Lisa Becherer,
University of Freiburg
Germany
Title: Simultaneous detection of HIV and HTLV by mediator displacement loop-mediated isothermal amplification
Biography
Biography: Lisa Becherer,
Abstract
Nucleic acid amplification tests (NAAT) are not only a powerful tool for early diagnosis of HIV infections, NAAT are also a reliable method for HIV viral load measurements during the monitoring of antiretroviral therapy. Additionally, NAAT allow simultaneous (multiplex) detection of different targets enabling the detection of HIV/HTLV co infections. Loopmediated isothermal amplification (LAMP) [1, 2] emerges as a convenient alternative to polymerase chain reaction (PCR) for rapid amplification of target DNA and RNA. As an isothermal NAAT, LAMP does not require expensive equipment for thermo cycling and is therefore especially suitable for point-of-care testing [3]. However, available multiplex detection techniques for LAMP suffer from elaborate assay design as well as time-consuming optimization work. Here we present the first multiplex reverse transcription (RT) LAMP for identification of HIV/HTLV co-infections [4]. The quantitative real-time assay is based on universal mediator and reporter molecules [5] generating a fluorescence signal in the presence of target sequences. During amplification of target DNA the mediator is displaced (step 1, Figure 1). The released mediator hybridizes to the reporter generating a fluorescence signal (step 2, Figure 1) which can be detected.