(RT)-qPCR for detection of and differentiation between RNA and DNA of HIV-1-based lentiviral vectors.

The purpose of the described method is the detection of and differentiation between RNA and DNA of HIV-derived lentiviral vectors (LV) in cell culture supernatants and swab samples. For the analytical surveillance of genetic engineering operations methods for the detection of the HIV-1 based LV generations are required. Furthermore, for research issues, it is important to prove the absence of LV particles for downgrading experimental settings in terms of the biosafety level. Here a qPCR method targeting the LTR U5 subunit and the start sequence of the packaging signal ψ is described. Numerous controls are included in order to monitor the technical procedure.