Replication-Competent Adenovirus Assay
Replication-competent adenovirus detection is necessary for adenovirus vector safety evaluation. Armed with an advanced gene therapy platform, Creative Biolabs offers rapid and professional assays for the replication-competent adenovirus detection to ensure the smooth progress of follow-up research.
Introduction of Adenovirus Vectors
Adenoviruses are a class of double-stranded DNA viruses with an icosahedral nucleocapsid and none lipid envelope. A diversity of adenoviruses have been identified, and over 50 different types of adenoviruses have been found to cause varying degrees of infections in humans. Currently, adenovirus is a potential and excellent viral vector for gene therapy since that (i) high-efficiency in infection of both replicating and non-replicating cells; (ii) ability to transduce a broad range of hosts; (iii) encoding protein expression without integration into the genome of the host cell. One of the most thoroughly studied and extensively utilized adenoviral vectors is adenovirus serotype 5.
Usually, there are two categories: replication-defective adenoviruses vectors and replication-competent adenoviruses vectors. A replication-competent adenovirus is produced by introducing an exogenous gene to replace the deleted E3 region (a non-essential for viral replication conservative sequence), or replication-defective adenoviruses recombining with host sequences. Replication-competent adenovirus has made great achievements in the fight against cancer as an oncolytic vector.
Figure 1. Schematic of the adenovirus genome and vectors (replication-defective and replication-competent adenovirus vectors). (Doronin, 2000)
Replication-Competent Adenovirus Assay
Replication-competent adenovirus has been extensively studied and applied in antineoplastic gene therapy, but there are also unsafe factors due to their ability to replicate. More importantly, replication-competent adenovirus increases with each successive amplification, and the risk of infection increases accordingly. Thus, sensitive assays for the detection of replication-competent recombinant adenoviruses are necessary to adenovirus vector characterization. We provide professional replication-competent adenovirus detecting assays, including but not limited to:
- Cytopathic effect detection assay: observation and analysis of the cytopathic effect following the infection and amplification of recombinant adenovirus vectors.
- Rapid determination the replication-competent adenovirus by the combination of microcarrier cell culture and polymerase chain reaction (PCR): incubation of adenoviral vectors on A549 cells in a microcarrier cell culture system followed by detection of amplified replication-competent adenoviruses by E1-specific quantitative PCR (Schalk, 2007).
- Other detection assays: combination replication-competent adenovirus amplification with immunocytochemistry staining, cytopathic effect evaluation followed by differential amplification of adenoviral vector, etc.
Services
As a leading company in viral vector construction, Creative Biolabs provides a series of replication-competent adenovirus detection services. Moreover, our scientists, specialized in gene therapy, has successfully established a comprehensive and perfect gene therapy services system aiming to offer rapid and one-stop viral vector-based gene therapy services to customers all over the world.
Please directly contact us for more detailed information.
References
- Doronin, K.; et al. (2000). Tumor-Specific, Replication-Competent Adenovirus Vectors Overexpressing the Adenovirus Death Protein. Journal of Virology. 74(13): 6147-6155.
- Schalk, J.A.; et al. (2007). A rapid and sensitive assay for detection of replication-competent adenoviruses by a combination of microcarrier cell culture and quantitative PCR. Journal of Virological Methods. 145(2): 89-95.
