Principle

Isolation of viral RNA and viral DNA from human plasma or serum samples – for in-vitro diagnostic purposes

The NucleoSpin® Dx Virus kit is a generic system for the isolation and purification of viral nucleic acids from human serum or plasma samples for subsequent in-vitro diagnostic purposes. The kit can be used with fresh and frozen human serum and plasma, stabilized with either EDTA or citrate from common blood collection systems. The kit is designed to be used with any downstream application employing enzymatic amplification and detection of RNA and DNA (e.g., RT-PCR, PCR). The viral nucleic acids isolated and purified with NucleoSpin® Dx Virus can be used in qualitative applications (e.g., RT-PCR or PCR for blood screening) as well as in quantitative applications (e.g., detection of viral load by qPCR) employing diagnostic nucleic acid amplification techniques.

NucleoSpin Dx Virus Procedure

NucleoSpin® Dx Virus is based on proven NucleoSpin® silica-membrane technology and provides an easy way to isolate viral RNA and viral DNA from 150 μl of serum or plasma samples. Purified RNA and DNA are ready to use for downstream amplifications like RT-PCR or PCR. The NucleoSpin® Dx Virus procedure is based on a series of simple steps: First, the serum or plasma samples are lysed in the presence of chaotropic salts. For the purification of viral DNA, Proteinase K is added to the lysis reaction. Lysis buffer and ethanol create appropriate conditions for binding of nucleic acids to the silica membrane of the NucleoSpin® Dx Virus Columns while Carrier RNA improves binding and recovery of low-concentrated viral RNA and DNA. Contaminations (potential PCR inhibitors) like salts, metabolites, and soluble macromolecular cellular components are removed in three washing steps. The nucleic acids are finally eluted in 50 μl low salt buffer or DNase-free water.

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