Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Lee, Li Ling, Lin, Lin, Bell, David S., Levine, Susan, Hanson, Maureen R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
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
_version_ 1782233666874769408
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
work_keys_str_mv AT leeliling sensitivityofpcrassaysformurinegammaretrovirusesandmousecontaminationinhumanbloodsamples
AT linlin sensitivityofpcrassaysformurinegammaretrovirusesandmousecontaminationinhumanbloodsamples
AT belldavids sensitivityofpcrassaysformurinegammaretrovirusesandmousecontaminationinhumanbloodsamples
AT levinesusan sensitivityofpcrassaysformurinegammaretrovirusesandmousecontaminationinhumanbloodsamples
AT hansonmaureenr sensitivityofpcrassaysformurinegammaretrovirusesandmousecontaminationinhumanbloodsamples