Volume 8 Supplement 1
A prototype RT-PCR assay for detection of XMRV in multiple human sample types
© Tang et al; licensee BioMed Central Ltd. 2011
Published: 6 June 2011
Xenotropic murine leukemia virus-related virus (XMRV) has been reported to be associated with prostate cancer and chronic fatigue syndrome. To help resolve the role of XMRV in human disease, it is critical to develop sensitive and accurate PCR assays for XMRV detection.
Materials and methods
The automated m2000 system RT-PCR assays detect the pol and env regions of XMRV in whole blood, plasma, urine, and cell pellets. Assay performance was assessed by testing two blinded panels prepared by the Blood XMRV Scientific Research Working Group (SRWG), as well as clinical specimens.
For the SRWG whole blood panel, all six XMRV negative samples were assay negative, while all the 22Rv1-spiked samples from 0.5 cells/mL to 9,900 cells/mL were detected (3/3 for each panel member). For the SRWG plasma panel, all six XMRV negative samples were assay negative, while the 22Rv1 supernatant-spiked plasma samples were detected as positive, 1/3 for the 3.2 XMRV copies/mL panel member, 2/3 for the 16 copies/mL panel member, and 3/3 for panel members containing > 80 copies/mL. For screening of clinical specimen, assay positive results were obtained for 10% (2/20) of prostate cancer FFPE specimens, 0.5% (2/400) of prostate cancer urine pellets, and 0% (0/135) of the cervical swab specimens.
We developed an automated high throughput real-time RT-PCR assay (with internal control) to detect XMRV. Use of this assay should assist in elucidation of the role of XMRV in human disease.
This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.