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Sensitivity of PCR Assays for Murine Gammaretroviruses and Mouse Contamination in Human Blood Samples
Gammaretroviruses related to murine leukemia virus (MLV) have variously been reported to be present or absent in blood from chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) patients and healthy controls. Using subjects from New York State, we have investigated by PCR methods whether MLV-r...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3357399/ https://www.ncbi.nlm.nih.gov/pubmed/22629404 http://dx.doi.org/10.1371/journal.pone.0037482 |
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author | Lee, Li Ling Lin, Lin Bell, David S. Levine, Susan Hanson, Maureen R. |
author_facet | Lee, Li Ling Lin, Lin Bell, David S. Levine, Susan Hanson, Maureen R. |
author_sort | Lee, Li Ling |
collection | PubMed |
description | Gammaretroviruses related to murine leukemia virus (MLV) have variously been reported to be present or absent in blood from chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) patients and healthy controls. Using subjects from New York State, we have investigated by PCR methods whether MLV-related sequences can be identified in nucleic acids isolated from whole blood or from peripheral blood mononuclear cells (PBMCs) or following PBMC culture. We have also passaged the prostate cancer cell line LNCaP following incubation with plasma from patients and controls and assayed nucleic acids for viral sequences. We have used 15 sets of primers that can effectively amplify conserved regions of murine endogenous and exogenous retrovirus sequences. We demonstrate that our PCR assays for MLV-related gag sequences and for mouse DNA contamination are extremely sensitive. While we have identified MLV-like gag sequences following PCR on human DNA preparations, we are unable to conclude that these sequences originated in the blood samples. |
format | Online Article Text |
id | pubmed-3357399 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-33573992012-05-24 Sensitivity of PCR Assays for Murine Gammaretroviruses and Mouse Contamination in Human Blood Samples Lee, Li Ling Lin, Lin Bell, David S. Levine, Susan Hanson, Maureen R. PLoS One Research Article Gammaretroviruses related to murine leukemia virus (MLV) have variously been reported to be present or absent in blood from chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) patients and healthy controls. Using subjects from New York State, we have investigated by PCR methods whether MLV-related sequences can be identified in nucleic acids isolated from whole blood or from peripheral blood mononuclear cells (PBMCs) or following PBMC culture. We have also passaged the prostate cancer cell line LNCaP following incubation with plasma from patients and controls and assayed nucleic acids for viral sequences. We have used 15 sets of primers that can effectively amplify conserved regions of murine endogenous and exogenous retrovirus sequences. We demonstrate that our PCR assays for MLV-related gag sequences and for mouse DNA contamination are extremely sensitive. While we have identified MLV-like gag sequences following PCR on human DNA preparations, we are unable to conclude that these sequences originated in the blood samples. Public Library of Science 2012-05-21 /pmc/articles/PMC3357399/ /pubmed/22629404 http://dx.doi.org/10.1371/journal.pone.0037482 Text en Lee et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Lee, Li Ling Lin, Lin Bell, David S. Levine, Susan Hanson, Maureen R. Sensitivity of PCR Assays for Murine Gammaretroviruses and Mouse Contamination in Human Blood Samples |
title | Sensitivity of PCR Assays for Murine Gammaretroviruses and Mouse Contamination in Human Blood Samples |
title_full | Sensitivity of PCR Assays for Murine Gammaretroviruses and Mouse Contamination in Human Blood Samples |
title_fullStr | Sensitivity of PCR Assays for Murine Gammaretroviruses and Mouse Contamination in Human Blood Samples |
title_full_unstemmed | Sensitivity of PCR Assays for Murine Gammaretroviruses and Mouse Contamination in Human Blood Samples |
title_short | Sensitivity of PCR Assays for Murine Gammaretroviruses and Mouse Contamination in Human Blood Samples |
title_sort | sensitivity of pcr assays for murine gammaretroviruses and mouse contamination in human blood samples |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3357399/ https://www.ncbi.nlm.nih.gov/pubmed/22629404 http://dx.doi.org/10.1371/journal.pone.0037482 |
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