Hughes G A, Barnett Y A, Allen J M
Department of Biological and Biomedical Sciences, University of Ulster, Coleraine, Co. Londonderry, Northern Ireland, UK.
Br J Biomed Sci. 1995 Mar;52(1):2-8.
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.
传统上,放射性标记技术已被用于分析生物样品中的逆转录酶(RT)活性。然而,最近已经开发出用于逆转录病毒定量的非同位素试剂盒。尽管如此,直到本研究之前,尚不清楚这些现代方法在检测逆转录酶活性方面是否更敏感。在我们的研究中,一种非同位素ELISA方法在检测用于培养小鼠乳腺癌细胞系GR/A的生长培养基中的逆转录酶时,显示出比放射性标记技术灵敏得多。使用ELISA,在人乳腺腺癌MCF-7细胞的生长培养基中显示出的逆转录酶活性低于小鼠来源。该ELISA未检测到来自纯莫洛尼鼠白血病病毒的逆转录酶活性。鉴于此,在将该ELISA用于监测生物上清液中是否存在逆转录病毒之前,必须研究其对不同病毒来源逆转录酶的广泛适用性。