The HHV8 DNA Quantitation (QT) Real-Time PCR kit coded HHV8DNAQT.CE is intended for the quantitative detection of Human herpesvirus 8 DNA in human plasma/blood with a simultaneous control of the extraction/amplification reaction through an Internal Control (IC).
The kit has been adapted for the use on the Real-Time Thermacyclers and ABI 7500 Sequence Detection System® (Software SDS version 1.3.1, Applied Biosystems™*) CFX96 ® (Software CFX manager version 1.7 , Biorad™**) or MX3000P (Software MxPro version 4.01, Stratagene™***).
NTRODUCTION
HHV-8 is a member of the Gammaherpesviririnae subfamily. The prevalence of HHV-8 infection varies widely, from approximately 1–3% of the general population in North America to more than 70% in several regions of Africa where HHV-8 is endemic. HHV-8, also known as Kaposi’s sarcoma associated herpesvirus, has been associated with Kaposi’s disease, multicentric Castleman’s disease, primary effusion lymphoma and other plasmablastic lymphoma classified as diffuse large B-cell lymphoma.
The complete HHV-8 genome sequence shows that it has sequence similarities to other gammaherpesviruses including, herpesvirus saimiri (HVS), murine gammaherpesvirus 68 (MHV68) and Epstein-Barr Virus (HHV-4). The ~165 kb genome contains over 80 open reading frames arranged in a long unique region flanked by multiple 801bp terminal repeat units of high G+C content. The long unique region contains blocks of conserved genes found in most herpesviruses, interspersed with blocks of non-homologous genes that are specific for HHV-8 and related viruses.
RINCIPLE OF THE TEST
The HHV8DNAQT.CE Kit is based on a Real Time chemistry which uses specific Primers and Probes.
HHV8 DNA, recovered from the biological sample under investigation through an extraction step, and amplified using Real Time amplification system. The amplified product, is detected and quantified, against the standard curve using a fluorescent reporter dye probe specific for a HHV8 unique genomic sequence.
Heterologous Internal Control (IC) serves as an extraction/amplification control for each individually processed specimen aiming to the identification of reaction inhibitors.
A standard curve is supplied allowing the determination of the viral load.
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