BamHI #7 en los aislamientos argentinos analizados sugiere que éstos pertenecen al tipo salvaje (wild-type). Además, se caracterizó el principal dominio inmunogénico de la glicoproteína E de todas las cepas argentinas y se lo comparó con los de las cepas de referencia y con las secuencias disponibles en la base de datos GenBank. Los porcentajes de similitud obtenidos oscilaron entre 99 y 100%
Echeverría, María Gabriela - One of the best experts on this subject based on the ideXlab platform.
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Caracterización de cepas de campo de Herpesvirus suino 1 aisladas en Argentina
2020Co-Authors: Serena, María Soledad, Metz, Germán Ernesto, Martín Ocampos, Giselle Paula, Gambaro, Sabrina Eliana, Mórtola, Eduardo Carlos, Echeverría, María GabrielaAbstract:Se caracterizaron cepas argentinas de Herpesvirus suino tipo 1 (HVS-1) mediante corte con las enzimas de restricción BamHI, BstEII y XhoI. La presencia del tipo genómico II había sido reportada en la Argentina una sola vez y, hasta el momento, no se han informado nuevos casos. Este estudio reveló una homogeneidad de los sitios BamHI, acorde con el número y el tamaño de los fragmentos. La presencia del fragmento BamHI #7 en los aislamientos argentinos analizados sugiere que éstos pertenecen al tipo salvaje (wild-type). Además, se caracterizó el principal dominio inmunogénico de la glicoproteína E de todas las cepas argentinas y se lo comparó con los de las cepas de referencia y con las secuencias disponibles en la base de datos GenBank. Los porcentajes de similitud obtenidos oscilaron entre 99 y 100%.The genomic characterization of Suid Herpesvirus 1 (SHV-1) isolates from Argentina was accomplished by restriction pattern analysis using the BamHI, BstEII and XhoI enzymes. Type II genome has been described only once in Argentina. This study revealed considerable homogeneity of BamHI endonuclease sites in all the strains analyzed, according to the number and size of the fragments. No deletion of BamHI fragment #7 among the Argentinean isolates suggests that these strains are wild-type. In addition, the main antigenic domain of glycoprotein E of all the Argentinean strains, as well as the reference strains and sequences available in the GenBank, were characterized. The similarity percent oscillated between 99 and 100%.Facultad de Ciencias Veterinaria
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Diagnóstico rápido del virus de la pseudorrabia en tejidos mediante el método de reacción en cadena de la polimerasa
2020Co-Authors: Echeverría, María Gabriela, Galosi, Cecilia Monica, Pidone, Claudio Luis, Pereyra N. B., Etcheverrigaray, Maria Elisa, Pecoraro, Marcelo Ricardo Ítalo, Nosetto, Edgardo OmarAbstract:In Argentina pseudorabies is an endemic disease. Routine diagnosis is made by virus isolation. It is a very long procedure to carry out and gives variable results depending on the quality of sample, hence the need for effective techniques, which are rapid and not dependent on the isolation of infectious virus. The polymerase chain reaction (PCR) technique has provided a sensitive, specific and rapid mean to detect DNA sequences. This study describes a PCR method for detection of pseudorabies virus sequences in swine tissues. In order to determine the presence of Suid Herpesvirus-1 DNA and antigens, 36 tissue samples collected from 19 dead pigs, with signs of pseudorabies infection, were examined by PCR, virus isolation and indirect immunofluorescence, respectively. Fifteen out of 19 pigs were positive at least for one tissue by PCR (15/19) while only three pseudorabies virus strains were isolated (3/19). All the amplified products were identified by digestion with Sa/l and hybridization. The method described herein circumvents tedious viral isolation and DNA purification and would be a valuable tool for rapid diagnosis, since it would take less than 5 h to reach an accurate result even in poorly preserved tissue samples.La pseudorrabia (enfermedad de Aujeszky) es endémica en la Argentina. El diagnóstico de rutina se realiza por aislamiento del virus, procedimiento lento y de resultados variables que dependen de la calidad de la muestra, por lo que es necesario una técnica efectiva y rápida como alternativa. La reacción en cadena de la polimerasa (PCR) permite detectar secuencias de ADN de forma rápida, específica y sensible. En este trabajo se describe un método de PCR para detectar secuencias de ADN del virus de la pseudorrabia a partir de tejidos. Se analizaron 36 órganos provenientes de 19 animales con sintomatología compatible con la enfermedad de Aujeszky, por PCR, aislamiento viral e inmunofluorescencia indirecta. Quince del total de casos analizados resultaron positivos por PCR, al menos para un tejido (15/19) mientras que, sólo tres cepas virales fueron aisladas (3/19). Todos los productos obtenidos resultaron específicos ya que fueron digeridos con Sa/I y reaccionaron frente a una sonda biotinilada. Consideramos que la técnica descripta es de mucha utilidad en el diagnóstico de enfermedad de Aujeszky, dado que no requiere purificar el ADN y permite obtener resultados en muestras aún no aptas para el aislamiento viral y en menos de 5 horas.Facultad de Ciencias Veterinaria
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Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs
