Polyoma Virus JC and BK

Polyomaviruses JC and BK are double stranded DNA viruses which are ubiquitous in the human population. Primary infections are usually asymptomatic. JCV is the causative agent of the neurological disease progressive multifocal leukoencephalopathy, which occurs frequently in AIDS patients. BKV is associated with hemorrhagic cystitis, ureteral stenosis and other urinary tract disease¹ in transplant patients with immunosuppressive therapy.

¹BK virus associated renal cell carcinoma: case presentation with optimized PCR and other diagnostic tests. Narayanan M, Szymanski J, Slavcheva E, Rao A, Kelly A, Jones K, Jaffers G. Am J Transplant 7(6):1666-71 (2007).

kKit with reagents for the quantitative detection of Polyomaviruses JC and BK DNA using the LightCycler^® Instruments 1.x / 2.0 and 480 II.

Description

A 172 bp fragment (Polyomavirus JC) and a 175 bp fragment (Polyomavirus BK) of the Small t-Antigen are amplified with specific primers and detected with probes labeled with LightCycler^® Red 705 (detected in channel 705) for Polyomavirus JC or with LightCycler Red^® 640 (detected in channel 640) for Polyomavirus BK respectively. The PCR products are identified by running melting curves with specific melting points (Tm) of 68°C for Polyomavirus JC in channel 705 and of 64°C for Polyomavirus BK in channel 640.

An additional PCR product of 658 bp is formed from the internal positive control DNA. This control will not interfere with the Polyoma specific reactions. The amplification will usually fail in the presence of higher concentrated Polyoma DNA samples (1,000 - 10,000 copies or higher) but it will display a melting curve signal in negative and low-concentrated samples. The probes are labeled with the dye LC705. Detection is recorded in channel 705; the specific Tm is about 50°C.

Additional reagents and items required

Color Compensation Kit 530/640/690	40-0318-00
LightCycler® FastStart DNA Master HybProbe	03 003 248 001
Capillaries (20 µl) (LC1.x / 2.0 Instruments)	04 929 292 001
LC 480 Multiwell Plate 384, white	04 729 749 001

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