Home
 /  Seminar  /  Seminar 820
Seminar 820  
Date:2012-12-12
Update:2014-01-09

Seminar:  820 

1

Speaker(s)

Shu-Chun Chiu

Topic

Study on pathogenesis of porcine teschovirus and development of its vaccine

Abstract

The Teschen-Talfan disease is also known as Teschen disease, poliomyelitis suum or porcine enterovirus encephalomyelitis. The disease was initially observed in Czechoslovakia in 1929, and is caused by certain strains of porcine teschoviruses (PTV) classified to the family Picornaviridae. The most severe nervous lesions occurred in the brainstem from the hypothalamus through the medulla, and decreased in intensity and diffuseness down the spinal cord. In enterovirus encephalomyelitis of pigs in Japan in 2002, no PTV was isolated from the cerebrum or spinal cord, whereas PTV was isolated from a clinically normal piglet, suggesting that many infections with this virus are asymptomatic and the epidemiology and mechanism of the neurovirulence of PTV still needs to be determined. In Taiwan, PTV-1s were isolated and identified from clinical cases submitted to Animal Health Research Institute in years 2000, 2003, 2004, 2005 and 2006, respectively.  Based on these results, it is suggested that teschovirus infection may have occurred widely in pig population of Taiwan. In this study, experiments were done to realize the pathogenesis of porcine teschovirus. And then, we want to understand the localization of PTV-1 by RT-PCR, IHC methodology and the potential causes of background, it is important to be aware of the various pathways of DNA synthesis that may be operative inside an intact cell.

2

Speaker(s)

T.S. Huang

Topic

Development of a contagious pustular dermatitis attenuated live vaccine and its antibody detection kit

Abstract

Contagious Pustular Dermatitis, which is also called contagious pustular stomatitis, contagious ecthyma, soremouth, infectious labial dermatitis, scabby mouth and Orf, etc. is a highly infectious viral disease of sheep and goats, characterized by the development of pustular and scabby lesions on the muzzle and lips. Three and two Orf field strains isolated in 2011 and 2012 have been propagated in GT and CEF cells for 43 and 12 passages, respectively. Two batches of the trial vaccines, 10,000 doses for each, were manufactured in chicken embryo fibroblast (CEF) cell monolayer inoculated with the 43rd cell passage and attenuated Orf-1 seed virus. Each of 3-month-old goats was intracutaneously inoculated with 20 doses of the trial vaccine, respectively. They showed no systemic syndromes in clinical or specific Orf lesions in necropsy except the redness and swelling for 5-6 days at the inoculation sites. It indicated the goats got successfully take and the attenuated Orf-1 seed virus was highly safe. Furthermore, the B2L genes of 5 isolates, 1,137 bp in length, were amplified with B2L gene specific primers by PCR and sequenced. Comparing to the Taiping strain, the B2L genes of 5 isolates showed only a nucleotide difference in nucleotide sequence, but it showed 100% similarity in amino acid sequence. The recombinant B2L proteins were successfully expressed with both eukaryotic and prokaryotic expression system and will be used for the following ELISA kit and subunit vaccine tests.

3

Speaker(s)

Chu-Hsiang Pan

Topic

Use of attenuated Salmonella Typhimurium as carrier to express viral antigen. Report on the validation of the detection of H3N2 variant influenza A virus using RT-PCR

Abstract

An auto-display plasmid for bacterial surface expression of recombinant protein was constructed and transformed into the attenuated Salmonella typhimurium strain for auto expression of ORF2 protein of porcine circle virus type 2 (PCV2). The transformed Salmonella typhimurium Mice were fed with Salmonella typhimurium to evaluate the possibility of bacteria as vector for vaccine delivery by detection the antibodies against PCV2. First,ORF2 gene was subcloned into the plasmids of pRSET-B.  Then the gene of adhesin involved in diffuse adherence (AIDA) was subcloned into the plasmid of ORF2-pRSETB and constructed a surface displaying system. To construct an auto-display plasmid for expression of recombinant protein on surface of  Salmonella strain, three gene fragments containing fen promoter, terminator and T7 RNA polymerase were subcloned into the plasmid individually. Then the plasmid was transformed into the Salmonella typhimurium strain, Top10 and DH5α E. coli strains respectively. Results showed that the recombinant proteins of ORF2 could be expressed by these three transformed bacteria and it could be detected by Western blot.

We had been to College of Veterinary Medicine at Kansas State University, USA during from November 16 to November 25, 2012

, the purpose of this study was detect antibodies against influenza virus A/ Taiwan? /2009/ (H1N1) with ELISA, and detect influenza virus A/ Taiwan? /?/ (H3N2) with RT-PCR. to evaluate an ELISA method for detection of antibodies against pandemic 2009 Influenza A (H1N1) virus and to evaluate a RT-PCR method for detection of H3N2 variant (H3N2v) influenza, virus

Three primer sets were designed based on hemagglutinin (HA), neuraminidase (NA) and matrix (M) gene of H3N2v virus. One-step RT-PCR was used for rapid differential detection of H3N2v virus among the 13 samples of H3N2v, H1N2v, human seasonal influenza viruses,traditional swine influenza viruses. Results showed that RT-PCR method could be used for rapid and sensitivity differential detection of H3N2v viruses.