A ready-to-use system for the isolation and detection of XMRV viral RNA from urine using end-point RT-PCR

  • Rapid isolation of high quality viral RNA from urine
  • Contains two ready-to-use 2X RT-PCR Master Mixes
  • High sensitivity and specificity
  • Includes an isolation control and a PCR control
  • Primer set and controls also available separately
  • Ideal for use in:
    1. Surveillance of Drug Resistant Pathogens
    2. Epidemiologial Studies
    3. Field Surveillance of Pathogens
    4. Surveys

ItemsCat. #Size
Urine-Based XMRV RT-PCR Detection Kit 34700 24 tests

Urine-Based XMRV RT-PCR Detection Kit

Xenotropic murine leukemia virus-related virus (XMRV) belongs to the family Retroviridae and the genus gammaretrovirus. The virus was first described in 2006 and has since been isolated from human biological samples. XMRV has a single-stranded RNA genome that replicates through a DNA intermediate. The virus gets its name due to its close relationship with the murine leukemia viruses (MuLVs). The viral genome is approximately 8100 nucleotides in length and is 95% identical with several endogenous retroviruses of mice. While gammaretroviruses have well-characterized oncogenic effects in animals, they have not been shown to cause human cancers. However, XMRV was recently discovered in human prostate cancers and is the first gammaretrovirus known to infect humans. In addition to prostate cancer, a possible association with chronic fatigue syndrome has been reported, however it has yet to be established whether XMRV is a cause of this disease. The causal role of XMRV in cancer has yet to be established and the virus does not appear to be capable of transforming cells directly.

Norgen’s Urine-Based XMRV RT-PCR Detection Kit is a ready-to-use system for the isolation and detection of XMRV from urine. First the kit contains components for the rapid isolation of total RNA, including viral RNA, from the urine samples using spin-column chromatography based on Norgen’s proprietary resin (Nucleic Acid Isolation). Second the kit contains XMRV RT-PCR Master Mix and controls for PCR Amplification, as well as a Control RT-PCR Master Mix to allow for amplification of both an Isolation Control and a PCR Control. The amplified PCR products are then detected using agarose gel electrohporesis. Alternatively, detection can be performed by real-time PCR using melt curves (Detection). Please see the flowchart to the right.

The XMRV Master Mix contains reagents and enzymes for the specific amplification of a 300 bp region of XMRV. In addition, Norgen’s Urine-Based XMRV RT-PCR Detection Kit contains a second Master Mix, the Control 2x RT-PCrR Master Mix, which can be used to identify possible PCR inhibition and/or inadequate isolation. The kit is designed to allow for the testing of 24 samples. the XMRV RT-PCR Primer Set and Controls are also available separately for end-point RT-PCR detection.

  • Non Invasive, Non Infectious Sample Source – Urine is not infectious for most human pathogens.
  • Isolate High Quality, Inhibitor-Free RNA - Removal of highly concentrated salts, metabolic wastes and proteins provides high quality, concentrated RNA.
  • Rapid Procedure - Isolation and detection of XMRV in less than 3 hours.
  • Accuracy in Interpreting Results – The amplification bands are clearly distinct, and the melt curves do not overlap
  • High Sensitivity and Specificity - The limit of detection is less than 2000 VP/mL of urine
  • Purchase Primer Set and Controls Separately - The XMRV Primer set as well as the positive control and negative control can be purchased separately

Linear Range

• The linear range of Norgen’s XMRV RT-PCR Kit Dx was determined by analyzing a dilution series of XMRV quantitative standard ranging from 8.46 x 109 VP/µl to 1 x 10-1 IU/µl.
• Each dilution has been tested in replicates (n = 4) using Norgen’s XMRV RT-PCR Kit Dx on 1X TAE, 1.7% Agarose gels.
• The linear range of Norgen’s XMRV RT-PCR Kit Dx has been determined to cover concentrations from 9 VP/µl to at least 8 x 106 VP/µl

    Urine-Based XMRV PCR Detection Kit Contents
    RNA Lysis Solution
    RNA Wash Solution
    RNA Elution Solution
    Mini Filter Spin Columns
    Collection Tubes
    Elution tubes (1.7 mL)
    XMRV 2x RT-PCR Master Mix
    Control 2x RT-PCR Master Mix
    Isolation Control (IsoC)
    XMRV Positive Control (PosC)
    Nuclease-Free Water
    Norgen’s DNA Marker
    Product Insert

    XMRV RT-PCR Primer Set and Controls Contents
    XMRV Primer set
    XMRV Positive Control (PosC)
    Nuclease-Free Water

    Storage Conditions and Product Stability

    • The Positive Control (XMRV PosC, red cap) and Isolation Control ( IsoC, orange cap) should be stored at -70°C. If needed, make aliquots of the controls according to the volume used in the protocol (10 µL of XMRV PosC or 15 µL of XMRV IsoC) prior to freezing.
    • The XMRV 2X RT-PCR Mastermix and Control 2X RT-PCR Mastermix should be stored at -20°C. Make appropriate aliquots if needed
    • All other kit components may be stored at room temperature
    • The XMRV 2X RT-PCR Mastermix, Control 2X RT-PCR Mastermix, XMRV Postive Control (PosC) and Isolation Control (IsoC) should not undergo repeated freeze-thaw (a maximum freeze-thaw of three times).
    • Allow reagents to thaw at room temperature prior to use
    • After addition of samples to RT-PCR Master Mix use within one hour

    Title Detection of Xenotropic Murine Leukemia Virus-Related Virus in Prostate Biopsy Samples.
    Journal J Coll Physicians Surg Pak. 2014.
    Authors Faraz Ahmed Baig, Talat Mirza, Rafiq Khanani and Saeed Khan.