Improve the potency assay for BCG vaccine and HCV genotyping assay by novel biotechniques
Date Issued
2014
Date
2014
Author(s)
Yang, Yi-Chen
Abstract
Most of the traditional assay used for biologics evaluation & clinical diagnosis, such as direct culturing and counting Colony-Forming Unit (CFU) for potency assay of live attenuated vaccine or direct sequencing for virus genotyping are labor-intensive or time-consuming, and depend upon visual read-out of the image, which may cause different results due to different operators. Since the novel techniques for rapid or multiplexing detection are getting more popular, we therefore tried to improve the potency assay for biologics evaluation and the virus genotyping detection assay using the novel biotechniques.
The Bacille Calmette-Guerin (BCG) vaccine is a live attenuated vaccine prepared from a strain of Mycobacterium bovis. The conventional method to determine the potency of the BCG vaccine is to count the number of colony forming units (CFU). However, M. bovis is a slow growing organism and it takes at least four weeks incubation for the potency assay. The results of the potency assay could also vary due to manual counting. Therefore, we developed a rapid and reliable method to determine the potency of M. bovis BCG. The total handing time of the assay is only 4 hours and the results showed a very good agreement with the results from conventional CFU method.
In addition, there are more than six different genotypes of hepatitis C virus (HCV) around the world and they play an important role in the treatment of Hepatitis C patients. However, most of the current genotyping methods are unable to detect mixed genotypes from two or more HCV infections. In order to detect different HCV genotypes in a sample, we applied suspension bead array technology to identify HCV genotypes simultaneously. The assay is capable to detect two different HCV genotypes within a sample and identify HCV in HCV/HIV mixed samples. The features of rapid and reliable make this assay became a potential genotyping method for HCV genotyping in the future.
Our studies demonstrate that the novel biotechniques can improve the potency assay for biologics evaluation and the virus genotyping assay. Therefore, improvement strategies utilizing novel biotechniques for virus genotyping and biologics’ potency measurement could be constantly applied in the future.
Subjects
流式細胞儀
卡介苗
效價試驗
懸浮微珠偵測系統
Ｃ型肝炎病毒
基因分型方法
SDGs
Type
thesis
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