Analysis of the virus propagation profile of 14 dengue virus isolates in Aedes albopictus C6/36 cells
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(2020) 13:481 Hitakarun et al. BMC Res Notes https://doi.org/10.1186/s13104-020-05325-6
Open Access
RESEARCH NOTE
Analysis of the virus propagation profile of 14 dengue virus isolates in Aedes albopictus C6/36 cells Atitaya Hitakarun, Suwipa Ramphan, Nitwara Wikan and Duncan R. Smith*
Abstract Objective: The mosquito transmitted RNA virus dengue virus (DENV) shows significant variation as a consequence of the lack of proofreading activity of the RNA-dependent RNA polymerase that synthesizes new virus genomes. How this variation affects DENV replication, and how this in turn impacts drug development remains largely unknown. Given the technical limitations in working with large numbers of isolates few studies have sought to investigate this area. This study used a panel of 14 DENV isolates of different serotypes and origins to determine how much virus replication in Aedes albopictus C6/36 cells was affected by DENV variability. Results: The results showed that there was considerable variation, with peak titers ranging from 6Log10 to 8Log10, and maximum titer being reached from day 3 to day 9 post infection. While strains from DENV 1 and 4 serotypes showed considerable uniformity, DENV 2 and 3 strains showed much greater variation. Overall, these results show that serotype specific strain variation can have a significant impact on DENV replication, suggesting that studies either investigating DENV pathogenesis or developing drug therapeutics should consider the contribution of DENV variability. Keywords: Dengue virus, Genetic diversity, Virus production profile Introduction Dengue virus (DENV) is an enveloped virus and composed of a single-stranded RNA genome encoding for three structural proteins (capsid (C), pre-membrane (prM), and envelope (E)) and seven non-structural (NS) proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5) [1]. The NS5 protein is the largest and most conserved protein and encodes a methyltransferase in the N-terminal domain and a RNA-dependent RNA polymerase (RdRP) in the C-terminal domain, with the domains being separated by a short flexible linker [2]. As *Correspondence: [email protected]; duncan.smi@mahidol. ac.th Molecular Pathology Laboratory, Institute of Molecular Biosciences, Mahidol University, Salaya Campus, 25/25 Phuttamonthon Sai 4, Salaya, Nakhon Pathom 73170, Thailand
with other RNA viruses, the DENV RdRP lacks proofreading activities, and as such, DENV has considerable variation, with each serotype consisting of multiple lineages each of which consists of many different strains [3]. In a previous study we evaluated the replication kinetics of four laboratory adapted DENVs in Aedes (Ae.) albopictus C6/36 cells, investigating one strain for each serotype [4]. The results of that study showed that there was very little variation in virus production kinetics over an intermediate time period (up to 15 days post infection), and that the titers of the four DENVs differed by less than 1 Log10. Given that we have collected a relatively large panel of low passage DENV
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