Detection and significance of retroviral reverse transcriptase present in cell cultures

Kutle, Leonida (2014) Detection and significance of retroviral reverse transcriptase present in cell cultures. Doctoral thesis, Faculty of Science > Department of Biology.

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Abstract

The enzyme reverse transcriptase (RT) is a part of all retroviruses, and the presence of retroviruses in biological samples can be detected by measuring RT's activity. Product Enhanced Reverse Transcriptase (PERT) test is a very sensitive assay for detection of low concentrations of RT in a sample. To make the assay regulatory acceptable, a standard like avian myeloblastosis virus RT should be included. All of the samples originated from chicken substrate show RT activity. RT activity is detected also in samples from diploid and continuous cell cultures, and in the cell culture media with foetal bovine serum. Detected activity originates from non-infectious retroviral particles, as no rise of the RT activity in indicator cell cultures is detected, nor it negatively affects mice and guinea-pigs in abnormal toxicity test. The effect of various operating conditions during the production of viral vaccines on RT activity was tested. Chromatography on monolithic columns enables concentration of endogenous retroviruses as well as RT molecules.

Item Type: Thesis (Doctoral thesis)
Keywords: endogenous retroviruses, PERT assay, SPF eggs, live viral vaccines, chromatography, monolithic column, CIM-disks
Supervisor: Mažuran, Renata
Date: 2014
Number of Pages: 133
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Grozdana Sirotic
Date Deposited: 07 May 2014 11:51
Last Modified: 07 May 2014 11:51
URI: http://digre.pmf.unizg.hr/id/eprint/1055

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