'Elsevier BV', 2013Co-Authors: Serena, María Soledad, Metz, Germán Ernesto, Mórtola, Eduardo Carlos, Geisler Christoph, Corva, Santiago Gerardo, Larsen Alejandra, Jarvis, Donald L., Echeverría, María GabrielaAbstract:Suid Herpesvirus 1 (SHV-1) is the etiological agent of Aujeszky’s disease (AD), which affects swine herds worldwide and causes substantial economic losses due to animal mortality and lost productivity. In order to eradicate SHV-1, vaccination programs using viruses lacking the gene encoding glycoprotein E (gE) are ongoing in several countries. These eradication programs have generated a currently unmet demand for affordable and sensitive tests that can detect SHV-1 infection, yet distinguish between infected and vaccinated pigs. To meet this demand, we used the baculovirus-insect cell system to produce immunologically authentic full-length recombinant gE protein for use in a serum ELISA assay. As previous efforts to clone the gE gene had failed due to its extremely high GC-content (75% average), we used betaine as a PCR enhancer to facilitate amplification of the entire gE gene from the Argentinian CL15 strain of SHV-1. The cloned gE gene was expressed at high levels in recombinant baculovirus-infected insect cells and reacted strongly with sera from SHV-1 infected pigs. We used the recombinant gE protein to develop a local indirect ELISA test with sensitivity and specificity comparable to currently available commercial tests. Thus, recombinant gE produced in baculovirus-infected insect cells is a viable source of antigen for the detection of SHV-1 in ELISA tests. We also provide evidence supporting a potential application of this recombinant form of gE as a SHV-1 subunit vaccine.Fil: Serena, Maria Soledad. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Geisler, Christoph. University of Wyoming. Department of Molecular Biology; Estados UnidosFil: Metz, German Ernesto. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Corva, Santiago Gerardo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Mórtola, Eduardo Carlos. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Larsen, Alejandra. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Jarvis, Donald L.. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Echeverria, Maria Gabriela. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin
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Caracterización de cepas de campo de Herpesvirus suino 1 aisladas en Argentina
Asociación Argentina de Microbiología, 2010Co-Authors: Serena, María Soledad, Metz, Germán Ernesto, Martín Ocampos, Giselle Paula, Gambaro, Sabrina Eliana, Mortola Eduardo, Echeverría, María GabrielaAbstract:Caracterización de cepas de campo de Herpesvirus suino 1 aisladas en Argentina. Se caracterizaron cepas argentinas de Herpesvirus suino tipo 1 (HVS-1) mediante corte con las enzimas de restricción BamHI, BstEII y XhoI. La presencia del tipo genómico II había sido reportada en la Argentina una sola vez y, hasta el momento, no se han informado nuevos casos. Este estudio reveló una homogeneidad de los sitios BamHI, acorde con el número y el tamaño de los fragmentos. La presencia del fragmento BamHI #7 en los aislamientos argentinos analizados sugiere que éstos pertenecen al tipo salvaje (wild-type). Además, se caracterizó el principal dominio inmunogénico de la glicoproteína E de todas las cepas argentinas y se lo comparó con los de las cepas de referencia y con las secuencias disponibles en la base de datos GenBank. Los porcentajes de similitud obtenidos oscilaron entre 99 y 100%.The genomic characterization of Suid Herpesvirus 1 (SHV-1) isolates from Argentina was accomplished by restriction pattern analysis using the BamHI, BstEII and XhoI enzymes. Type II genome has been described only once in Argentina. This study revealed considerable homogeneity of BamHI endonuclease sites in all the strains analyzed, according to the number and size of the fragments. No deletion of BamHI fragment #7 among the Argentinean isolates suggests that these strains are wild-type. In addition, the main antigenic domain of glycoprotein E of all the Argentinean strains, as well as the reference strains and sequences available in the GenBank, were characterized. The similarity percent oscillated between 99 and 100%.Fil: Serena, Maria Soledad. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Metz, German Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; ArgentinaFil: Martin Ocampos, Giselle Paula. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; ArgentinaFil: Gambaro, Sabrina Eliana. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; ArgentinaFil: Mortola, Eduardo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Cátedra de Inmunología; ArgentinaFil: Echeverria, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentin
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Diagnóstico rápido del virus de la pseudorrabia en tejidos mediante el método de reacción en cadena de la polimerasa
