Abstract
Real-time RT-PCR has become the method of choice for automated detection of viral RNA target sequences in the clinical laboratory. Besides commercially available certified test systems, a variety of so-called in-house methods have been described in the literature. Generally, appropriate validation and continuous quality control are mandatory if these in-house-developed assays are used in clinical diagnostics. In this chapter, an in-house HIV-1 real-time RT-PCR assay for blood donor screening is described. The procedure includes the pooling of plasma samples, viral RNA isolation, and subsequent detection of amplification in real-time one-step RT-PCR. The validation considers the specificity, the sensitivity on HIV-1 genomic variants, and the robustness of the assay.
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Acknowledgments
The author would like to thank Miriam Herzig and Regina Maurer for expert technical assistance, as well as Jürgen Bach, Bernd Pötzsch, and Jutta Rox for critical reading of the manuscript.
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Müller, J. (2010). Real-time RT-PCR for Automated Detection of HIV-1 RNA During Blood Donor Screening. In: King, N. (eds) RT-PCR Protocols. Methods in Molecular Biology, vol 630. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-629-0_20
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DOI: https://doi.org/10.1007/978-1-60761-629-0_20
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