Detection of RD-114 virus by a reverse transcriptase assay based on product enhancement

• Published in: Biologicals Vol. 40; no. 6; pp. 421 - 425
• Main Authors: Narushima, Rie, Tamura, Naoya, Iida, Masayuki, Saito, Akito
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01-11-2012
• Subjects: Animals; Base Sequence; Canine live vaccines; Cats; Cell culture; Cell Line; Corticotropin-releasing hormone; DNA Primers; dogs; Endogenous retrovirus; Endogenous Retroviruses - enzymology; Endogenous Retroviruses - metabolism; genome; Genomes; HEK293 Cells; Humans; Kidney; kidneys; live vaccines; PERT assay; RD-114 virus; Real-Time Polymerase Chain Reaction; Retroviridae; Retrovirus; Reverse transcriptase; RNA-directed DNA polymerase; RNA-Directed DNA Polymerase - metabolism; Vaccines; viruses; PERT assay; Endogenous retrovirus; Reverse transcriptase; RD-114 virus; Canine live vaccines
• ISSN: 1045-1056; 1095-8320
• DOI: 10.1016/j.biologicals.2012.09.002

RD-114 virus is a feline endogenous retrovirus that exists in the genome of all cats. It can be assumed that feline and canine live vaccines manufactured by culturing cells of feline origin are contaminated with the virus. The current study is attempted to develop a product enhanced reverse transcriptase (PERT) assay to detect replication-competent RD-114 virus. Since culture supernatants of Crandell-Rees feline kidney (CRFK) cells do not have detectable reverse transcriptase activity in case of do not passage on other cell lines, these results raise the possibility that RD-114 virus grow efficiently in other retrovirus producing cell lines. For the PERT assay, RD-114 virus isolated from CRFK cells may need to be passaged more than four times on 293T cells to be expressed at appreciable levels. The PERT assay described here provides an accurate method for determining the presence of RD-114 in culture supernatants and will help monitor live vaccines produced in endogenous virus producing cell lines.