Asociación Argentina de Microbiología, 2000Co-Authors: Echeverría, María Gabriela, Galosi, Cecilia Monica, Pidone, Claudio Luis, Pecoraro, Marcelo Ricardo, Pereyra N. B., Etcheverrigaray, Maria Elisa, Nosetto, Edgardo OmarAbstract:In Argentina pseudorabies is an endemic disease. Routine diagnosis is made by virus isolation. It is a very long procedure to carry out and gives variable results depending on the quality of sample, hence the need for effective techniques, which are rapid and not dependent on the isolation of infectious virus. The polymerase chain reaction (PCR) technique has provided a sensitive, specific and rapid mean to detect DNA sequences. This study describes a PCR method for detection of pseudorabies virus sequences in swine tissues. In order to determine the presence of Suid Herpesvirus-1 DNA and antigens, 36 tissue samples collected from 19 dead pigs, with signs of pseudorabies infection, were examined by PCR, virus isolation and indirect immunofluorescence, respectively. Fifteen out of 19 pigs were positive at least for one tissue by PCR (15/19) while only three pseudorabies virus strains were isolated (3/19). All the amplified products were identified by digestion with Sa/l and hybridization. The method described herein circumvents tedious viral isolation and DNA purification and would be a valuable tool for rapid diagnosis, since it would take less than 5 h to reach an accurate result even in poorly preserved tissue samples.Fil: Echeverria, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Area de Virología; ArgentinaFil: Pecoraro, Marcelo Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Area de Virología; ArgentinaFil: Pereyra, N. B.. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Area de Virología; ArgentinaFil: Pidone, Claudio Luis. Universidad Nacional de Rosario. Facultad de Ciencias Veterinarias; ArgentinaFil: Galosi, Cecilia Monica. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Area de Virología; ArgentinaFil: Etcheverrigaray, Maria Elisa. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Area de Virología; ArgentinaFil: Nosetto, Edgardo Omar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Area de Virología; Argentin
Laurids Siig Christensen - One of the best experts on this subject based on the ideXlab platform.
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the population biology of Suid Herpesvirus 1
Acta Pathologica Microbiologica et Immunologica Scandinavica, 1995Co-Authors: Laurids Siig ChristensenAbstract:Suid Herpesvirus 1 (SHV-1) is the causative agent of Aujeszky's disease (AD, pseudorabies), known worldwide as a major economical threat to pig farming. Measures are taken in many countries to control or eradicate the disease and considerable effort therefore has been focused on the elucidation of the epizootiology of the infection. These studies were greatly facilitated by the possibilities, first reported in the early eighties, to identify SHV-1 strains by means of restriction fragment pattern (RFP) analysis. In the present thesis some molecular biological aspects of SHV-1 are reviewed. In addition, studies are reviewed focusing on three topics: (i) methodological aspects of molecular strain identification, (ii) various epizootiological features of the SHV-1 infection, and (iii) the population dynamics of coexisting virus particles comprising a field strain, tentatively defined as a transmissible entity. A systematization of the European isolates of SHV-1 was elaborated based on molecular characterization of various genome types and an evolutionary tree for some of the distinct types was suggested. For some of the types geographical niches could be identified indicating that the intertypic differences had been stable for decades. By the characterization of 5-10 isolates from each herd, in which a SHV-1 strain had been newly introduced, strain inherent non-intertypic genomic variations consistently could be demonstrated. Some strains appeared to consist of fairly homogeneous pools of genomic variants, while other strains appeared highly heterogeneous. Some exhibited hypervariable regions in the genome. The pool of genomic variants present in a strain was found to be a highly specific and most often a conservative characteristic of a strain. Yet, fluctuations in the proportions of subpopulations occasionally were seen. Thus, while one isolate from an outbreak might be a poor representative of the pool of variants comprising the causative strain, the analysis of 5-10 isolates from each outbreak might taken together provide the basis of an extremely fine resolving potential. Outbreaks of AD during the finishing stage of the Danish eradication campaign were subjected to intensive molecular epizootiological studies. The analysis of representative older isolates of SHV-1 revealed that only type III was present in Denmark prior to 1985. In 1985 type IIa isolates emerged in a border area, and since 1986 type IIp and IIa were the only types identified in Denmark. Severe epizootics have been recorded in border areas in Denmark since the winter of 1986/87.(ABSTRACT TRUNCATED AT 400 WORDS)
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High frequency intergenomic recombination of Suid Herpesvirus 1 (SHV-1, Aujeszky's disease virus)
Archives of Virology, 1993Co-Authors: Laurids Siig Christensen, B. LomnicziAbstract:Examples are given of observations made with field isolates of Suid Herpesvirus 1 (SHV-1) which indicate that intergenomic recombination is a common phenomenon associated with the virus. This was further confirmed by experimental co-infection of a pig with 2 virus strains with different, stable and easily identifiable genomic markers, followed by natural transmission to a group of contact pigs. A variety of recombinants was subsequently isolated, while none of the parental strains were re-isolated from any of the pigs. It is suggested that co-invasion of cells and recombination between viral genomes play a role in the life cycle of the virus.
