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XMRV - testing urine

Discussion in 'XMRV Testing, Treatment and Transmission' started by glenp, Jun 13, 2010.

  1. glenp

    glenp "and this too shall pass"

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    Vancouver Canada suburbs
  2. acer2000

    acer2000 Senior Member

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    Very interesting... how did you find that? Also, is there any data in research that shows that XMRV or MLV can be detected reliably in urine?
     
  3. glenp

    glenp "and this too shall pass"

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    IDK I have NO idea. This boggles me. Looks like a reputable lab??
     
  4. abc123

    abc123

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    Thanks for posting this. I like this part:

    "Ideal for use in:
    1. Surveillance of Drug Resistant Pathogens
    2. Epidemiologial Studies
    3. Field Surveillance of Pathogens
    4. Surveys"

    Who is doing any of those things...?

    I'm all for a good urine test over a blood test, but neat, new tests don't mean anything because at this point 1) the connection between XMRV and disease has not been made, and 2) doctors won't offer any treatments anyway. It's still "all in your head."
     
  5. thegodofpleasure

    thegodofpleasure Player in a Greek Tragedy

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    Matlock, Derbyshire, Uk
    If this is genuine and the test performs in the ways that they claim, then it is a most welcome development.

    However, where have they been in the debate to date?
    Where is the verification that this test works - confirmatory study or the like ?
    Where was the fanfare when the test was announced / perfected ?

    I therefore remain sceptical until the test is verified

    TGOP
     
  6. garcia

    garcia Aristocrat Extraordinaire

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    London, UK
    This is a pcr test, so I'm guessing it will be as useless as the other pcr tests developed so far (other than the WPI/VIP one). No indication that it is any different from those.
     
  7. omerbasket

    omerbasket Senior Member

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    I guess that theoretically PCR of the urine might be more accurate than of the blood. But this all seems to me like another test that is not proven and would probably produce a lot of false negatives or might even find close to zero positives.
     
  8. Impish

    Impish Senior Member

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    Well the German's found XMRV in throat swabs so who knows. The obvious way to test whether or not it actually works is to see if it can detect XMRV in someone who has been confirmed to have it by other means. I don't see any mention of that anywhere... I am guessing that they are thinking that if it is present in prostate cancer XMRV should be detected in urine?
     
  9. Esther12

    Esther12 Senior Member

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    I think it's best to stay sceptical at this point.

    Given the confused state of XMRV research at the moment, it would be a prime target for disreputable testing imo. The difficulties of accurately testing for Lyme has led to a rise of dodgy tests, we could see the same thing with XMRV.
     
  10. VillageLife

    VillageLife Senior Member

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    would be interesting to hear the WPI's opinion on this new test!
     
  11. Min

    Min Senior Member

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    I can't find a price on this - does anyone know what they are charging please?
     
  12. George

    George waitin' fer rabbits

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    South Texas
    I LOVE GLENP (grin)

    But I don't like this test. (growlies) It reminds me too much of Cooperative Diagnostics (who really tried) and Clongen (who really hasn't). A Test without testing the test. You know?

    Great find Glen (sluuuuuuuurrrp)
     
  13. ixchelkali

    ixchelkali Senior Member

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    Long Beach, CA
    Oh, for pity's sake. This one really brings out the skeptic in me. There have been no published studies indicating that XMRV can be found in urine. I'm filing this one under "Preying on sick people," unless and until they show some evidence that it works. Something better than the "Trust us, it works" (dazzel 'em with big words) that they have on their website.

    Cheap! Fast! Get your cut-rate XMRV testing here, folks!

    Please be clear that my distain is aimed at the lab, not at this thread. I appreciate GlenP's posting this.
     
  14. glenp

    glenp "and this too shall pass"

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    I emailed them for more info (not that I am getting the test) They have a toll free # for canadian and us people but am not up to much telephone converstion - if anyone is it would be interesting what they have to say. I will post when or if I get an email from them.

    glen
     
  15. George

    George waitin' fer rabbits

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    Dang, glen the direct approach, why do I never think of that! Smart!
     
