Voisset C, Tönjes R R, Breyton P, Mandrand B, Paranhos-Baccalà G
Unité Mixte de Recherche 2142 CNRS-bioMérieux, Ecole Normale Supérieure de Lyon, 46, allée d'Italie, 69364 Cedex 07, Lyon, France.
J Virol Methods. 2001 May;94(1-2):187-93. doi: 10.1016/s0166-0934(01)00255-5.
The polymerase enhanced reverse transcriptase (PERT) assay is a highly sensitive assay for the detection of reverse transcriptase (RT) activity in culture supernatants of retrovirus-producing cells. However, some cellular DNA-dependent DNA polymerases exhibit RT-like activities in this assay. A synthetic DNA competitor which suppresses the RT-like activities of cellular DNA-dependent DNA polymerases was used in a modified PERT assay technique for specific detection of RT activity in culture supernatants of retrovirus-producing cells. We determined the optimum condition of the assay and evaluated its specificity. This improved PERT assay is easy to perform and is able to detect minute amounts of purified RT, as well as RT in crude cell lysates and concentrated culture supernatants.
聚合酶增强逆转录酶（PERT）测定法是一种用于检测逆转录病毒产生细胞培养上清液中逆转录酶（RT）活性的高灵敏度测定法。然而，一些细胞DNA依赖性DNA聚合酶在此测定法中表现出类似RT的活性。一种抑制细胞DNA依赖性DNA聚合酶类似RT活性的合成DNA竞争剂被用于一种改良的PERT测定技术中，以特异性检测逆转录病毒产生细胞培养上清液中的RT活性。我们确定了该测定法的最佳条件并评估了其特异性。这种改进的PERT测定法易于操作，能够检测微量的纯化RT，以及粗细胞裂解物和浓缩培养上清液中的RT。