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characterization of field isolates of Suid Herpesvirus 1 aujeszky s disease virus as derivatives of attenuated vaccine strains
Archives of Virology, 1992Co-Authors: Laurids Siig Christensen, I Medveczky, Bertel Strandbygaard, Z PejsakAbstract:Field isolates of Suid Herpesvirus 1 (Aujeszky's disease virus) from Poland and Hungary were identified by restriction fragment pattern analysis as derivatives of attenuated vaccine strains. The Polish isolates were found to be related to the BUK-TK-900 strain (Suivac A) which is widely used as a live vaccine in Poland, and the Hungarian isolates were related to the Bartha K-61 vaccine strain widely used in Hungary. Pigs experimentally infected with derivatives of BUK-TK-900 or BUK-TK-900 itself were found to develop gI-antibodies, while pigs infected with derivatives of Bartha K-61 showed a gI-negative response.
Maria Soledad Serena - One of the best experts on this subject based on the ideXlab platform.
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expression and purification of Suid Herpesvirus 1 glycoprotein e in the baculovirus system and its use to diagnose aujeszky s disease in infected pigs
Protein Expression and Purification, 2013Co-Authors: Maria Soledad Serena, German Ernesto Metz, Eduardo Carlos Mortola, Christoph Geisler, Santiago Corva, Alejandra Larsen, Donald L Jarvis, Maria Gabriela EcheverriaAbstract:Suid Herpesvirus 1 (SHV-1) is the etiological agent of Aujeszky’s disease (AD), which affects swine herds worldwide and causes substantial economic losses due to animal mortality and lost productivity. In order to eradicate SHV-1, vaccination programs using viruses lacking the gene encoding glycoprotein E (gE) are ongoing in several countries. These eradication programs have generated a currently unmet demand for affordable and sensitive tests that can detect SHV-1 infection, yet distinguish between infected and vaccinated pigs. To meet this demand, we used the baculovirus-insect cell system to produce immunologically authentic full-length recombinant gE protein for use in a serum ELISA assay. As previous efforts to clone the gE gene had failed due to its extremely high GC-content (75% average), we used betaine as a PCR enhancer to facilitate amplification of the entire gE gene from the Argentinian CL15 strain of SHV-1. The cloned gE gene was expressed at high levels in recombinant baculovirus-infected insect cells and reacted strongly with sera from SHV-1 infected pigs. We used the recombinant gE protein to develop a local indirect ELISA test with sensitivity and specificity comparable to currently available commercial tests. Thus, recombinant gE produced in baculovirus-infected insect cells is a viable source of antigen for the detection of SHV-1 in ELISA tests. We also provide evidence supporting a potential application of this recombinant form of gE as a SHV-1 subunit vaccine.
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phylogenetic analysis of Suid Herpesvirus 1 isolates from argentina
Veterinary Microbiology, 2011Co-Authors: Maria Soledad Serena, German Ernesto Metz, Eduardo Carlos Mortola, Maria Gabriela EcheverriaAbstract:Argentinean Suid Herpesvirus 1 isolates were compared with reference strains and sequences available at GenBank and phylogenetically analyzed. A short fragment of the gE gene of the immunodominant epitopes was used for preliminary grouping of isolates by phylogenetic analysis. The analysis of the partial gC gene provided more precise genetic typing and segregation into the main genotypes I and II. Results confirmed that the Argentinean genotype I isolates predominate in our country. The topology of the partial gC gene was similar to that previously reported. The Argentinean type I isolates belonged to one cluster and grouped together with NIA-3 and American and Brazilian genotype I strains. However, the results obtained by the algorithms allow inferring that the Yamagata S-81 and Mer (genotype II) strains are grouped together.
