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Seminar 874  
Date:2017-06-20
Update:2017-11-07

  

Seminar:  874  

1

Speaker(s)

Chieh-Hao Wu

Topic

A summary of transmission electron microscopy analyses in 2016

Abstract

Transmission electron microscopy (TEM) is a powerful tool in identifying cells and elucidating the ultrastructure of cellular components especially in the fields of virology and microbiology. We have used TEM primarily to examine potential pathogens in terrestrial and aquatic animals such that they can be detected in pathogen microoganisms earlier. The results can thus be used to further guide direction of specific tests for rapid disease diagnosis.

A total of 286 samples were collected for TEM in 2016. In addition to fourteen samples processed with the ultramicrotome, 272 samples encompassing 87 samples from poultry (31.9%), 73 samples from herbivores (26.8%), 72 samples from aquatic animals (26.4%), 24 samples from swine (8.9%), and 16 samples from other sample types (6%) were examined by negative staining for pathogen observation. In 119 of those specimens, we were able to positively identify specific virons and bacteria.

2

Speaker(s)

Chu-Hsiang Pan

Topic

Development and application of an ELISA test for the detection of E2 antibodies against classical swine fever virus

Abstract

The Lapinized Philippines Coronel (LPC) vaccine is widely used in Taiwan to prevent classical swine fever (CSF). Furthermore, classical swine fever antibody titers are frequently used to assess immune response, vaccine efficacy, as well as enabling one to adjust the appropriate vaccine program by means of detecting the maternal antibody titer in piglets. The serum neutralization test (SNT) is considered as the “gold standard” for the detection of CSF antibody titers. However, traditional SNT is a time-consuming (3 days) and labor intesinve procedure, involving cell culture, serum-virus neutralization, fluorescent-antibody staining, and visual interpretation under fluorescence microscopy. On the other hand, the detection of CSF antibodies using ELISA is a relatively straight forward procedure and easily processed in order to enhance the efficiency of CSF antibody detection, we cooperated with the Kaohsiung Medical University using a mammalian cell line that produces high levels of the E2 CSF virus glycoprotein. We also developed indirect and blocking ELISA assays for the detection of antibodies against the CSF virus. A total of 400 serum samples collected from pig farms were tested by homemade indirect and blocking ELISA kits. The results were compared with the SNT titers for the purpose of evaluating the feasibility of the ELISA method for serological monitoring. Two homemade ELISA kits and two commercial ELISA kits were tested and analyzed. The results demonstrated that the correlation coefficient between SNT titer and the optical density (OD) values of the homemade indirect and blocking ELISA kits were 0.85 and 0.86, respectively. In addition, the correlation coefficient between the homemade ELISA kits and the commercially available indirect and blocking ELISA kits were 0.88 and 0.94, respectively. These results indicate that the level of anti-E2 antibodies which were detected by using the indirect and blocking ELISA tests were correlated with the antibody titers of SNT.