  16. usedtobeperkytina

    usedtobeperkytina Senior Member

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    Clay, Alabama
    well, at least someone is interested in marketing a product to us, besides supplements.

    Surely some big labs got to see that we are a gold mine.

    Can't imagine doing my own test. But notice this is for studies also.

    Hey, maybe we just need to do our own research.

    (I am joking folks)

    Tina
     
  17. staj

    staj Guest

    Dear Geln,
    This test is for research use only. As of now we have not validated the test for human diagnostics. The test ought to work on blood, saliva and urine, provided the virus is present in such specimen. Note we have no evidence that XMRV is present in urine. If XMRV is present in urine our kit ought to detect it.
    Note this kit is available only to scientists conducting research and is not for sale to individuals. When the test is validated for human we will let you know:

    Ideal for use in:
    1. Surveillance of Drug Resistant Pathogens
    2. Epidemiologial Studies
    3. Field Surveillance of Pathogens
    4. Surveys
     
  18. omerbasket

    omerbasket Senior Member

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    I wonder: If there is one virus (without any more copies of it - just one particle) in the whole urine sample - would the test detect it? Because you said: "If XMRV is present in urine our kit ought to detect it".
     
  19. glenp

    glenp "and this too shall pass"

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    Staj - are you willing to release the test to other labs in kits of 24 tests?
    Would you release the test to BCCDC or Bio - Labs?

    It looked to me like the test was available for any lab or individual ordering it??

    There is no mention on the site that it is for research only??
    http://norgenbiotek.com/product_res...v_rtpcr_detection_kit__protocol_34700_886.pdf



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    I copied this from the site -- no mention of research??? Where does it say it is for research only???


    info@norgenbiotek.com

    Norgen Biotek Corp.
    3430 Schmon Parkway, Thorold
    Ontario, Canada, L2V 4Y6

    Tel: (905) 227-8848
    Fax: (905) 227-1061
    Urine-Based XMRV RT-PCR Detection Kit (Cat# 34700 )
    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 a ready-to-use 2X RT-PCR Master Mix
    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

    Product Description
    Product Description
    Applications & Data
    Applications & Data
    Specifications
    Specifications Resources
    Resources
    Ordering
    Ordering
    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. 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 heterologous amplification system 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.

    FEATURES AND BENEFITS

    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 Master Mix (with primers and PCR control), as well as the positive control and negative control can be purchased separately




    APPLICATIONS

    A. Isolate High Quality RNA from Urine


    Figure 1. Isolation and Detection of Total RNA from 10 mL Urine Samples with no PCR Inhibition. Total RNA was isolated from two different 10 mL urine samples using Norgen’s Urine Total RNA Purification Maxi Kit (Slurry Format), which uses the same RNA isolation technology as the Urine-Based XMRV RT-PCR Detection Kit. The purified total RNA was then used as the template in an RT-qPCR reaction to detect the human miR-21 gene (Red line) and the human 5S gene (Blue line). Five microlitres of the isolated RNA was used as the template in the RT step, and 5 μL from the RT step was used in the qPCR reaction. As it can be seen, the qPCR was able to successfully detect and amplify both the 5S gene and the the miR-21 gene in all cases, indicating the high quality of the isolated urine total RNA. The black line in the graph above corresponds to the No Template Control.



    B. Outstanding Sensitivity


    Figure 2. Sensitivity of Detection using the Urine-Based XMRV RT-PCR Detection Kit. A representative 1X TAE, 1.7% agarose gel showing the amplification of XMRV at different concentrations (Target). The size of the XMRV target amplicon corresponds to the 300bp band represented by the provided DNA Marker (M). The size of the XMRV Isolation Control (IsoC) corresponds to the 500bp band represented by the provided DNA Marker (M). The XMRV 2X RT-PCR Master Mix contains an XMRV PCR Control (PCRC). The XMRV PCRC controls for PCR inhibition. The size of the XMRV PCRC corresponds to the 150bp band represented by the provided DNA Marker (M). Lanes A-G represents samples spiked with different XMRV concentrations isolated from 10mL urine samples (interpreted as positive results). The XMRV spiked in urine samples is an in vitro transcribed XMRV RNA fragments.



