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
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Urine-Based XMRV RT-PCR Detection Kit - Short Protocol
PDF version
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PRODUCT INFORMATION SHEETS
Urine-Based XMRV RT-PCR Detection Kit - Product Information Sheet
PDF version
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APPLICATION NOTES
POSTERS
MSDS
SUPPLEMENTARY PROTOCOLS
REFERENCES
FREQUENTLY ASKED QUESTIONS
Urine-Based XMRV RT-PCR Detection Kit - Frequently Asked Questions
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Phone: (905) 227-8848 | Toll Free: 1-866-NORGENB (667-4362) | Fax: (905) 227-1061 | email:
info@norgenbiotek.com