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characterization of Suid Herpesvirus 1 field isolates from argentina
Revista Argentina De Microbiologia, 2010Co-Authors: Maria Soledad Serena, German Ernesto Metz, Eduardo Carlos Mortola, G Martin P Ocampos, S E Gambaro, Maria Gabriela EcheverriaAbstract:The genomic characterization of Suid Herpesvirus 1 (SHV-1) isolates from Argentina was accomplished by restriction pattern analysis using the BamHI, BstEII and XhoI enzymes. Type II genome has been described only once in Argen- tina. This study revealed considerable homogeneity of BamHI endonuclease sites in all the strains analyzed, accord- ing to the number and size of the fragments. No deletion of BamHI fragment #7 among the Argentinean isolates suggests that these strains are wild-type. In addition, the main antigenic domain of glycoprotein E of all the Argentinean strains, as well as the reference strains and sequences available in the GenBank, were characterized. The similarity percent oscillated between 99 and 100%.
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Characterization of Suid Herpesvirus 1 field isolates from Argentina Caracterización de cepas de campo de Herpesvirus suino 1 aisladas en Argentina
Elsevier España S.L.U., 2010Co-Authors: Maria Soledad Serena, German Ernesto Metz, Eduardo Carlos Mortola, G Martin P Ocampos, S E Gambaro, Maria Gabriela EcheverriaAbstract:The genomic characterization of Suid Herpesvirus 1 (SHV-1) isolates from Argentina was accomplished by restriction pattern analysis using the BamHI, BstEII and XhoI enzymes. Type II genome has been described only once in Argentina. This study revealed considerable homogeneity of BamHI endonuclease sites in all the strains analyzed, according to the number and size of the fragments. No deletion of BamHI fragment #7 among the Argentinean isolates suggests that these strains are wild-type. In addition, the main antigenic domain of glycoprotein E of all the Argentinean strains, as well as the reference strains and sequences available in the GenBank, were characterized. The similarity percent oscillated between 99 and 100%.Se caracterizaron cepas argentinas de Herpesvirus suino tipo 1 (HVS-1) mediante corte con las enzimas de restricción BamHI, BstEII y XhoI. La presencia del tipo genómico II había sido reportada en la Argentina una sola vez y, hasta el momento, no se han informado nuevos casos. Este estudio reveló una homogeneidad de los sitios BamHI, acorde con el número y el tamaño de los fragmentos. La presencia del fragmento BamHI #7 en los aislamientos argentinos analizados sugiere que éstos pertenecen al tipo salvaje (wild-type). Además, se caracterizó el principal dominio inmunogénico de la glicoproteína E de todas las cepas argentinas y se lo comparó con los de las cepas de referencia y con las secuencias disponibles en la base de datos GenBank. Los porcentajes de similitud obtenidos oscilaron entre 99 y 100%
German Ernesto Metz - One of the best experts on this subject based on the ideXlab platform.
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expression and purification of Suid Herpesvirus 1 glycoprotein e in the baculovirus system and its use to diagnose aujeszky s disease in infected pigs
Protein Expression and Purification, 2013Co-Authors: Maria Soledad Serena, German Ernesto Metz, Eduardo Carlos Mortola, Christoph Geisler, Santiago Corva, Alejandra Larsen, Donald L Jarvis, Maria Gabriela EcheverriaAbstract:Suid Herpesvirus 1 (SHV-1) is the etiological agent of Aujeszky’s disease (AD), which affects swine herds worldwide and causes substantial economic losses due to animal mortality and lost productivity. In order to eradicate SHV-1, vaccination programs using viruses lacking the gene encoding glycoprotein E (gE) are ongoing in several countries. These eradication programs have generated a currently unmet demand for affordable and sensitive tests that can detect SHV-1 infection, yet distinguish between infected and vaccinated pigs. To meet this demand, we used the baculovirus-insect cell system to produce immunologically authentic full-length recombinant gE protein for use in a serum ELISA assay. As previous efforts to clone the gE gene had failed due to its extremely high GC-content (75% average), we used betaine as a PCR enhancer to facilitate amplification of the entire gE gene from the Argentinian CL15 strain of SHV-1. The cloned gE gene was expressed at high levels in recombinant baculovirus-infected insect cells and reacted strongly with sera from SHV-1 infected pigs. We used the recombinant gE protein to develop a local indirect ELISA test with sensitivity and specificity comparable to currently available commercial tests. Thus, recombinant gE produced in baculovirus-infected insect cells is a viable source of antigen for the detection of SHV-1 in ELISA tests. We also provide evidence supporting a potential application of this recombinant form of gE as a SHV-1 subunit vaccine.