    C. High Specificity for Detection


    Figure 3. Specificity of the Primers Used to Detect XMRV. A representative 1X TAE, 1.7% agarose gel showing the specificity of the primers used to detect XMRV. The size of the XMRV target amplicon corresponds to the 300bp band represented by the provided DNA Marker (M). The specificity of Norgen’s Urine-Based XMRV RT-PCR Detection Kit is first and foremost ensured by the selection of the XMRV specific primers, as well as the selection of stringent reaction conditions. The primers were checked for possible homologies in GenBank published sequences by sequence comparison analyses. Furthermore, the specificity of the XMRV specific primers were tested against most of the known urine-based detected pathogens. Lane A corresponds represents the viral HIV RNA. Lane B represents the viral XMRV RNA, Lane C represents the viral HBV DNA. Lane D represents Malaria DNA. Lane E represents the High Risk HPV viral DNA (HPV 16). Lane F represents the Low Risk HPV viral DNA (HPV 6). Lane G represents the HSV1 viral DNA (HPV 16). Lane H represents the HSV-2 viral DNA.










    Linear Range


    The linear range of Norgen’s Urine-Based XMRV RT-PCR Detection Kit was determined by analyzing a dilution series of XMRV quantitative standard ranging from 8.46 x 109 VP/l to 1 x 10-1 IU/l.
    Each dilution has been tested in replicates (n = 4) using Norgen’s Urine-Based XMRV RT-PCR Detection Kit on 1X TAE, 1.7% Agarose gels.
    The linear range of Norgen’s Urine-Based XMRV RT-PCR Detection Kit has been determined to cover concentrations from 20 VP/l to at least 8 x 106 VP/l
    Under the conditions of Norgen’s Urine RNA Isolation procedure, Norgen’s Urine-Based XMRV RT-PCR detection Kit covers a linear range from 2000 VP/mL urine to at least 8 x 109 VP/mL urine.
    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
    XMRV Isolation Control (IsoC)
    XMRV Positive Control (PosC)
    XMRV Negative Control (NegC)
    Norgen’s DNA Marker
    Product Insert

    XMRV RT-PCR Primer Set and Controls Contents
    XMRV 2x RT-PCR Master Mix (with Primers and PCR Control)
    XMRV Positive Control (PosC)
    XMRV Negative Control (NegC)

    Storage Conditions and Product Stability

    The Positive Control (XMRV PosC, red cap) and Isolation Control (XMRV IsoC, orange cap) should be stored at -70C. If needed, make aliquots of the controls according to the volume used in the protocol (10 L of XMRV PosC or 15 L of XMRV IsoC) prior to freezing.
    The XMRV 2X RT-PCR Mastermix should be stored at -20C. Make appropriate aliquots if needed
    All other kit components may be stored at room temperature
    The XMRV 2X RT-PCR Mastermix, XMRV Postive Control (PosC) and XMRV 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


    PROTOCOL
    Urine-Based XMRV RT-PCR Detection Kit - Protocol
    PDF version
    --------------------------------------------------------------------------------

    Urine-Based XMRV RT-PCR Detection Kit - Short Protocol
    PDF version
    --------------------------------------------------------------------------------


    PRODUCT INFORMATION SHEETS
    Urine-Based XMRV RT-PCR Detection Kit - Product Information Sheet
    PDF version
    --------------------------------------------------------------------------------


    APPLICATION NOTES

    POSTERS


    MSDS

    SUPPLEMENTARY PROTOCOLS

    REFERENCES

    FREQUENTLY ASKED QUESTIONS
    Urine-Based XMRV RT-PCR Detection Kit - Frequently Asked Questions

    --------------------------------------------------------------------------------


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  20. garcia

    garcia Aristocrat Extraordinaire

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    London, UK
    I'd imagine the WPI would say that these people were taking the p***. :D
     

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