Detection of reverse transcriptase in culture medium for mammary tumour cell lines: a comparison of an established radio-labelling technique and a contemporary non-isotopic technique.
Classically, radio-label techniques have been employed to analyse biological samples for reverse transcriptase (RT) activity. More recently, however, non-isotopic kits have been developed for retroviral quantification. Nevertheless, until the present investigation it has not been known if these contemporary methods are more sensitive at detecting reverse transcriptase activity. In our study, a non-isotopic ELISA method was shown to be considerably more sensitive than the radio-label technique at detecting reverse transcriptase in growth medium used to culture the murine breast cancer cell line GR/A. Using the ELISA, less reverse transcriptase activity was demonstrated in growth medium from human mammary adenocarcinoma MCF-7 cells than the murine source. This ELISA did not detect reverse transcriptase activity from a pure source of Moloney murine leukaemia virus. In light of this, the broad applicability of this ELISA for reverse transcriptase from different viral sources must be investigated before it can be used to monitor biological supernatants for the presence of retroviruses.
Hughes
G A
GA
Department of Biological and Biomedical Sciences, University of Ulster, Coleraine, Co. Londonderry, Northern Ireland, UK.
Barnett
Y A
YA
Allen
J M
JM
D000818:Animals; D003470:Culture Media; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D051379:Mice; D008979:Moloney murine leukemia virus / Q000201:enzymology; D010761:Phosphorus Radioisotopes; D012194:RNA-Directed DNA Polymerase / Q000302:isolation & purification*; D012190:Retroviridae / Q000201:enzymology*; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
D003470:Culture Media; D010761:Phosphorus Radioisotopes; D012194:RNA-Directed DNA Polymerase