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- Genetic changes in plaque-purified varicella vaccine strain Suduvax during in vitro propagation in cell culture
- Hye Rim Hwang , Se Hwan Kang , Chan Hee Lee
- J. Microbiol. 2021;59(7):702-707. Published online June 1, 2021
- DOI: https://doi.org/10.1007/s12275-021-1062-3
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Abstract
- Infection by varicella-zoster virus (VZV) can be prevented by using live attenuated vaccines. VZV vaccine strains are known to evolve rapidly in vivo, however, their genetic and biological effects are not known. In this study, the plaque-purified vaccine strain Suduvax (PPS) was used to understand the genetic changes that occur during the process of propagation in in vitro cell culture. Full genome sequences of three different passages (p4, p30, and p60) of PPS were determined and compared for genetic changes. Mutations were found at 59 positions. The number of genetically polymorphic sites (GPS) and the average of minor allele frequency (MAF) at GPSs were not significantly altered after passaging in cell culture up to p60. The number of variant nucleotide positions (VNPs), wherein GPS was found in at least one passage of PPS, was 149. Overall, MAF changed by less than 5% at 52 VNPs, increased by more than 5% at 42 VNPs, and decreased by more than 5% at 55 VNPs in p60, compared with that seen in p4. More complicated patterns of changes in MAF were observed when genetic polymorphism at 149 VNPs was analyzed among the three passages. However, MAF decreased and mixed genotypes became unequivocally fixed to vaccine type in 23 vaccine-specific positions in higher passages of PPS. Plaque-purified Suduvax appeared to adapt to better replication during in vitro cell culture. Further studies with other vaccine strains and in vivo studies will help to understand the evolution of the VZV vaccine.
Research Support, Non-U.S. Gov't
- Quantification of Toxigenic Microcystis spp. in Freshwaters by Quantitative Real-time PCR Based on the Microcystin Synthetase A Gene
- Kyoung-Hee Oh , Dong-Hwan Jeong , Young-Cheol Cho
- J. Microbiol. 2013;51(1):18-24. Published online March 2, 2013
- DOI: https://doi.org/10.1007/s12275-013-2354-z
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- 14 Scopus
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Abstract
- A method to estimate the abundance of toxigenic Microcystis in environmental samples by using quantitative real-time PCR was developed and optimized. The basis of this method is the amplification of a highly conserved region of the mcyA gene within the microcystin synthetase gene cluster. Using this method, the average copy number of mcyA gene per cell in toxigenic Microcystis strains was estimated. The molecular markers and method developed in this study can be used to monitor toxigenic strains of Microcystis in Korean freshwaters, in which harmful cyanobacterial blooms are routinely found.
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