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Real-time RT-PCR for automated detection of HIV-1 RNA during blood donor screening.

Authors
  • Müller, Jens
Type
Published Article
Journal
Methods in Molecular Biology
Publication Date
Jan 01, 2010
Volume
630
Pages
319–335
Identifiers
DOI: 10.1007/978-1-60761-629-0_20
PMID: 20301006
Source
Medline
License
Unknown

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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