The JCV DNA Quantitation Real-Time PCR kit coded JCVDNAQT.CE is intended for the quantitative detection of the JC Virus DNA in human plasma ,urine and cerebrospinal fluid (CSF) with a simultaneous control of the amplification/extraction reaction through an Internal Control (IC). The kit has been adapted for the use on the Real-Time Thermacyclers ABI 7500 Sequence Detection System® (Software SDS version 1.3.1, Applied Biosystems™*) or MX3000P® (Software MxPro version 4.01, Stratagene™***) or CFX96 (Software CFX manager version 1.7, Biorad™**).
NTRODUCTION
Human Polyoma JC Virus (Polyomavirus hominis 2) belongs to the genus Polyomaviridae, which are non enveloped DNA viruses containing circle double-stranded DNAof about 5 KB in size. It was first identified in 1965 by electron microscopy in cases of progressive multifocal leukoencephalopathy (PML) and was first isolated in culture in 1971.
Up to 90% of adults are seropositive. The primary route of transmission has not been firmly defined but an association of primary polyomavirus infection with mild respiratory tract disease and cystitis has been reported. The presence of JC Virus in stool samples from children suggest that a fecal-oral route of transmission may play a role in virus dissemination. After entering the host, JC Virus frequently establish latent and/or persistent infections with persistent presence in different organs, including the kidney. The virus remains quiescent unless a natural or iatrogenic state of immunosoppression is imposed.
RINCIPLE OF THE TEST
The JCVDNAQT.CE Kit is based on a Real Time chemistry which uses specific Primers and Probes.
JCV DNA, recovered from the biological sample under investigation through an extraction step, is 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 JCV unique genomic sequence.
Internal Control (IC) serves as an amplification/extraction control for each individually processed specimen aiming to the identification of reaction inhibitors.
An external standard curve is supplied allowing the determination of the viral load.
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