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phylogenetic analysis of Suid Herpesvirus 1 isolates from argentina
Veterinary Microbiology, 2011Co-Authors: Maria Soledad Serena, German Ernesto Metz, Eduardo Carlos Mortola, Maria Gabriela EcheverriaAbstract:Argentinean Suid Herpesvirus 1 isolates were compared with reference strains and sequences available at GenBank and phylogenetically analyzed. A short fragment of the gE gene of the immunodominant epitopes was used for preliminary grouping of isolates by phylogenetic analysis. The analysis of the partial gC gene provided more precise genetic typing and segregation into the main genotypes I and II. Results confirmed that the Argentinean genotype I isolates predominate in our country. The topology of the partial gC gene was similar to that previously reported. The Argentinean type I isolates belonged to one cluster and grouped together with NIA-3 and American and Brazilian genotype I strains. However, the results obtained by the algorithms allow inferring that the Yamagata S-81 and Mer (genotype II) strains are grouped together.
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characterization of Suid Herpesvirus 1 field isolates from argentina
Revista Argentina De Microbiologia, 2010Co-Authors: Maria Soledad Serena, German Ernesto Metz, Eduardo Carlos Mortola, G Martin P Ocampos, S E Gambaro, Maria Gabriela EcheverriaAbstract:The genomic characterization of Suid Herpesvirus 1 (SHV-1) isolates from Argentina was accomplished by restriction pattern analysis using the BamHI, BstEII and XhoI enzymes. Type II genome has been described only once in Argen- tina. This study revealed considerable homogeneity of BamHI endonuclease sites in all the strains analyzed, accord- ing to the number and size of the fragments. No deletion of BamHI fragment #7 among the Argentinean isolates suggests that these strains are wild-type. In addition, the main antigenic domain of glycoprotein E of all the Argentinean strains, as well as the reference strains and sequences available in the GenBank, were characterized. The similarity percent oscillated between 99 and 100%.
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Characterization of Suid Herpesvirus 1 field isolates from Argentina Caracterización de cepas de campo de Herpesvirus suino 1 aisladas en Argentina
Elsevier España S.L.U., 2010Co-Authors: Maria Soledad Serena, German Ernesto Metz, Eduardo Carlos Mortola, G Martin P Ocampos, S E Gambaro, Maria Gabriela EcheverriaAbstract:The genomic characterization of Suid Herpesvirus 1 (SHV-1) isolates from Argentina was accomplished by restriction pattern analysis using the BamHI, BstEII and XhoI enzymes. Type II genome has been described only once in Argentina. This study revealed considerable homogeneity of BamHI endonuclease sites in all the strains analyzed, according to the number and size of the fragments. No deletion of BamHI fragment #7 among the Argentinean isolates suggests that these strains are wild-type. In addition, the main antigenic domain of glycoprotein E of all the Argentinean strains, as well as the reference strains and sequences available in the GenBank, were characterized. The similarity percent oscillated between 99 and 100%.Se caracterizaron cepas argentinas de Herpesvirus suino tipo 1 (HVS-1) mediante corte con las enzimas de restricción BamHI, BstEII y XhoI. La presencia del tipo genómico II había sido reportada en la Argentina una sola vez y, hasta el momento, no se han informado nuevos casos. Este estudio reveló una homogeneidad de los sitios BamHI, acorde con el número y el tamaño de los fragmentos. La presencia del fragmento BamHI #7 en los aislamientos argentinos analizados sugiere que éstos pertenecen al tipo salvaje (wild-type). Además, se caracterizó el principal dominio inmunogénico de la glicoproteína E de todas las cepas argentinas y se lo comparó con los de las cepas de referencia y con las secuencias disponibles en la base de datos GenBank. Los porcentajes de similitud obtenidos oscilaron entre 99 y 100%
