uu.seUppsala University Publications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Search for DNA of exogenous mouse mammary tumor virus-related virus in human breast cancer samples
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Clinical Virology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Clinical Virology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Clinical Virology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Genetics and Pathology.
Show others and affiliations
2007 (English)In: Journal of General Virology, ISSN 0022-1317, E-ISSN 1465-2099, Vol. 88, 1806-1809 p.Article in journal (Refereed) Published
Abstract [en]

Earlier reports of a human exogenous retrovirus (HMTV) related closely to mouse mammary tumor virus (MMTV) led us to search for these viral sequences in breast cancer tissues and normal tissues. A real-time PCR was developed based on MMTV and published HMTV envelope sequences. The real-time PCR method can detect one to ten copies of MMTV target DNA. Tissue samples were collected prospectively from 18 breast cancer patients and 11 non-malignant control cases, as well as peripheral blood leukocytes from the same women. Despite the high sensitivity of the real-time PCR method used, none of the samples were positive for HMTV DNA or RNA. The absence of HMTV DNA in both breast cancer samples and controls indicates either that the concentration of putative HMTV DNA in the breast cancers was too low for detection or that it did not exist there.

Place, publisher, year, edition, pages
2007. Vol. 88, 1806-1809 p.
Keyword [en]
Adult, Aged, Aged; 80 and over, Base Sequence, Betaretrovirus/genetics/*isolation & purification, Breast Neoplasms/*virology, DNA; Viral/*analysis/genetics, Female, Humans, Mammary Tumor Virus; Mouse/*genetics, Middle Aged, Molecular Sequence Data, Polymerase Chain Reaction/methods, Prospective Studies, Sequence Alignment
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:uu:diva-11879DOI: 10.1099/vir.0.82767-0ISI: 000247087900020PubMedID: 17485542OAI: oai:DiVA.org:uu-11879DiVA: diva2:39648
Available from: 2007-11-01 Created: 2007-11-01 Last updated: 2017-12-11Bibliographically approved
In thesis
1. Detection and Quantification of Variable Viral RNA by Real-Time PCR Assays
Open this publication in new window or tab >>Detection and Quantification of Variable Viral RNA by Real-Time PCR Assays
2008 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

As the area of nucleic acid based technologies develops, so will our understanding of how structural variations in DNA and RNA pathogens are associated with disease. The overall goal of this thesis is the development of broadly targeted measurement techniques for variable viral RNA by Real-Time PCR (here referred to as quantitative reverse transcriptase PCR, QRT-PCR).

In papers I & II, broadly targeted and specific QRT-PCRs were used to study expression of endogenous and exogenous betaretrovirus sequences in human tissues. Results from human tissues demonstrated endogenous betaretrovirus expression in a tissue-specific manner, highest in reproductive tissues. Despite the high sensitivity, no exogenous betaretrovirus was found in human breast cancer samples. The limits of primer and probe degeneracy for detection of a diverse set of retroviral sequences was evaluated. These methods are useful for further investigations on the pathophysiological contribution(s) of endogenous betaretrovirus and to investigate whether an exogenous betaretrovirus is involved in human breast cancer.

In papers III & IV, we developed and applied broadly targeted one-step QRT-PCRs for influenza viruses and coronaviruses. In addition to the generic primers, two novel probe design strategies were used in order to be able to broadly amplify these diverse sets of viruses: A triplex system for simultaneous detection and quantification of influenza A, B and C (3QRT-PCR and further developed 3QRT-PCR-MegB; where MegB stands for MegaBeacon) based on TaqMan® and MegB probes, and a pan-CoV QRT-PCR, based on three TaqMan® probes i.e., degeneracy was distributed on three probes. Probe fault tolerance was thus increased in two ways, either with short probes with/without locked nucleic acid (LNA) nucleotides concentrated to conserved stretches, or with long probes (MegB), compensating mismatching positions with many matching ones. Clinical samples, negative by antigen detection with immunofluorescence (IFA), were influenza A positive with 3QPCR-MegB. Avian pooled samples, negative with an earlier pan-CoV QPCR, came out positive with the triple-probe system. Assay evaluation with clinical samples and reference strains revealed good clinical diagnostic potential.

Thus, the thesis describes several strategies to counteract sequence variation of RNA viruses and describes a set of broadly targeted QRT-PCRs useful for scientific screening or diagnostics of betaretroviruses and respiratory viruses.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2008. 67 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 365
Keyword
Virology, Diagnosis of Viral RNA, QPCR, RNA virus, broadly targeted PCR, probe design strategy, Infectious diseases
Research subject
Medicine
Identifiers
urn:nbn:se:uu:diva-9193 (URN)978-91-554-7251-1 (ISBN)
Public defence
2008-09-05, Hörsalen, Baktlab ing D1, Dag Hammarskjölds väg 17, Uppsala, 13:00 (English)
Opponent
Supervisors
Available from: 2008-08-15 Created: 2008-08-15 Last updated: 2011-02-28Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textPubMedhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Retrieve&list_uids=17485542&dopt=Citation

Authority records BETA

Muradrasoli, ShamanWärnberg, FredrikBlomberg, Jonas

Search in DiVA

By author/editor
Muradrasoli, ShamanWärnberg, FredrikBlomberg, Jonas
By organisation
Clinical VirologyDepartment of Genetics and PathologyEndocrine Surgery
In the same journal
Journal of General Virology
Medical and Health Sciences

Search outside of DiVA

GoogleGoogle Scholar

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 436 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf